US20230172975A1 - Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same - Google Patents
Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same Download PDFInfo
- Publication number
- US20230172975A1 US20230172975A1 US17/708,253 US202217708253A US2023172975A1 US 20230172975 A1 US20230172975 A1 US 20230172975A1 US 202217708253 A US202217708253 A US 202217708253A US 2023172975 A1 US2023172975 A1 US 2023172975A1
- Authority
- US
- United States
- Prior art keywords
- electrode
- suspension
- nanocrystals
- gold
- metallic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 239000002159 nanocrystal Substances 0.000 title claims abstract description 351
- 239000000725 suspension Substances 0.000 title claims abstract description 259
- JUWSSMXCCAMYGX-UHFFFAOYSA-N gold platinum Chemical compound [Pt].[Au] JUWSSMXCCAMYGX-UHFFFAOYSA-N 0.000 title claims abstract description 17
- 238000004519 manufacturing process Methods 0.000 title abstract description 51
- 239000000463 material Substances 0.000 claims abstract description 86
- 229910052751 metal Inorganic materials 0.000 claims abstract description 75
- 239000002184 metal Substances 0.000 claims abstract description 75
- 239000000126 substance Substances 0.000 claims abstract description 38
- -1 chlorine ions Chemical class 0.000 claims abstract description 30
- 239000000460 chlorine Substances 0.000 claims abstract description 19
- 229910052801 chlorine Inorganic materials 0.000 claims abstract description 19
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims abstract description 16
- 150000001805 chlorine compounds Chemical class 0.000 claims abstract description 9
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 389
- 239000010931 gold Substances 0.000 claims description 187
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 177
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 169
- 238000012545 processing Methods 0.000 claims description 151
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 138
- 229910052697 platinum Inorganic materials 0.000 claims description 137
- 229910052737 gold Inorganic materials 0.000 claims description 129
- 239000003623 enhancer Substances 0.000 claims description 127
- 239000000203 mixture Substances 0.000 claims description 119
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 88
- 239000000470 constituent Substances 0.000 claims description 85
- 239000002245 particle Substances 0.000 claims description 61
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 29
- 230000003197 catalytic effect Effects 0.000 claims description 14
- 230000009089 cytolysis Effects 0.000 claims description 12
- 239000007900 aqueous suspension Substances 0.000 claims description 6
- 229910001260 Pt alloy Inorganic materials 0.000 claims description 4
- 229910001020 Au alloy Inorganic materials 0.000 claims description 2
- 239000007971 pharmaceutical suspension Substances 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 298
- 230000008569 process Effects 0.000 abstract description 154
- 239000002105 nanoparticle Substances 0.000 abstract description 153
- 238000011282 treatment Methods 0.000 abstract description 77
- 239000000084 colloidal system Substances 0.000 abstract description 53
- 239000003638 chemical reducing agent Substances 0.000 abstract description 52
- 230000015572 biosynthetic process Effects 0.000 abstract description 32
- 239000002994 raw material Substances 0.000 abstract description 29
- 238000006722 reduction reaction Methods 0.000 abstract description 27
- 239000004094 surface-active agent Substances 0.000 abstract description 22
- 229910021645 metal ion Inorganic materials 0.000 abstract description 19
- 239000012535 impurity Substances 0.000 abstract description 14
- 239000003381 stabilizer Substances 0.000 abstract description 14
- 239000002082 metal nanoparticle Substances 0.000 abstract description 4
- 201000010099 disease Diseases 0.000 abstract description 3
- 230000006806 disease prevention Effects 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 239000008194 pharmaceutical composition Substances 0.000 abstract description 2
- 238000002560 therapeutic procedure Methods 0.000 abstract description 2
- 239000007788 liquid Substances 0.000 description 220
- 210000004027 cell Anatomy 0.000 description 210
- 241000699670 Mus sp. Species 0.000 description 163
- 206010028980 Neoplasm Diseases 0.000 description 142
- 210000002381 plasma Anatomy 0.000 description 118
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 115
- 241001465754 Metazoa Species 0.000 description 102
- 239000000243 solution Substances 0.000 description 85
- 239000000523 sample Substances 0.000 description 84
- 241000699666 Mus <mouse, genus> Species 0.000 description 76
- 150000001875 compounds Chemical class 0.000 description 69
- 241000894007 species Species 0.000 description 69
- 150000002500 ions Chemical class 0.000 description 64
- 239000013078 crystal Substances 0.000 description 61
- 230000006870 function Effects 0.000 description 61
- 150000002739 metals Chemical class 0.000 description 47
- 238000004627 transmission electron microscopy Methods 0.000 description 46
- 230000012010 growth Effects 0.000 description 45
- 238000012360 testing method Methods 0.000 description 41
- 235000011118 potassium hydroxide Nutrition 0.000 description 38
- 230000000694 effects Effects 0.000 description 36
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 34
- 238000000502 dialysis Methods 0.000 description 34
- 108010088751 Albumins Proteins 0.000 description 30
- 102000009027 Albumins Human genes 0.000 description 30
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 30
- 239000002953 phosphate buffered saline Substances 0.000 description 30
- 229960001814 trypan blue Drugs 0.000 description 30
- 238000006243 chemical reaction Methods 0.000 description 29
- 239000003651 drinking water Substances 0.000 description 29
- 235000020188 drinking water Nutrition 0.000 description 29
- 238000002360 preparation method Methods 0.000 description 29
- 210000004881 tumor cell Anatomy 0.000 description 28
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 27
- 229960004316 cisplatin Drugs 0.000 description 27
- 239000001963 growth medium Substances 0.000 description 27
- 239000000047 product Substances 0.000 description 27
- 201000011510 cancer Diseases 0.000 description 26
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 24
- 238000013459 approach Methods 0.000 description 24
- 238000001479 atomic absorption spectroscopy Methods 0.000 description 24
- 238000009739 binding Methods 0.000 description 24
- 230000010261 cell growth Effects 0.000 description 24
- 238000013461 design Methods 0.000 description 24
- 238000009826 distribution Methods 0.000 description 24
- 230000004614 tumor growth Effects 0.000 description 24
- 238000000576 coating method Methods 0.000 description 23
- 238000005755 formation reaction Methods 0.000 description 23
- 239000012528 membrane Substances 0.000 description 23
- 230000036541 health Effects 0.000 description 21
- 239000000956 alloy Substances 0.000 description 20
- 229910045601 alloy Inorganic materials 0.000 description 20
- 230000027455 binding Effects 0.000 description 20
- 239000007924 injection Substances 0.000 description 20
- 229940090044 injection Drugs 0.000 description 20
- 238000002347 injection Methods 0.000 description 20
- 210000004379 membrane Anatomy 0.000 description 20
- 238000009472 formulation Methods 0.000 description 19
- 239000012530 fluid Substances 0.000 description 18
- 239000012634 fragment Substances 0.000 description 18
- 238000011081 inoculation Methods 0.000 description 18
- 238000000926 separation method Methods 0.000 description 18
- 238000004458 analytical method Methods 0.000 description 17
- 230000001093 anti-cancer Effects 0.000 description 17
- 239000013642 negative control Substances 0.000 description 17
- 238000010899 nucleation Methods 0.000 description 17
- 239000013641 positive control Substances 0.000 description 17
- 238000002296 dynamic light scattering Methods 0.000 description 16
- 230000006698 induction Effects 0.000 description 16
- 230000006911 nucleation Effects 0.000 description 16
- 230000009467 reduction Effects 0.000 description 16
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 15
- 230000003993 interaction Effects 0.000 description 15
- 239000012071 phase Substances 0.000 description 15
- 241000282414 Homo sapiens Species 0.000 description 14
- 230000002411 adverse Effects 0.000 description 14
- 239000011248 coating agent Substances 0.000 description 14
- 238000005259 measurement Methods 0.000 description 14
- 229920003023 plastic Polymers 0.000 description 14
- 239000004033 plastic Substances 0.000 description 14
- 150000003839 salts Chemical class 0.000 description 14
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 13
- 238000010923 batch production Methods 0.000 description 13
- 230000001143 conditioned effect Effects 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 13
- 239000010408 film Substances 0.000 description 13
- 239000007789 gas Substances 0.000 description 13
- 239000000356 contaminant Substances 0.000 description 12
- 235000005911 diet Nutrition 0.000 description 12
- 230000037213 diet Effects 0.000 description 12
- 239000011521 glass Substances 0.000 description 12
- 239000002609 medium Substances 0.000 description 12
- 239000008188 pellet Substances 0.000 description 12
- 239000011850 water-based material Substances 0.000 description 12
- 230000002829 reductive effect Effects 0.000 description 11
- 235000011121 sodium hydroxide Nutrition 0.000 description 11
- 230000006907 apoptotic process Effects 0.000 description 10
- 238000009295 crossflow filtration Methods 0.000 description 10
- 239000010410 layer Substances 0.000 description 10
- 230000000394 mitotic effect Effects 0.000 description 10
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 10
- 108090000623 proteins and genes Proteins 0.000 description 10
- 239000002202 Polyethylene glycol Substances 0.000 description 9
- 230000008859 change Effects 0.000 description 9
- 239000002131 composite material Substances 0.000 description 9
- 239000008367 deionised water Substances 0.000 description 9
- 230000005684 electric field Effects 0.000 description 9
- 238000000724 energy-dispersive X-ray spectrum Methods 0.000 description 9
- 239000007928 intraperitoneal injection Substances 0.000 description 9
- 229920001223 polyethylene glycol Polymers 0.000 description 9
- 229920000642 polymer Polymers 0.000 description 9
- 239000001488 sodium phosphate Substances 0.000 description 9
- 238000003786 synthesis reaction Methods 0.000 description 9
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 8
- 238000000862 absorption spectrum Methods 0.000 description 8
- 230000000712 assembly Effects 0.000 description 8
- 238000000429 assembly Methods 0.000 description 8
- 125000004429 atom Chemical group 0.000 description 8
- 210000000481 breast Anatomy 0.000 description 8
- 210000001072 colon Anatomy 0.000 description 8
- 230000035622 drinking Effects 0.000 description 8
- 238000000338 in vitro Methods 0.000 description 8
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 8
- 230000033001 locomotion Effects 0.000 description 8
- 239000004417 polycarbonate Substances 0.000 description 8
- 239000011734 sodium Substances 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- 231100000419 toxicity Toxicity 0.000 description 8
- 230000001988 toxicity Effects 0.000 description 8
- 230000004580 weight loss Effects 0.000 description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 7
- 230000004568 DNA-binding Effects 0.000 description 7
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- 239000002253 acid Substances 0.000 description 7
- 239000012298 atmosphere Substances 0.000 description 7
- 238000004630 atomic force microscopy Methods 0.000 description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 239000012148 binding buffer Substances 0.000 description 7
- 229910052799 carbon Inorganic materials 0.000 description 7
- 238000001311 chemical methods and process Methods 0.000 description 7
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 7
- 238000011156 evaluation Methods 0.000 description 7
- 230000003862 health status Effects 0.000 description 7
- 239000002086 nanomaterial Substances 0.000 description 7
- 229910052760 oxygen Inorganic materials 0.000 description 7
- 239000001301 oxygen Substances 0.000 description 7
- 239000012465 retentate Substances 0.000 description 7
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 7
- 229910000406 trisodium phosphate Inorganic materials 0.000 description 7
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 7
- 241001354243 Corona Species 0.000 description 6
- 206010015548 Euthanasia Diseases 0.000 description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 6
- 108010020147 Protein Corona Proteins 0.000 description 6
- 230000015556 catabolic process Effects 0.000 description 6
- 238000006555 catalytic reaction Methods 0.000 description 6
- 230000001276 controlling effect Effects 0.000 description 6
- 230000034994 death Effects 0.000 description 6
- 230000007423 decrease Effects 0.000 description 6
- 230000002950 deficient Effects 0.000 description 6
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 6
- 238000002695 general anesthesia Methods 0.000 description 6
- 238000003306 harvesting Methods 0.000 description 6
- 238000002173 high-resolution transmission electron microscopy Methods 0.000 description 6
- 238000001727 in vivo Methods 0.000 description 6
- 238000003780 insertion Methods 0.000 description 6
- 230000037431 insertion Effects 0.000 description 6
- 244000052769 pathogen Species 0.000 description 6
- 230000001717 pathogenic effect Effects 0.000 description 6
- 235000021401 pellet diet Nutrition 0.000 description 6
- 239000011736 potassium bicarbonate Substances 0.000 description 6
- 229910000028 potassium bicarbonate Inorganic materials 0.000 description 6
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 6
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 6
- 210000002307 prostate Anatomy 0.000 description 6
- 238000000159 protein binding assay Methods 0.000 description 6
- 230000005855 radiation Effects 0.000 description 6
- 230000004044 response Effects 0.000 description 6
- 238000001223 reverse osmosis Methods 0.000 description 6
- 238000001350 scanning transmission electron microscopy Methods 0.000 description 6
- JHJLBTNAGRQEKS-UHFFFAOYSA-M sodium bromide Chemical compound [Na+].[Br-] JHJLBTNAGRQEKS-UHFFFAOYSA-M 0.000 description 6
- 238000003860 storage Methods 0.000 description 6
- QGJOPFRUJISHPQ-UHFFFAOYSA-N Carbon disulfide Chemical compound S=C=S QGJOPFRUJISHPQ-UHFFFAOYSA-N 0.000 description 5
- 102000003952 Caspase 3 Human genes 0.000 description 5
- 108090000397 Caspase 3 Proteins 0.000 description 5
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 5
- 229910004042 HAuCl4 Inorganic materials 0.000 description 5
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 5
- 229910018885 Pt—Au Inorganic materials 0.000 description 5
- 241000283984 Rodentia Species 0.000 description 5
- 238000003917 TEM image Methods 0.000 description 5
- 238000004833 X-ray photoelectron spectroscopy Methods 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- 150000007513 acids Chemical class 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 239000011258 core-shell material Substances 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- 238000003487 electrochemical reaction Methods 0.000 description 5
- 238000011065 in-situ storage Methods 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 210000004072 lung Anatomy 0.000 description 5
- 238000012544 monitoring process Methods 0.000 description 5
- 229910017604 nitric acid Inorganic materials 0.000 description 5
- 230000003204 osmotic effect Effects 0.000 description 5
- KIDPOJWGQRZHFM-UHFFFAOYSA-N platinum;hydrate Chemical compound O.[Pt] KIDPOJWGQRZHFM-UHFFFAOYSA-N 0.000 description 5
- 229910000027 potassium carbonate Inorganic materials 0.000 description 5
- 238000005086 pumping Methods 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 229910000029 sodium carbonate Inorganic materials 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 230000003595 spectral effect Effects 0.000 description 5
- 238000001228 spectrum Methods 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 208000012766 Growth delay Diseases 0.000 description 4
- 229920002594 Polyethylene Glycol 8000 Polymers 0.000 description 4
- 101100004606 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) BPH1 gene Proteins 0.000 description 4
- 150000001450 anions Chemical class 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 230000002210 biocatalytic effect Effects 0.000 description 4
- 230000005540 biological transmission Effects 0.000 description 4
- 150000001768 cations Chemical class 0.000 description 4
- 230000004663 cell proliferation Effects 0.000 description 4
- 210000003169 central nervous system Anatomy 0.000 description 4
- 210000003679 cervix uteri Anatomy 0.000 description 4
- 238000004140 cleaning Methods 0.000 description 4
- 239000012141 concentrate Substances 0.000 description 4
- 238000011109 contamination Methods 0.000 description 4
- 238000010924 continuous production Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000010894 electron beam technology Methods 0.000 description 4
- 238000001704 evaporation Methods 0.000 description 4
- 230000008020 evaporation Effects 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 239000012467 final product Substances 0.000 description 4
- 230000005484 gravity Effects 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 238000000608 laser ablation Methods 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- 230000011278 mitosis Effects 0.000 description 4
- 230000036961 partial effect Effects 0.000 description 4
- 229920003229 poly(methyl methacrylate) Polymers 0.000 description 4
- 229920000515 polycarbonate Polymers 0.000 description 4
- 239000008213 purified water Substances 0.000 description 4
- 238000003608 radiolysis reaction Methods 0.000 description 4
- 230000019491 signal transduction Effects 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 230000001052 transient effect Effects 0.000 description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 4
- 235000019801 trisodium phosphate Nutrition 0.000 description 4
- 238000012795 verification Methods 0.000 description 4
- 210000003905 vulva Anatomy 0.000 description 4
- 239000002699 waste material Substances 0.000 description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 3
- 239000007836 KH2PO4 Substances 0.000 description 3
- 239000002841 Lewis acid Substances 0.000 description 3
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 3
- VVQNEPGJFQJSBK-UHFFFAOYSA-N Methyl methacrylate Chemical compound COC(=O)C(C)=C VVQNEPGJFQJSBK-UHFFFAOYSA-N 0.000 description 3
- 101000930477 Mus musculus Albumin Proteins 0.000 description 3
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 3
- 229920005372 Plexiglas® Polymers 0.000 description 3
- 239000007983 Tris buffer Substances 0.000 description 3
- 238000001994 activation Methods 0.000 description 3
- 230000001640 apoptogenic effect Effects 0.000 description 3
- XKRFYHLGVUSROY-UHFFFAOYSA-N argon Substances [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 150000007516 brønsted-lowry acids Chemical class 0.000 description 3
- 239000006227 byproduct Substances 0.000 description 3
- 239000001110 calcium chloride Substances 0.000 description 3
- 229910001628 calcium chloride Inorganic materials 0.000 description 3
- 239000003990 capacitor Substances 0.000 description 3
- 230000022131 cell cycle Effects 0.000 description 3
- 239000013626 chemical specie Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 229940125904 compound 1 Drugs 0.000 description 3
- 229940125782 compound 2 Drugs 0.000 description 3
- 230000003750 conditioning effect Effects 0.000 description 3
- 239000010949 copper Substances 0.000 description 3
- 238000002425 crystallisation Methods 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 238000007405 data analysis Methods 0.000 description 3
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 3
- 229910000397 disodium phosphate Inorganic materials 0.000 description 3
- 235000019800 disodium phosphate Nutrition 0.000 description 3
- 239000002270 dispersing agent Substances 0.000 description 3
- 238000012377 drug delivery Methods 0.000 description 3
- 230000009977 dual effect Effects 0.000 description 3
- 239000012777 electrically insulating material Substances 0.000 description 3
- 230000005672 electromagnetic field Effects 0.000 description 3
- 230000009881 electrostatic interaction Effects 0.000 description 3
- 239000012149 elution buffer Substances 0.000 description 3
- 230000004907 flux Effects 0.000 description 3
- 230000005251 gamma ray Effects 0.000 description 3
- 229920001903 high density polyethylene Polymers 0.000 description 3
- 239000004700 high-density polyethylene Substances 0.000 description 3
- 230000002706 hydrostatic effect Effects 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 229910010272 inorganic material Inorganic materials 0.000 description 3
- 239000011147 inorganic material Substances 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 150000007517 lewis acids Chemical class 0.000 description 3
- 238000013507 mapping Methods 0.000 description 3
- 239000010445 mica Substances 0.000 description 3
- 229910052618 mica group Inorganic materials 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 3
- 235000019799 monosodium phosphate Nutrition 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 210000001672 ovary Anatomy 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 230000002572 peristaltic effect Effects 0.000 description 3
- 230000000704 physical effect Effects 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 235000015497 potassium bicarbonate Nutrition 0.000 description 3
- 235000011181 potassium carbonates Nutrition 0.000 description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 3
- 235000011009 potassium phosphates Nutrition 0.000 description 3
- 230000000717 retained effect Effects 0.000 description 3
- 230000002441 reversible effect Effects 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 235000017550 sodium carbonate Nutrition 0.000 description 3
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 3
- 210000000952 spleen Anatomy 0.000 description 3
- 210000003932 urinary bladder Anatomy 0.000 description 3
- 210000004291 uterus Anatomy 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 230000005653 Brownian motion process Effects 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 229910002621 H2PtCl6 Inorganic materials 0.000 description 2
- 229910003594 H2PtCl6.6H2O Inorganic materials 0.000 description 2
- 108091006905 Human Serum Albumin Proteins 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 239000004743 Polypropylene Substances 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 2
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 2
- 229920006397 acrylic thermoplastic Polymers 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- HSFWRNGVRCDJHI-UHFFFAOYSA-N alpha-acetylene Natural products C#C HSFWRNGVRCDJHI-UHFFFAOYSA-N 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- 230000002238 attenuated effect Effects 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 235000012206 bottled water Nutrition 0.000 description 2
- 238000005537 brownian motion Methods 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000002800 charge carrier Substances 0.000 description 2
- 229910052729 chemical element Inorganic materials 0.000 description 2
- XTEGARKTQYYJKE-UHFFFAOYSA-N chloric acid Chemical compound OCl(=O)=O XTEGARKTQYYJKE-UHFFFAOYSA-N 0.000 description 2
- 229940005991 chloric acid Drugs 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 230000008025 crystallization Effects 0.000 description 2
- 238000013480 data collection Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000002242 deionisation method Methods 0.000 description 2
- 229910052805 deuterium Inorganic materials 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000012538 diafiltration buffer Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000010411 electrocatalyst Substances 0.000 description 2
- 230000005518 electrochemistry Effects 0.000 description 2
- 230000002124 endocrine Effects 0.000 description 2
- 230000003628 erosive effect Effects 0.000 description 2
- 125000002534 ethynyl group Chemical group [H]C#C* 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000008570 general process Effects 0.000 description 2
- 239000007952 growth promoter Substances 0.000 description 2
- 239000001307 helium Substances 0.000 description 2
- 229910052734 helium Inorganic materials 0.000 description 2
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 2
- 150000002431 hydrogen Chemical class 0.000 description 2
- 229910052806 inorganic carbonate Inorganic materials 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 150000001455 metallic ions Chemical class 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000006199 nebulizer Substances 0.000 description 2
- 229910052754 neon Inorganic materials 0.000 description 2
- GKAOGPIIYCISHV-UHFFFAOYSA-N neon atom Chemical compound [Ne] GKAOGPIIYCISHV-UHFFFAOYSA-N 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 238000001139 pH measurement Methods 0.000 description 2
- 230000036314 physical performance Effects 0.000 description 2
- 150000003057 platinum Chemical class 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 239000004926 polymethyl methacrylate Substances 0.000 description 2
- 229920001155 polypropylene Polymers 0.000 description 2
- 229920002223 polystyrene Polymers 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 230000006920 protein precipitation Effects 0.000 description 2
- 239000010453 quartz Substances 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000000310 rehydration solution Substances 0.000 description 2
- 239000012266 salt solution Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000000851 scanning transmission electron micrograph Methods 0.000 description 2
- 229910052710 silicon Inorganic materials 0.000 description 2
- 239000010703 silicon Substances 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- SDKPSXWGRWWLKR-UHFFFAOYSA-M sodium;9,10-dioxoanthracene-1-sulfonate Chemical compound [Na+].O=C1C2=CC=CC=C2C(=O)C2=C1C=CC=C2S(=O)(=O)[O-] SDKPSXWGRWWLKR-UHFFFAOYSA-M 0.000 description 2
- 238000004544 sputter deposition Methods 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 2
- ISXSCDLOGDJUNJ-UHFFFAOYSA-N tert-butyl prop-2-enoate Chemical compound CC(C)(C)OC(=O)C=C ISXSCDLOGDJUNJ-UHFFFAOYSA-N 0.000 description 2
- 229940071240 tetrachloroaurate Drugs 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 239000010409 thin film Substances 0.000 description 2
- 210000001685 thyroid gland Anatomy 0.000 description 2
- 230000036962 time dependent Effects 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 230000003442 weekly effect Effects 0.000 description 2
- TUSDEZXZIZRFGC-UHFFFAOYSA-N 1-O-galloyl-3,6-(R)-HHDP-beta-D-glucose Natural products OC1C(O2)COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC1C(O)C2OC(=O)C1=CC(O)=C(O)C(O)=C1 TUSDEZXZIZRFGC-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- ZOOGRGPOEVQQDX-UUOKFMHZSA-N 3',5'-cyclic GMP Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=C(NC2=O)N)=C2N=C1 ZOOGRGPOEVQQDX-UUOKFMHZSA-N 0.000 description 1
- HJCMDXDYPOUFDY-WHFBIAKZSA-N Ala-Gln Chemical compound C[C@H](N)C(=O)N[C@H](C(O)=O)CCC(N)=O HJCMDXDYPOUFDY-WHFBIAKZSA-N 0.000 description 1
- 229910000952 Be alloy Inorganic materials 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- 241000282994 Cervidae Species 0.000 description 1
- 241000283153 Cetacea Species 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- 238000012287 DNA Binding Assay Methods 0.000 description 1
- 239000001263 FEMA 3042 Substances 0.000 description 1
- YCKRFDGAMUMZLT-UHFFFAOYSA-N Fluorine atom Chemical compound [F] YCKRFDGAMUMZLT-UHFFFAOYSA-N 0.000 description 1
- 230000010190 G1 phase Effects 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 238000012404 In vitro experiment Methods 0.000 description 1
- 229910020252 KAuCl4 Inorganic materials 0.000 description 1
- 241000219739 Lens Species 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 230000010757 Reduction Activity Effects 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 230000018199 S phase Effects 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 206010041067 Small cell lung cancer Diseases 0.000 description 1
- 229920002125 Sokalan® Polymers 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 229920006362 Teflon® Polymers 0.000 description 1
- ZMZDMBWJUHKJPS-UHFFFAOYSA-M Thiocyanate anion Chemical compound [S-]C#N ZMZDMBWJUHKJPS-UHFFFAOYSA-M 0.000 description 1
- 208000024770 Thyroid neoplasm Diseases 0.000 description 1
- 102000004338 Transferrin Human genes 0.000 description 1
- 108090000901 Transferrin Proteins 0.000 description 1
- 208000002495 Uterine Neoplasms Diseases 0.000 description 1
- 208000004354 Vulvar Neoplasms Diseases 0.000 description 1
- 238000000026 X-ray photoelectron spectrum Methods 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 239000003570 air Substances 0.000 description 1
- 229960002648 alanylglutamine Drugs 0.000 description 1
- 238000011166 aliquoting Methods 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 239000003945 anionic surfactant Substances 0.000 description 1
- 230000005735 apoptotic response Effects 0.000 description 1
- 229940072107 ascorbate Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 235000021053 average weight gain Nutrition 0.000 description 1
- 238000003705 background correction Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 229910001423 beryllium ion Inorganic materials 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 239000012490 blank solution Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 230000004700 cellular uptake Effects 0.000 description 1
- PBAYDYUZOSNJGU-UHFFFAOYSA-N chelidonic acid Natural products OC(=O)C1=CC(=O)C=C(C(O)=O)O1 PBAYDYUZOSNJGU-UHFFFAOYSA-N 0.000 description 1
- HGAZMNJKRQFZKS-UHFFFAOYSA-N chloroethene;ethenyl acetate Chemical compound ClC=C.CC(=O)OC=C HGAZMNJKRQFZKS-UHFFFAOYSA-N 0.000 description 1
- 229940105442 cisplatin injection Drugs 0.000 description 1
- 239000010415 colloidal nanoparticle Substances 0.000 description 1
- 238000002485 combustion reaction Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 239000012468 concentrated sample Substances 0.000 description 1
- 239000004020 conductor Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 230000000368 destabilizing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000011038 discontinuous diafiltration by volume reduction Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000002612 dispersion medium Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 238000010292 electrical insulation Methods 0.000 description 1
- 238000009429 electrical wiring Methods 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 239000007772 electrode material Substances 0.000 description 1
- 230000005670 electromagnetic radiation Effects 0.000 description 1
- 238000000609 electron-beam lithography Methods 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 238000002149 energy-dispersive X-ray emission spectroscopy Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000005530 etching Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000002657 fibrous material Substances 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000000705 flame atomic absorption spectrometry Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 238000001640 fractional crystallisation Methods 0.000 description 1
- 239000003574 free electron Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 208000005017 glioblastoma Diseases 0.000 description 1
- 150000002343 gold Chemical class 0.000 description 1
- 229940111120 gold preparations Drugs 0.000 description 1
- RJHLTVSLYWWTEF-UHFFFAOYSA-K gold trichloride Chemical class Cl[Au](Cl)Cl RJHLTVSLYWWTEF-UHFFFAOYSA-K 0.000 description 1
- CBMIPXHVOVTTTL-UHFFFAOYSA-N gold(3+) Chemical compound [Au+3] CBMIPXHVOVTTTL-UHFFFAOYSA-N 0.000 description 1
- 230000036449 good health Effects 0.000 description 1
- 230000005283 ground state Effects 0.000 description 1
- 239000003673 groundwater Substances 0.000 description 1
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 238000000000 high-resolution scanning transmission electron microscopy Methods 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 1
- ZMZDMBWJUHKJPS-UHFFFAOYSA-N hydrogen thiocyanate Natural products SC#N ZMZDMBWJUHKJPS-UHFFFAOYSA-N 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000009616 inductively coupled plasma Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 230000002601 intratumoral effect Effects 0.000 description 1
- 238000010884 ion-beam technique Methods 0.000 description 1
- 231100001231 less toxic Toxicity 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 238000003754 machining Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000009285 membrane fouling Methods 0.000 description 1
- 150000001247 metal acetylides Chemical class 0.000 description 1
- 229910001092 metal group alloy Inorganic materials 0.000 description 1
- 229910052987 metal hydride Inorganic materials 0.000 description 1
- 150000004681 metal hydrides Chemical class 0.000 description 1
- 239000007769 metal material Substances 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- 239000013528 metallic particle Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000008450 motivation Effects 0.000 description 1
- IXEGRINNWXKNJO-UHFFFAOYSA-N n-hexadecylaniline Chemical compound CCCCCCCCCCCCCCCCNC1=CC=CC=C1 IXEGRINNWXKNJO-UHFFFAOYSA-N 0.000 description 1
- 239000011234 nano-particulate material Substances 0.000 description 1
- 210000003757 neuroblast Anatomy 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 150000004767 nitrides Chemical class 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000002736 nonionic surfactant Substances 0.000 description 1
- 229910052755 nonmetal Inorganic materials 0.000 description 1
- QYSGYZVSCZSLHT-UHFFFAOYSA-N octafluoropropane Chemical compound FC(F)(F)C(F)(F)C(F)(F)F QYSGYZVSCZSLHT-UHFFFAOYSA-N 0.000 description 1
- 238000005580 one pot reaction Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 210000004303 peritoneum Anatomy 0.000 description 1
- 238000006552 photochemical reaction Methods 0.000 description 1
- 238000000206 photolithography Methods 0.000 description 1
- 238000001020 plasma etching Methods 0.000 description 1
- 238000009832 plasma treatment Methods 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000004584 polyacrylic acid Substances 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 239000004814 polyurethane Substances 0.000 description 1
- 229920002635 polyurethane Polymers 0.000 description 1
- 239000004800 polyvinyl chloride Substances 0.000 description 1
- 230000003334 potential effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000026938 proteasomal ubiquitin-dependent protein catabolic process Effects 0.000 description 1
- 239000011253 protective coating Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000012846 protein folding Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 239000011819 refractory material Substances 0.000 description 1
- 230000001846 repelling effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000005060 rubber Substances 0.000 description 1
- 238000010079 rubber tapping Methods 0.000 description 1
- 229910052701 rubidium Inorganic materials 0.000 description 1
- 238000005464 sample preparation method Methods 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- LRBQNJMCXXYXIU-NRMVVENXSA-N tannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-NRMVVENXSA-N 0.000 description 1
- 229940033123 tannic acid Drugs 0.000 description 1
- 235000015523 tannic acid Nutrition 0.000 description 1
- 229920002258 tannic acid Polymers 0.000 description 1
- 231100001274 therapeutic index Toxicity 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000012876 topography Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000012581 transferrin Substances 0.000 description 1
- 229910001428 transition metal ion Inorganic materials 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000009834 vaporization Methods 0.000 description 1
- 230000008016 vaporization Effects 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 238000004804 winding Methods 0.000 description 1
- 229910052724 xenon Inorganic materials 0.000 description 1
- FHNFHKCVQCLJFQ-UHFFFAOYSA-N xenon atom Chemical compound [Xe] FHNFHKCVQCLJFQ-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/243—Platinum; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/242—Gold; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/143—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1611—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82B—NANOSTRUCTURES FORMED BY MANIPULATION OF INDIVIDUAL ATOMS, MOLECULES, OR LIMITED COLLECTIONS OF ATOMS OR MOLECULES AS DISCRETE UNITS; MANUFACTURE OR TREATMENT THEREOF
- B82B1/00—Nanostructures formed by manipulation of individual atoms or molecules, or limited collections of atoms or molecules as discrete units
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82B—NANOSTRUCTURES FORMED BY MANIPULATION OF INDIVIDUAL ATOMS, MOLECULES, OR LIMITED COLLECTIONS OF ATOMS OR MOLECULES AS DISCRETE UNITS; MANUFACTURE OR TREATMENT THEREOF
- B82B3/00—Manufacture or treatment of nanostructures by manipulation of individual atoms or molecules, or limited collections of atoms or molecules as discrete units
- B82B3/0009—Forming specific nanostructures
-
- C—CHEMISTRY; METALLURGY
- C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
- C22C—ALLOYS
- C22C1/00—Making non-ferrous alloys
- C22C1/04—Making non-ferrous alloys by powder metallurgy
- C22C1/0466—Alloys based on noble metals
-
- C—CHEMISTRY; METALLURGY
- C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
- C22C—ALLOYS
- C22C30/00—Alloys containing less than 50% by weight of each constituent
-
- C—CHEMISTRY; METALLURGY
- C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
- C22C—ALLOYS
- C22C5/00—Alloys based on noble metals
- C22C5/02—Alloys based on gold
-
- C—CHEMISTRY; METALLURGY
- C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
- C22C—ALLOYS
- C22C5/00—Alloys based on noble metals
- C22C5/04—Alloys based on a platinum group metal
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Definitions
- the present invention relates to novel gold-platinum based bi-metallic nanocrystal suspensions that have nanocrystal surfaces that are substantially free from organic or other impurities or films associated with typical chemical reductants/stabilizers and/or raw materials used in nanoparticle formation processes.
- the surfaces are “clean” relative to the surfaces of metal-based nanoparticles made using chemical reduction (and other) processes that require organic (or other) reductants and/or surfactants to grow (and/or suspend) metal nanoparticles from metal ions in a solution.
- the invention includes novel electrochemical manufacturing apparatuses and techniques for making the bi-metallic nanocrystal suspensions.
- the techniques do not require the use or presence of chlorine ions/atoms and/or chlorides or chlorine-based materials for the manufacturing process/final suspension.
- the invention further includes pharmaceutical compositions thereof and the use of the bi-metallic nanocrystals or suspensions or colloids thereof for the treatment or prevention of diseases or conditions for which metal-based therapy is already known, including, for example, for cancerous diseases or conditions.
- nanometals exhibit a variety of electronic, optical, magnetic and/or chemical properties which are typically not achievable when metallic materials are in their bulk form.
- metals that are relatively inert at the macroscale such as platinum and gold, are excellent catalysts at the nanoscale.
- combinations of two different metals (bi-metallic) at the nanoscale offer further interesting performance issues.
- the different metals may result in mixtures of metals, alloys or heterogeneous structures, each of which my exhibit different physical properties and/or performance characteristics.
- Applications for bi-metallic nanoparticulate metals include electronics and computing devices, bionanotechnology, medical treatment and diagnosis and energy generation and storage. The use of these bi-metallic nanometals for a variety of applications requires efficient and safe approaches for manufacturing such materials.
- top-down and bottom-up approaches two fundamentally different approaches have been used to manufacture bi-metallic nanomaterials and they are referred to as “top-down” and “bottom-up” approaches.
- top-down approach bi-metallic nanomaterials are manufactured from larger entities typically, without atomic-level control.
- Typcial top-down approaches include such techniques as photolithography and electron-beam lithography which start with large materials and use either machining or etching techniques to make small materials.
- Laser ablation is also a known top-down approach.
- bi-metallic nanomaterials are manufactured from two or more molecular components which are caused to be assembled into bi-metallic nanoparticulate materials.
- building blocks are first formed and then the building blocks are assembled into a final nano-material.
- bi-metallic approaches include templating, chemical synthesis, sonochemical approaches, electrochemical approaches, sonoelectrochemical approaches, thermal and photochemical reduction methods including ⁇ -ray, x-ray, laser and microwave, each of which has certain negative process and/or product limitations associated therewith.
- bi-metallic nanoparticles In the case where two metals are formed into bi-metallic nanoparticles, further considerations such as whether the bi-metallic nanoparticles are alloys, partial alloys or partially phase segregated or completely phase segregated are also important because the specific configuration of the nanoparticles can result in different performance (e.g., biologic or catalytic).
- Faraday used reduction chemistry techniques to reduce chemically an aqueous gold salt, chloroaurate (i.e., a gold (III) salt), utilizing either phosphorous dispersed into ether (e.g., CH 3 —CH 2 —O—CH 2 —CH 3 ), or carbon disulfide (i.e, CS 2 ), as the reductant.
- chloroaurate i.e., a gold (III) salt
- phosphorous dispersed into ether e.g., CH 3 —CH 2 —O—CH 2 —CH 3
- carbon disulfide i.e, CS 2
- gold (III) complex such as hydrogen tetrachloroaurate (or chloric acid) and reduce the gold in the gold complex to gold metal (i.e., gold (0) or metallic gold) by using added chemical species reductants, such as Na thiocyanate, White P, Na 3 citrate & tannic acid, NaBH 4 , Citric Acid, Ethanol, Na ascorbate, Na 3 citrate, Hexadecylaniline and others (Brown, 2008).
- chemical species reductants such as Na thiocyanate, White P, Na 3 citrate & tannic acid, NaBH 4 , Citric Acid, Ethanol, Na ascorbate, Na 3 citrate, Hexadecylaniline and others (Brown, 2008).
- Bi-metallic nanocrystals have been formed by a number of different techniques including forming nanoparticles from the solid, gaseous and solution states.
- the solid state typically requires high temperature heating and annealing.
- the typical gaseous state approaches usually utilize molecular beam techniques, namely, the vaporization of mixed metallic powder by lasers, pulsed-arc beams, etc.
- the solution state is the much more heavily utilized bi-metallic nanoparticle formation technique.
- the proper chemical reactants e.g., metal-based salts and reductants and/or stabilizers
- proper control of certain intermediate reactions which can or do occur
- control of corresponding crystallization reactions are required to achieve desired metallic nanoparticles (Wang, 2011).
- bi-metallic nanocrystals can be achieved such as a core/shell (also known as a hetero-aggregate), a hetero-structure or hetero-aggregate, an intermetallic, a mixture or alloy, as well as various core shell arrangements (Wanjala, 2011). All of these different types of bi-metallic nanocrystals can have quite different physical performance capabilities.
- nanoparticle surface chemistry Since the importance of nanoparticle surface chemistry is now beginning to be focused on as a key for understanding and controlling nanoparticle performance issues, attempts are now being made to remove constituents associated with manufacturing processes that are located on the surface of the formed nanoparticle (e.g., the outer layer or the presence of constituents formed as a result of reducing agent and/or surface capping agent and/or other raw materials used) including going so far as utilizing an oxygen plasma combined with electrochemical stripping (Yang, 2011). However, such surface modification approaches result in their own changes to the nanoparticle surface.
- the reductant surface coating or film is permitted to remain as an impurity on the surface of the nanoparticles, but in other cases, it is attempted to be removed by a variety of somewhat complex and costly techniques. When removed, the coating typically is replaced by an alternative composition or coating to permit the nanoparticles to stay in suspension when hydrated.
- the influence of surface purity on the chemistry and properties of nanoparticles is often overlooked; however, results now indicate that the extent of purification can have a significant impact (Sweeney, 2006).
- These researchers noted that sufficient purification of nanoparticles can be more challenging that the preparation itself, usually involving tedious, time-consuming and wasteful procedures such as extensive solvent washes and fractional crystallization. Absent such purification, the variables of surface chemistry-related contaminants on the surface of chemically reduced nanoparticles affects the ability to understand/control basic structure-function relationships (Sweeney, 2006).
- Subsequent processing techniques may also require a set of washing steps, certain concentrating or centrifuging steps, and/or subsequent chemical reaction coating steps, all of which are required to achieve desirable results and certain performance characteristics (e.g., stabilization due to ligand exchange, efficacy, etc.) for the nanoparticles and nanoparticle suspensions (Sperling, 2008).
- certain performance characteristics e.g., stabilization due to ligand exchange, efficacy, etc.
- harsh stripping methods are used to ensure very clean nanoparticle surfaces (Panyala, 2009).
- a surface coating comprising one or more elements of the reductant and/or the surfactant or capping agent will be present on (or in) at least a portion of the suspended nanoparticles.
- a reductant i.e., a reducing agent
- the reducing agent coating or surface impurity is sometimes added to or even replaced by surfactant coatings or capping agents.
- reductant/surfactant coatings or films can be viewed as impurities located on and/or in the metal-based nanoparticles and may result in such colloids or sols actually possessing more of the properties of the protective coating or film than the nanoparticle per se (Weiser, p. 42, 1933).
- surfactants and amphiphilic polymers become heavily involved not only in the formation of nanoparticles (thus affecting size and shape), but also in the nanoparticles per se.
- Surface properties of the nanoparticles are modified by reductant coatings and/or surfactant molecule coatings (Sperling, 2008).
- Sonoelectrical processes typically direct electric and acoustic energy toward metal-based raw material salts (e.g., HAuCl 4 ⁇ 4H 2 O (AuCl 4 ⁇ ), NaAuCl 4 ⁇ 2H 2 O, H 2 PtCl 6 ⁇ 6H 2 O, HAuCl 3 ⁇ 3H 2 O, etc.) and metal ions in those salts are caused to be reduced by one or more reductant species created by the sonoelectrochemial method.
- metal-based raw material salts e.g., HAuCl 4 ⁇ 4H 2 O (AuCl 4 ⁇ ), NaAuCl 4 ⁇ 2H 2 O, H 2 PtCl 6 ⁇ 6H 2 O, HAuCl 3 ⁇ 3H 2 O, etc.
- a single electrode induces the growth of nanoparticles thereon by an electrochemical step, followed by an acoustic step which, more or less, attempts to eject the nanoparticles off from the electrode and also creates additional reductant material by, for example, lysis of water molecules.
- a single electrode typically performs a dual duty of both electrochemistry (e.g., nanoparticle formation) and acoustic chemistry (e.g., reductant formation) (Nagata, 1996).
- sonoelectrochemical techniques utilize one or more reductants and/or capping agents in addition to any of those which may be formed in situ by the process.
- a variety of different polymers have been utilized as capping agents for single metallic nanoparticles (Saez, 2009).
- work by others (Liu, 2004; Ou, 2011; Mai, 2011; and Liu, 2006) all disclose similar sonoelectrochemical techniques for making gold nanoparticles with sonoelectrochemical pulse methods using, allegedly, no added reductants.
- utilization of an acid solution in combination with electrochemical cycling to strip gold ions from a gold electrode and form AuCl 4 ⁇ compounds in an aqueous solution has been disclosed (Liu, 2004).
- the gold ions are reduced by created reductant species (e.g., lysis products of H 2 O) produced in their sonoelectrochemical process.
- reductant species e.g., lysis products of H 2 O
- concentrations of gold nanoparticles produced are quite limited by this technique (e.g., 3 ppm) without the addition of other materials (e.g., stabilizers) (Ou, 2011).
- a variety of sonoelectrochemical technquies have also been set forth for making bi-metallic nanoparticles.
- platinum-gold nanoparticles stabilized by PEG-MS polyetholeneglycolmonostearate
- binary gold/platinum nanoparticles made by sonoelectrochemistry utilizing surfactants anionic surfactants; sodium dodechal sulfate (SDS) or nonionic surfactant polyetholeneglycolmonostearate PEG-MS
- surfactants anionic surfactants; sodium dodechal sulfate (SDS) or nonionic surfactant polyetholeneglycolmonostearate PEG-MS
- Radiolytic techniques for making nanoparticles have been directed primarily to single-metals (i.e., not bi-metals).
- Another older and more complex technique for minimizing or eliminating the need for reducing agents and/or minimizing undesirable oxidation products of the reductant utilizes ⁇ -irradiation from a 60 Co source at a dose rate of 1.8 ⁇ 10 4 rad/h.
- Au(CN) 2 was reduced by first creating hydrated electrons from the radiolysis of water and utilizing the hydrated electrons to reduce the gold ions, namely:
- Futher the creation of hydrated electrons and OH radicals by pulse activation from a linear accelerator has also occurred (Ghosh-Mazumdar, 1968). Such created species assist in the reduction of various metals from aqueous metallic-based salts.
- a one-pot synthesis of Au—Pt alloys by intense x-ray irradiation has also been disclosed (Wang, 2011).
- the incident x-rays irradiate a gold/platinum salt solution (i.e., HAuCl 4 ⁇ 3H 2 O and H 2 PtCl 6 .6H 2 O) containing PEG (a common surfactant molecule known to prevent nanoparticle aggregation).
- PEG a common surfactant molecule known to prevent nanoparticle aggregation.
- PEG could negatively impact applications that are sensitive to surface conditions, such as catalysis (Wang, 2011).
- Bi-metallic Pt—Au nanoparticles have been made by femtosecond laser synthesis (Chau, 2011). Specifically, gold and platinum salt solutions (i.e., HAuCl 4 ⁇ 4H 2 O, H 2 PtCl 6 .6H 2 O) were combined with PVP (a known dispersing/stabilizing agent) and the solution was laser irradiated. In related work, high intensity laser radiation of a similar solution of gold and platinum salts occurred. However, in this solution no PEG was added and the resultant nanoparticles were found not to be stable (Nakamura, 2011; Nakamura, 2010; Nakamura, 2009).
- PVP a known dispersing/stabilizing agent
- Bi-metallic gold-platinum nanoparticles have also been made by electron beam irradiation (Mirdamadi-Esfahani, 2010). Specifically, in this approach, the electron beam irradiation creates hydrated electrons and reducing radicals due to the radiolysis of water. Metal salts of gold and platinum (i.e., KAuCl 4 and H 2 PtCl 6 ) are mixed with polyacrylic acid (i.e., a dispersant/stabilizing agent) and accelerated electrons are directed thereto.
- KAuCl 4 and H 2 PtCl 6 Metal salts of gold and platinum
- polyacrylic acid i.e., a dispersant/stabilizing agent
- Different surface chemistries or surface films can result in different interactions of the nanoparticles with, for example, a variety of proteins in an organism.
- Biophysical binding forces e.g., electrostatic, hydrophobic, hydrogen binding, van der Waals
- of nanoparticles to proteins are a function not only of the size, shape and composition of the nanoparticles, but also the type of and/or thickness of the surface impurities or coating(s) on the nanoparticles (Lacerda, 2010).
- Nanoparticles A better understanding of the biological effects of nanoparticles requires an understanding of the binding properties of the in-vivo proteins that associate themselves with the nanoparticles. Protein absorption (or a protein corona) on nanoparticles can change as a function of nanoparticle size and surface layer composition and thickness. Protein layers that “dress” the nanoparticle control the propensity of the nanoparticles to aggregate and strongly influence their interaction with biological materials (Lacerda, 2010).
- CTAB-coated gold nanoparticles release portions of their coatings at different points in a biological process and/or different location(s) within an organism, which results in toxicity (Qui, 2010).
- the protein corona which forms on a nanoparticle is important because it is the protein corona that gives the biological identity to the nanoparticle (Lynch, 2007).
- the surface of the nanoparticle assists in the formation of the protein corona as well as its size and its shape (Lynch, 2007).
- albumin-based drug delivery has been recognized as a novel therapeutic approach (Wunder, 2003; Stehle, 1997; Stehle, 1997). Specifically, the albumin-binding assists in delivery of the therapeutic to desirable targeted locations resulting in higher efficacy/lower toxicity.
- New bi-metallic nanocrystal suspensions are provided that have nanocrystalline surfaces that can be substantially free (as defined herein) from organic or other impurities or films, or in certain cases may contain some desirable film or partial coating. Specifically, the surfaces are “clean” relative to those made using chemical reduction processes that require chemical reductants and/or surfactants to grow gold nanoparticles from metal ions in solution.
- Resulting bi-metallic nanocrystalline suspensions or colloids have desirable pH ranges such as 4.0-12.0, but more typically 5.0-11.0, and even more typically 8.0-11.0, and in many embodiment, 10.0-11.0 and zeta potential values of at least ⁇ 20 mV, and more typically at least ⁇ 40 mV, and even more typically at least ⁇ 50 mV for the pH ranges of interest.
- shapes and shape distributions of these bi-metallic nanocrystals prepared according to the manufacturing process described below include, but are not limited to, spheres, pentagons, hexagons (e.g., hexagonal bipyramids, icosahedrons, octahedrons), and “others”.
- any desired average size of bi-metallic nanocrystals below 100 nm can be provided.
- the most desirable crystalline size ranges include those having an average crystal size (as measured and determined by specific techniques disclosed in detail herein) that is predominantly less than 100 nm, and more typically less than 50 nm, even more typically less than 30 nm, and in many of the preferred embodiments disclosed herein, the average crystal size for the nanocrystal size distribution is less than 20 nm and with an even more preferable range of 8-18 nm.
- the electrochemical techniques disclosed herein can be utilized to result in larger nanocrystals, if desired.
- concentrations of bi-metallic nanocrystals can be provided according to the invention.
- total atomic metal concentrations of bi-metallic nanocrystals produced initially can be a few parts per million (i.e., ⁇ g/ml or mg/l) up to a few hundred ppm, but are typically in the range of 2-200 ppm (i.e., 2 ⁇ g/ml-200 ⁇ g/ml) and more often in the range of 2-50 ppm (i.e., 2 ⁇ g/ml-50 ⁇ g/ml) and even more typically 5-20 ppm (i.e., 5 ⁇ g/ml-20 ⁇ g/ml).
- novel concentration techniques are disclosed herein which allow concentrated “initial” product to be formed with ppm's between 200-5,000 ppm and more preferably, 200-3,000 ppm and more preferably, 200-1,000 ppm.
- the bi-metallic nanocrystals in suspension can be made as alloys, partial alloys, phase-segregated or heteroaggregates or mixtures.
- the bi-metallic nanocrystals are alloys and/or heteroaggregates.
- Gold is typically the major constituent (i.e., more by weight and more by volume) and platinum is typically the minor constituent (i.e., less by weight and less by volume).
- Typical ratios range from 2/1 to 10/1, with preferred ranges being 3/1 to 8/1, and even more preferred 3/1 to 6/1.
- a novel set of processes are provided to produce these unique bi-metallic nanocrystals.
- Each process involves the creation of the bi-metallic nanocrystals in water.
- the water contains an added “process enhancer” which does not significantly bind to the formed nanocrystals, but rather facilitates nucleation/crystal growth during the electrochemical-stimulated growth process.
- the process enhancer serves important roles in the process including, for example, providing charged ions in the electrochemical solution to permit the crystals to be grown.
- a first step includes forming a platinum metal-based species with at least one process enhancer and the formed aqueous suspension/solution is then used as a raw material solution/suspension in a second step where a gold metal-based species is reduced and/or co-reduced to grow the bi-metallic nanocrystals in water.
- the processes involve first forming electrochemically at least one platinum species in water and at least one lysis product of water, thereby creating a platinum species and water material; and using the created platinum/water material in a second electrochemical reaction to form a suspension of bi-metallic gold-platinum nanocrystals in water.
- these bi-metallic nanocrystals can form alloys or metal “coatings” (or portions of coatings, e.g., islands) on core metals or alternatively, form heteroaggregates.
- a mixture of nanocrystals can be made.
- a range of alloys or mixtures or heteroaggregates may result within a single colloid or suspension, if desired.
- desirable residual metal ions may be in solution in the suspension.
- novel electrochemical processes can occur in either a batch, semi-continuous or continuous process. These processes result in controlled bi-metallic nanocrystalline concentrations, controlled nanocrystal sizes and controlled nanocrystal size ranges. Novel manufacturing assemblies are provided to produce these bi-metallic nanocrystals.
- bi-metallic nanocrystals have substantially cleaner surfaces than the prior available metallic-based (or bi-metallic-based) nanoparticles, and can desirably contain spatially extended low index crystallographic planes forming novel crystal shapes and/or crystal shape distributions
- the bi-metallic nanocrystals appear to be more active (e.g., more biologically active and may be less toxic) relative to those containing surface contaminants such as chemical reductants and/or surfactants or residual raw materials that result from traditional chemical reduction (or other) processes. Therefore, uses for nanoparticles, such as, catalysis processes, medical treatments, biologic processes, medical diagnostics, etc., may be affected at lower concetrations of metallic-based nanocrystals made according to the techniques herein.
- the raw material metal ions used to grow the bi-metallic nanocrystals are provided by sacrificial metal electrodes used during the various electrochemical processes, there are no requirements for gold-based salts (or the equivalent) or platinum-based salts (or the equivalent) to be provided as raw materials for the formation of Au—Pt bi-metallic nanocrystal suspensions. Accordingly, components such as Cl ⁇ , chlorides or chlorine-based materials are not required to be part of the novel process or part of the novel bi-metallic nanocrystal suspensions produced. Additionally, no chlorine-based acids are required to produce the Au—Pt bi-metallic suspensions.
- metal-based bi-metallic nanocrystal suspensions or colloids of the present invention can be mixed or combined with other metallic-based solutions or colloids to form novel solution or colloid mixtures (e.g., in this instance, distinct metal species can still be discerned, etiher as composites or distinct species in a suspension).
- FIG. 1 shows a schematic cross-sectional view of a manual electrode assembly according to the present invention.
- FIG. 2 shows a schematic cross-sectional view of an automatic electrode control assembly according to the present invention.
- FIGS. 3 a - 3 e show five different representative embodiments of configurations for the electrode 1 .
- FIG. 4 shows a cross-sectional schematic view of plasmas produced utilizing one specific configuration of the electrode 1 corresponding to FIG. 3 e.
- FIGS. 5 a - 5 e show a variety of cross-sectional views of various trough members 30 .
- FIG. 6 shows a schematic cross-sectional view of a set of control devices 20 located on a trough member 30 with a liquid 3 flowing therethrough and into a storage container 41 .
- FIG. 7 a shows an AC transformer electrical wiring diagram for use with different embodiments of the invention.
- FIG. 7 b shows a schematic view of a transformer 60 and FIGS. 7 c and 7 d show schematic representations of two sine waves in phase and out of phase, respectively.
- FIG. 8 a shows a view of gold wires 5 a and 5 b used in some examples herein.
- FIG. 8 b shows a view of the gold wires 5 a and 5 b used in some examples herein.
- FIG. 8 c shows the device 20 used in all trough Examples herein that utilize a plasma.
- FIGS. 8 d , 8 e , 8 f and 8 g show wiring diagrams used to monitor and/or control the devices 20 .
- FIGS. 8 h and 8 i show wiring diagrams used to power devices 20 .
- FIG. 8 j shows a design for powering wires 5 / 5 in the devices 20 .
- FIG. 9 shows a first trough member 30 a ′ wherein one plasma 4 a is created. The output of this first trough member 30 a ′ flows into a second trough member 30 b′.
- FIGS. 10 a - 10 d show an alternative design of the trough member 30 b ′ wherein the trough member portions 30 a ′ and 30 b ′ are contiguous.
- FIGS. 11 a - 11 b show two trough members 30 b ′ used in connection with FIGS. 10 a - 10 d and various Examples herein.
- FIG. 11 c shows a representative TEM photomicrograph of dried gold constituents formed in connection with Example 1.
- FIG. 11 d shows a particle size distribution histogram from TEM measurements for the constituents formed in connection with Example 1.
- FIG. 11 e shows the UV-Vis spectral patterns of each of the gold suspension made according to Example 1.
- FIG. 12 a shows a schematic of an apparatus used in a batch method whereby in a first step, a plasma 4 is created to condition a fluid 3 ′.
- FIGS. 12 b and 12 c show a schematic of an apparatus used in a batch method utilizing wires 5 a and 5 b to form bi-metallic nanocrystals in suspension (e.g., a colloid) in association with the apparatus shown in FIG. 12 a and as discussed in various Examples herein.
- bi-metallic nanocrystals in suspension e.g., a colloid
- FIG. 12 d shows a schematic of an apparatus used in a batch method utilizing wires 5 a and 5 b to form bi-metallic nanocrystals in suspension (e.g., colloid) in association with the apparatus shown in FIG. 12 a , and as discussed in various examples herein.
- bi-metallic nanocrystals in suspension e.g., colloid
- FIG. 12 e shows a schematic view of the amplifier used in Examples 2 and 3.
- FIG. 12 f shows a schematic view of the power supply used in Examples 2 and 3.
- FIG. 12 g shows the UV-Vis spectral pattern of the Au—Pt bi-metallic suspensions made according to Example 6.
- FIG. 13 is a schematic of the power supply electrical setup used to generate the nanocrystals in the many Examples herein.
- FIG. 14 shows a representative TEM photomicrograph of dried platinum constituents formed in connection with Example 2.
- FIG. 15 a shows a representative TEM photomicrograph of dried platinum constituents formed in connection with Example 3.
- FIG. 15 b shows a particle size distribution histogram from TEM measurements for the constituents formed in connection with Example 3.
- FIG. 16 shows a representative TEM photomicrograph of dried platinum constituents formed in connection with Example 4.
- FIG. 17 shows the UV-Vis spectral patterns of each of the seven platinum solutions/suspensions made according to Example 5.
- FIG. 18 shows a representative TEM photomicrograph of the dried constituents made according to Example 6.
- FIG. 19 shows a representative TEM photomicrograph of the dried constituents made according to Example 7.
- FIG. 20 shows a representative TEM photomicrograph of the dried constituents made according to Example 8.
- FIGS. 21 a and 21 b show representative TEM photomicrographs of dried constituents made according to Example 9.
- FIGS. 22 a and 22 b are representative EDS spectra corresponding to FIGS. 21 a and 21 b , respectively.
- FIGS. 23 a and 23 b show representative TEM photomicrographs of dried constituents made according to Example 9.
- FIGS. 24 a and 24 b are representative EDS spectra corresponding to FIGS. 23 a and 23 b , respectively.
- FIG. 25 a shows a representative TEM photomicrograph of dried constituents made according to Example 10; and FIG. 25 b is a representative EDS spectra corresponding to FIG. 25 a.
- FIG. 26 a shows a representative TEM photomicrograph of dried constituents made according to Example 11; and FIG. 26 b is a representative EDS spectra corresponding to FIG. 26 a.
- FIG. 27 shows a UV-Vis spectrograph of GPB-032.
- FIG. 28 a shows three UV-Vis spectrographs of three Au—Pt bi-metallic suspensions.
- FIG. 28 b shows UV-Vis spectrographs for five different GPB bi-metallic suspensions.
- FIG. 28 c shows a graph of particle radius versus frequency for bi-metallic nanoparticles made according to Example 16.
- FIG. 29 a shows a representative TEM photomicrograph of the dried constituents made according to Example 17.
- FIG. 29 b is a representative EDS spectra corresponding to FIG. 29 a.
- FIG. 29 c shows a representative TEM photomicrograph of the dried constituents made according to Example 17.
- FIG. 29 d is a representative EDS spectra corresponding to FIG. 29 c.
- FIGS. 29 e , 29 f and 29 g are Scanning Transmission Electron Microscopy images of nanocrystals in a GPB-040 suspension.
- FIGS. 29 h and 29 i are representative XPS spectra corresponding to Example 17.
- FIG. 30 is a UV-Vis spectrograph of GPB-040 made according to Example 17.
- FIGS. 31 a and 31 b are schematic representations of the dialysis procedure used in Example 18; and FIG. 31 c is a schematic representation of a TFF apparatus.
- FIGS. 32 a - 32 ad are graphical depictions of anti-cancer activity of two suspensions (NE10214 and a bi-metallic nanocrystal suspension, GPB-032).
- FIGS. 33 a and 33 b show the results of the cancer xenograft tests set forth in Example 20a.
- FIGS. 34 a and 34 b show the results of the cancer xenograft tests set forth in Example 20b.
- FIGS. 35 a and 35 b show the results of the cancer xenograft tests set forth in Example 20c.
- FIGS. 36 a and 36 b show the results of the cancer xenograft tests set forth in Example 20d.
- FIGS. 37 a and 37 b show the results of the cancer xenograft tests set forth in Example 20e.
- FIGS. 38 a and 38 b show the results of the cancer xenograft tests set forth in Example 20f.
- FIGS. 39 a and 39 b represent the liquid consumption amount and weight gain for the mice set forth in Example 21.
- FIGS. 40 a and 40 b are graphs depicting the amount of absorbance of GPB-11 and various protein binders.
- FIG. 40 c shows an AFS photomicrograph of DNA binding to nanocrystals of GPB-11.
- New aqueous-based bi-metallic nanocrystal suspensions are manufactured from a combination of gold and platinum donor electrode materials, such bi-metallic nanocrystals including nanocrystalline surfaces that can be substantially free from organic or other impurities or films.
- the surfaces of the bi-metallic nanocrystals are “clean” relative to those surfaces of similar chemical composition nanoparticles made using: (1) chemical reduction processes that require chemical reductants and/or surfactants and/or various salt compounds as parts of the raw materials used to form bi-metallic-based nanoparticles from transition metal ions contained in raw material solution; and (2) other processes (including, sonoelectrochemistry, gamma-ray radiation, x-ray radiation, laser irradiation, electron accelorators, etc.) which use, for example, a variety of reductants or chlorine-based (or salt-based) raw materials (e.g., metal salts).
- the new bi-metallic nanocrystals of gold and platinum are produced via novel electrochemical manufacturing procedures, described in detail herein.
- the new electrochemical manufacturing procedures do not require the addition of chemical reductants and/or surfactants (e.g., organic compounds) or other agents, to be added to reduce metal ions and/or stabilize the formed bi-metallic nanocrystals.
- the processes do not require the addition of raw materials which contain both metal ions (which are reduced to form metal nanoparticles) and associated ions or species which counterbalance the electrical charge of the positively charged metal ion(s).
- Such added reductants, stabilizers and non-metal ion portions of raw materials are undesirable when they are typically carried along in, or on, the particles, or are undesirably adhered to at least a portion of the surface of the chemically reduced particles and/or remain as ions in the suspension. It is now understood that certain nanocrystal performance requirements can not be met with such impurities located on or bonded to the surface and such impurities need to be subsequently stripped or removed using various undesirable processes, which process themselves can affect the surface of the nanoparticles (e.g., plasma etching).
- a first set of electrochemical steps of the process involves the in situ creation of platinum species (e.g., raw materials) from a platinum metal source.
- the platinum species is created in water which contains a “process enhancer” or “processing enhancer” (typically an inorganic material or carbonate or such) which does not significantly bind to the formed nanocrystals in suspension, but rather facilitates removal of metal ions from a donor platinum metal electrode source, and/or assists in nucleation/growth during electrochemical-stimulated nanocrystal growth processes.
- the process enhancer serves important roles in the process including providing charged ions in the electrochemical solution to permit metal ions to be in solution and/or to cause the nanocrystals to be grown.
- the process enhancer is critically a compound(s) which remains in solution, and/or does not form a coating (e.g., an organic coating), and/or does not adversely affect the performance of the formed nanocrystals or the formed suspension(s) (e.g., is inert), and/or can be destroyed, evaporated, removed or otherwise lost during one or more steps of the electrochemical process.
- a preferred process enhancer is sodium bicarbonate. Examples of other process enhancers are sodium carbonate, sodium hydroxide, potassium bicarbonate, potassium carbonate, potassium hydroxide, trisodium phosphate, disodium phosphate, monosodium phosphate, potassium phosphates or the like and combinations thereof.
- Another particularly preferred processing enhancer is a mixture of sodium bicarbonate and potassium hydroxide.
- Desirable concentration ranges for the processing enhancer in the first step of the process include typically 0.01-20 grams/gallon (0.0026-2.1730 mg/ml), more typically, 0.1-7.5 grams/gallon (0.0264-1.9813 mg/ml) and most typically, 0.5-2.0 grams/gallon (0.13210-0.5283 mg/ml).
- desirable concentrations of the platinum species made in the first electrochemical steps of the process range from about 0.5 ppm to about 20 ppm and most typically about 1-8 ppm, and even more typically about 0.5-4 ppm.
- the result of the first set of electrochemical steps is a platform species in water.
- the platinum species can be predominantly nanocrystals or a mixture of nanocrystals and platinum ions.
- the platinum species is predominantly ions and the platinum ions—water material is used in a second set of electrochemical steps to form bi-metallic Au—Pt nanocrystals in suspension.
- a second set of steps of the electrochemical process involves the nucleation and growth of bi-metallic nanocrystals, such growth including: (1) mixtures of two metals, (2) alloys of two metals and/or (3) heteroaggregates (e.g., composites) of two metals.
- the platinum species and water output from the first steps of the preferred embodiment act as raw material input into the second electrochemical processing steps of a preferred embodiment.
- one or more of the aforementioned bi-metallic nanocrystalline components can be produced as stable nanocrystals in the aqueous suspension during the second set of electrochemical processing steps.
- bi-metallic nanocrystals have “bare” or “clean” surfaces of gold and/or platinum metal (e.g., in the zero oxidation state) bi-metallic nanocrystal surfaces are highly catalytic or are highly biocatalytic (as well as highly bioavailable).
- the bi-metallic nanocrystals are essentially surrounded by a water-based jacket comprising, for example, water species which are made available due to, for example, lysing of the water which occurs in one or more steps of a preferred embodiment.
- the lysed species may include hydrated electrons, OH ⁇ , H*, H 3 O, H 2 O 2 , etc.
- OH ⁇ groups may locate themselves around the formed bi-metallic crystals and create an electrostatic interaction therewith.
- These clean surface features provide novel and enhanced performance in a variety of industrial and medical applications and/or can result in decreased general undesirable toxicity in medical applications because no undesirable toxins or poisons are present on the surfaces due to the manufacturing process.
- the nanocrystals are not dried before use but instead are directly used in the liquid they were formed in (i.e., forming a suspension).
- the formed suspensions can be formed into a concentrate or a reconstituted concentrate thereof. It appears that completely removing these crystals from their suspension liquid (e.g., completely drying) may, in certain cases, adversely affect the surface properties of the crystals, (e.g., partial oxidation may occur, the stabilizing groups may be irreparably damaged, etc.) and/or may adversely affect the ability to rehydrate the crystals. For example, if the initially formed water jacket includes OH ⁇ which assist in electrostatic interactions, then changing the OH ⁇ coordination may upset the stability of the suspension.
- the dialysis procedure involves placement of the formed bi-metallic nanocrystal suspension inside of a dialysis bag.
- a polyethylene solution is located on the outside of the dialysis bag (e.g., the dialysis bag can be placed with a suitable container housing polyethylene glycol (PEG)) permits water to be removed from the formed bi-metallic nanocrystal suspension by osmotic pressure without comprising the stability of the nanocrystals in suspension.
- PEG polyethylene glycol
- sterile pharmaceutical grade water e.g., USP
- the water could be even more pure than USP by using reverse osmosis and/or ionic filtration means.
- the bi-metallic nanocrystals may be dried in situ into/onto, for example, an electrode or substrate which takes part in another reaction such as another electrochemical, chemical or catalytic process.
- the bi-metallic nanocrystals made according to this invention can also be used for industrial applications where metal reactivity is important (e.g., catalytic and/or electrochemical processes) but where pharmaceutical grade products/ingredients are not required.
- the bi-metallic nanocrystals can be made in a wider variety of solvents and with a wider variety of process enhancers, as discussed herein, depending on the specifc application. However, the clean aspects of the bi-metallic nanocrystal surfaces should be preserved to achieve superior performance.
- the electrochemical process steps of the invention can be controlled so as to result in more than one type of bi-metallic nanocrystal being present in the resultant suspension.
- mixtures of platinum and gold nanocrystals may exist in suspension
- alloys of platinum and gold nanocrystals may exist in suspension and/or nanocrystal heteroaggregates of platinum and gold may also exist in suspension.
- the bi-metallic nanocrystals can be grown in a manner that provides unique and identifiable surface characteristics such as spatially extended low index, crystal planes ⁇ 111 ⁇ , ⁇ 110 ⁇ and/or ⁇ 100 ⁇ and groups of such planes (and their equivalents). Such crystal planes can show different and desirable catalytic performances.
- a variety of crystalline shapes can be found in bi-metallic nanoparticle suspensions made according to embodiments disclosed herein. Further, the surfaces of bi-metallic nanocrystals grown should be highly active due to their crystalline condition (e.g., surface defects) as well as being clean.
- any desired average size of bi-metallic nanocrystals below 100 nm can be achieved.
- the most desirable nanocrystalline size ranges include those having an average crystal size (as measured and determined by specific techniques disclosed in detail herein) that is predominantly less than 100 nm, and more typically less than 50 nm, even more typically less than 30 nm, and in many of the preferred embodiments disclosed herein, the mode for the nanocrystal size distribution is less than 20 nm and within an even more preferable range of 8-18 nm.
- the techniques of the invention can be used to manufacture much larger particles.
- Resulting bi-metallic nanocrystalline suspensions or colloids can be provided that have or are adjusted to have target pH ranges.
- a sodium bicarbonate or other “basic” e.g., one where the OH ⁇ concentration is caused to be relatively high
- the pH range is typically 8-11, which can be adjusted as desired.
- certain processing enhancers can result in even higher pH ranges, such as a pH of about 9-12 or even 10.3-12.0.
- the nature and/or amount of the surface charge (i.e., positive or negative) on formed bi-metallic nanocrystals can have a large influence on the behavior and/or effects of the nanocrystal/suspension or colloid (or the concentrated nanocrystals).
- protein coronas such as albumin coronas and/or transferrin coronas formed in vivo can be influenced by surface charge or surface characteristics (e.g., including impurities or residual components present from processing techniques) of a nanoparticle. Such coronas dictate the biological identity of the nanoparticle and thus direct biologic availability.
- Such surface charges are commonly referred to as “zeta potential”. It is known that the larger the zeta potential (either positive or negative), the greater the stability of the nanoparticles in the solution (i.e., the suspension is more stable). By controlling the nature and/or amount of the surface charges of formed nanoparticles or nanocrystals, the performance of such nanoparticle suspensions can be controlled in biological and non-biological applications.
- Zeta potential is known as a measure of the electo-kinetic potential in colloidal systems and is also referred to as surface charge on particles.
- Zeta potential is the potential difference that exists between the stationary layer of fluid and the fluid within which the particle is dispersed.
- a zeta potential is often measured in millivolts (i.e., mV).
- the zeta potential value of approximately 20-25 mV is an arbitrary value that has been chosen to determine whether or not a dispersed particle is stable in a dispersion medium.
- zeta potential it should be understood that the zeta potential referred to is a description or quantification of the magnitude of the electrical charge present at the double layer.
- the zeta potential is calculated from the electrophoretic mobility by the Henry equation:
- ZP Zeta potentials
- the raw material metal ions are produced by the donor electrode metals of Pt and Au (e.g., sacrificial or donor electrodes) due to the processing conditions of the preferred embodiments.
- This “top-down” first set of electrochemical steps means that materials typically used to make metal-based nanoparticles in other techniques, such as metal salts (e.g., Pt salts, Au salts, etc.) are not required to be used in the embodiments disclosed herein.
- metal salts e.g., Pt salts, Au salts, etc.
- other constituents (which can be undesirable) of the metal salts such as Cl ⁇ or various chlorine-based materials, do not occur, or are not a required part of a product made according to the preferred embodiments herein.
- bi-metallic nanocrystal suspensions discussed herein can be chlorine or chloride-free.
- bi-metallic suspensions can be chlorine or chloride-free.
- chlorine-based materials dissolved in the suspension and were not required or essential to the nanoparticle production process, are contemplated as being within the metes and bounds of this disclosure.
- a set of novel process steps is provided to produce these unique bi-metallic nanocrystals.
- the process steps involve the creation of the bi-metallic nanocrystals in water.
- the water contains an added “process enhancer” which does not significantly bind to the formed nanocrystals, but rather facilitates nucleation/crystal growth during the electrochemical-stimulated growth process.
- the process enhancer serves important roles in the process including providing charged ions in the electrochemical solution to permit the crystals to be grown.
- These novel electrochemical processes can occur in either a batch, semi-continuous or continuous process. These processes result in controlled bi-metallic nanocrystalline concentrations of gold and platinum, controlled bi-metallic nanocrystal sizes and controlled bi-metallic nanocrystal size ranges.
- Novel manufacturing assemblies are provided to produce these bi-metallic nanocrystals.
- metallic-based constituents such as desirable metallic ions, can be included separately or combined with bi-metallic nanocrystal suspensions.
- the bi-metallic nanocrystal suspensions or colloids are made or grown by electrochemical techniques in either a batch, semi-continuous or continuous process, wherein the amount, average particle size, crystal plane(s) and/or particle shape(s) and/or particle shape distributions are controlled and/or optimized to achieve high biological activity and low cellular/biologic toxicity (e.g., a high therapeutic index).
- Desirable average crystal sizes include a variety of different ranges, but the most desirable ranges include average crystal sizes that are predominantly less than 100 nm and more typically, for many uses, less than 50 nm and even more typically for a variety of, for example, oral uses, less than 30 nm, and in many of the preferred embodiments disclosed herein, the mode for the nanocrystal size distribution is less than 20 nm and within an even more preferable range of 2-18 nm, as measured by a zetasizer (as described in more detail herein).
- the particles desirably contain crystal planes, such desirable (and often highly reactive) crystal planes, include crystals having ⁇ 111 ⁇ , ⁇ 110 ⁇ and/or ⁇ 100 ⁇ facets, as well as defects, which can result in superior interactions such as catalytic.
- these bi-metallic nanocrystals can be alloys, or can be combined with other metals in liquids such that metal “coatings” may occur on other metals to form composites or heteroaggregates or alternatively, mixtures of metal-based nanocrystals can be made.
- bi-metallic nanocrystal suspensions or colloids of the present invention can be mixed or combined with other metallic-based solutions or colloids to form novel solutions or colloid mixtures (e.g., in this instance, distinct metal species can still be discerned).
- Methods for making novel metallic-based nanocrystal suspensions or colloids relate generally to novel methods and novel devices for the continuous, semi-continuous and batch manufacture of a variety of constituents in a liquid including micron-sized particles, nanocrystals, ionic species and aqueous-based compositions of the same, including, nanocrystal/liquid(s), solution(s), colloid(s) or suspension(s).
- the constituents and bi-metallic nanocrystals produced can comprise a variety of possible compositions, concentrations, sizes, crystal planes (e.g., spatially extended low index crystal planes) and/or shapes, which together can cause the inventive compositions to exhibit a variety of novel and interesting physical, catalytic, biocatalytic and/or biophysical properties.
- the liquid(s) used and created/modified during the process can play an important role in the manufacturing of, and/or the functioning of the constituents (e.g., nanocrystals) independently or synergistically with the liquids which contain them.
- the particles (e.g., nanocrystals) are caused to be present (e.g., created and/or the liquid is predisposed to their presence (e.g., conditioned)) in at least one liquid (e.g., water) by, for example, typically utilizing at least one adjustable plasma (e.g., created by at least one AC and/or DC power source), which adjustable plasma communicates with at least a portion of a surface of the liquid.
- at least one liquid e.g., water
- adjustable plasma e.g., created by at least one AC and/or DC power source
- effective constituent (e.g., nanocrystals) suspensions or colloids can be achieved without the use of such plasmas as well.
- Gold and platinum-based electrodes of various composition(s) and/or unique configurations or arrangements are preferred for use in the formation of the adjustable plasma(s). Utilization of at least one subsequent and/or substantially simultaneous adjustable electrochemical processing technique is also preferred. Gold and platinum-based electrodes are preferred for use in the electrochemical processing technique(s). Electric fields, magnetic fields, electromagnetic fields, electrochemistry, pH, zeta potential, chemical/crystal constituents present, etc., are just some of the variables that can be positively affected by the adjustable plasma(s) and/or adjustable electrochemical processing technique(s) of the invention.
- adjustable plasmas and/or adjustable electrochemical techniques are preferred in many embodiments of the invention to achieve many of the processing advantages of the present invention, as well as many of the novel bi-metallic nanocrystals and bi-metallic nanocrystal compositions which result from practicing the teachings of the preferred embodiments to make an almost limitless set of inventive aqueous solutions, suspensions and/or colloids.
- At least one liquid flows into, through and out of at least one first trough member and such liquid is processed, conditioned, modified and/or effected by said at least one adjustable plasma and/or said at least one adjustable electrochemical technique.
- the results of the continuous processing in the first trough member include new constituents in the liquid, such as ionic constituents, nanocrystals (e.g., platinum-based nanocrystals) of novel and/or controllable size, hydrodynamic radius, concentration, crystal sizes and crystal size ranges, zeta potential, pH and/or properties, such platinum nanocrystal/ion/liquid mixture being produced in an efficient and economical manner.
- a first set of steps of the process involves the in situ creation of platinum species (e.g., raw materials) from a platinum metal source.
- the platinum species is created in water which contains a “process enhancer” or “processing enhancer” (typically an inorganic material or carbonate or such) which does not significantly bind to the formed nanocrystals in suspension, but rather facilitates removal of metal ions from a donor metal source, and/or assists in nucleation/growth during electrochemical-stimulated nanocrystal growth processes.
- the process enhancer serves important roles in the process including providing charged ions in the electrochemical solution to permit the nanocrystals to be grown.
- the process enhancer is critically a compound(s) which remains in solution, and/or does not form a coating (e.g., an organic coating), and/or does not adversely affect the performance of the formed nanocrystals or the formed suspension(s) (e.g., is inert), and/or can be destroyed, evaporated, removed or otherwise lost during one or more steps of the electrochemical process.
- a preferred process enhancer is sodium bicarbonate. Examples of other process enhancers are sodium carbonate, potassium bicarbonate, potassium carbonate, trisodium phosphate, disodium phosphate, monosodium phosphate, potassium phosphates or the like and combinations thereof.
- Another particularly preferred processing enhancer is a mixture of sodium bicarbonate and potassium hydroxide.
- Desirable concentration ranges for the processing enhancer include typically 0.01-20 grams/gallon (0.0026-2.1730 mg/ml), more typically, 0.1-7.5 grams/gallon (0.0264-1.9813 mg/ml) and most typically, 0.5-2.0 grams/gallon (0.13210-0.5283 mg/ml).
- a second set of steps of the process involves the nucleation and growth of bi-metallic-based nanocrystals, such growth being: (1) mixtures of two metals, (2) alloys of two metals and/or (3) heteroaggregates of two metals.
- the aqueous output from the first steps of the preferred embodiment containing water, platinum species resulting from the first steps of the process, and processing enhancer used during the first set of steps acts as raw material input into the second electrochemical steps of a preferred embodiment.
- one or more of the aforementioned bi-metallic nanocrystalline components can be produced as stable bi-metallic nanocrystals in the aqueous suspension during the second set of steps.
- Certain processing enhancers may dissociate into positive ions (cations) and negative ions (anions).
- the anions and/or cations depending on a variety of factors including liquid composition, concentration of ions, change state of ions, applied fields, frequency of applied fields, waveform of the applied filed, temperature, pH, zeta potential, etc., will navigate or move toward oppositely charged electrodes.
- the ions When said ions are located at or near such electrodes, the ions may take part in one or more reactions with the electrode(s) and/or other constituent(s) located or created at or near such electrode(s). Sometimes ions may react with one or more materials in the electrode. Such reactions may be desirable in some cases or undesirable in others.
- ions present in a solution between electrodes may not react to form a product, but rather may influence material in the electrode (or near the electrode) to form metallic nano-crystals that are “grown” from material provided by the donor electrode.
- certain metal ions may enter the liquid 3 from the electrode 5 and be caused to come together (e.g., nucleate) to form constituents (e.g., ions, nanocrystals, etc.) within the liquid 3 .
- a process enhancer that will not negatively impact performance such as, for example, impart negative performance or, for example, toxicity to the bi-metallic nanocrystal, or to the liquid that the crystal is suspended in, to maximize acceptability for various commercial uses (e.g., pharmaceutical, catalytic, medical diagnostic, etc).
- chlorine ions or chlorides or chlorine-based materials may be undesired if such species create, for example, gold chloride salts, which may be undesirable for several reasons (e.g., may affect toxicity, stability, etc.).
- processing enhancers that involve hydroxyl groups OH ⁇ (e.g., which are part of the processing enhancer or result from addition of processing enhancers to the liquid 3 ) can also be desirable.
- desirable processing enhancers of NaOH, KOH and NaHCO 3 (and mixtures of the same) are specifically disclosed as being desirable in some preferred embodiments herein.
- drying, concentrating and/or freeze drying can also be utilized to remove at least a portion of, or substantially all of, the suspending liquid, resulting in, for example, partially or substantially completely dehydrated bi-metallic nanocrystals. If such nanocrystals are ultimately located on a substrate (e.g., a catalysis substrate or an electrode) complete drying may be required. If solutions, suspensions or colloids are completely dehydrated, the metal-based species, in some cases, should be capable of being rehydrated by the addition of liquid (e.g., of similar or different composition than that which was removed). However, not all compositions/colloids of the present invention can be completely dehydrated without adversely affecting performance of the composition/colloid.
- nanocrystals formed in a liquid tend to clump or stick together (or adhere to surfaces) when dried. If such clumping is not reversible during a subsequent rehydration step, dehydration should be avoided. However, for a variety of applications such clumping may be acceptable. Further, when drying on a substrate, such clumping may be avoided.
- the dialysis procedure involves placement of the formed bi-metallic nanocrystal suspension inside of a dialysis bag.
- a polyethylene solution is located on the outside of the dialysis bag (e.g., the dialysis bag can be placed with a suitable container holding polyethylene glycol (PEG)) and water can be removed from the formed bi-metallic nanocrystal suspension by osmotic pressure without comprising the stability of the nanocrystals in suspension.
- PEG polyethylene glycol
- adjustable plasma which adjustable plasma is located between at least one electrode positioned adjacent to (e.g., above) at least a portion of the surface of a liquid (e.g., water) and at least a portion of the surface of the liquid itself.
- the liquid is placed into electrical communication with at least one second electrode (or a plurality of second electrodes) causing the surface of the liquid to function as an electrode, thus taking part in the formation of the adjustable plasma.
- This configuration has certain characteristics similar to a dielectric barrier discharge configuration, except that the surface of the liquid is an active electrode participant in this configuration.
- Each adjustable plasma utilized can be located between the at least one electrode located above a surface of the liquid and a surface of the liquid due to at least one electrically conductive electrode being located somewhere within (e.g., at least partially within) the liquid.
- At least one power source in a preferred embodiment, at least one source of volts and amps such as a transformer or power source is connected electrically between the at least one electrode located above the surface of the liquid and the at least one electrode contacting the surface of the liquid (e.g., located at least partially, or substantially completely, within the liquid).
- the electrode(s) may be of any suitable composition (however, platinum and gold are preferred) and suitable physical configuration (e.g., size and shape) which results in the creation of a desirable plasma between the electrode(s) located above the surface of the liquid and at least a portion of the surface of the liquid itself.
- the applied power (e.g., voltage and amperage) between the electrode(s) (e.g., including the surface of the liquid functioning as at least one electrode for forming the plasma) can be generated by any suitable source (e.g., voltage from a transformer) including both AC and DC sources and variants and combinations thereof.
- the electrode or electrode combination located within e.g., at least partially below the surface of the liquid
- the adjustable plasma is actually located between at least a portion of the electrode(s) located above the surface of the liquid (e.g., at a tip or point thereof) and one or more portions or areas of the liquid surface itself.
- the adjustable plasma can be created between the aforementioned electrodes (i.e., those located above at least a portion of the surface of the liquid and a portion of the liquid surface itself) when a breakdown voltage of the gas or vapor around and/or between the electrode(s) and the surface of the liquid is achieved or maintained.
- the liquid comprises water (or water containing certain processing enhancer(s)), and the gas between the surface of the water and the electrode(s) above the surface of the water (i.e., that gas or atmosphere that takes part in the formation of the adjustable plasma) comprises air.
- the air can be controlled to contain various different water content(s) or a desired humidity which can result in different compositions, concentrations, crystal size distributions and/or crystal shape distributions of constituents (e.g., nanocrystals) being produced according to the present invention (e.g., different amounts of certain constituents in the adjustable plasma and/or in the solution or suspension can be a function of the water content in the air located above the surface of the liquid) as well as different processing times required to obtain certain concentrations of various constituents in the liquid, etc.
- constituents e.g., nanocrystals
- Equation (1) gives the empirical relationship between the breakdown electric field “E c ” and the distance “d” (in meters) between two electrodes:
- the breakdown electric field “E c ” will vary as a function of the properties and composition of the gas or vapor located between electrodes.
- water or water containing a processing enhancer
- significant amounts of water vapor can be inherently present in the air between the “electrodes” (i.e., between the at least one electrode located above the surface of the water and the water surface itself which is functioning as one electrode for plasma formation) and such water vapor should have an effect on at least the breakdown electric field required to create a plasma therebetween.
- a higher concentration of water vapor can be caused to be present locally in and around the created plasma due to the interaction of the adjustable plasma with the surface of the water.
- the amount of “humidity” present in and around the created plasma can be controlled or adjusted by a variety of techniques discussed in greater detail later herein.
- certain components present in any liquid can form at least a portion of the constituents forming the adjustable plasma located between the surface of the liquid and the electrode(s) located adjacent (e.g., along) the surface of the liquid.
- the constituents in the adjustable plasma, as well as the physical properties of the plasma per se, can have a dramatic influence on the liquid, as well as on certain of the processing techniques (discussed in greater detail later herein).
- the electric field strengths created at and near the electrodes are typically at a maximum at a surface of an electrode and typically decrease with increasing distance therefrom.
- a portion of the volume of gas between the electrode(s) located above a surface of a liquid and at least a portion of the liquid surface itself can contain a sufficient breakdown electric field to create the adjustable plasma.
- These created electric fields can influence, for example, behavior of the adjustable plasma, behavior of the liquid (e.g., influence the crystal state of the liquid) behavior of constituents in the liquid, etc.
- FIG. 1 shows one embodiment of a point source electrode 1 having a triangular cross-sectional shape located a distance “x” above the surface 2 of a liquid 3 flowing, for example, in the direction “F”.
- An adjustable plasma 4 can be generated between the tip or point 9 of the electrode 1 and the surface 2 of the liquid 3 when an appropriate power source 10 is connected between the point source electrode 1 and the electrode 5 , which electrode 5 communicates with the liquid 3 (e.g., is at least partially below the surface 2 of the liquid 3 ).
- the adjustable plasma region 4 created in the embodiment shown in FIG. 1 can typically have a shape corresponding to a cone-like structure or an ellipsoid-like structure, for at least a portion of the process, and in some embodiments of the invention, can maintain such shape (e.g., cone-like shape) for substantially all of the process.
- the volume, intensity, constituents (e.g., composition), activity, precise locations, etc., of the adjustable plasma(s) 4 will vary depending on a number of factors including, but not limited to, the distance “x”, the physical and/or chemical composition of the electrode 1 , the shape of the electrode 1 , the power source 10 (e.g., DC, AC, rectified AC, the applied polarity of DC and/or rectified AC, AC or DC waveform, RF, etc.), the power applied by the power source (e.g., the volts applied, which is typically 1000-5000 Volts, and more typically 1000-1500 Volts, the amps applied, electron velocity, etc.) the frequency and/or magnitude of the electric and/or magnetic fields created by the power source applied or ambient, electric, magnetic or electromagnetic fields, acoustic fields, the composition of the naturally occurring or supplied gas or atmosphere (e.g., air, nitrogen, helium, oxygen, ozone, reducing atmospheres, etc.) between and/or around the electrode 1 and the surface
- the composition of the electrode(s) 1 involved in the creation of the adjustable plasma(s) 4 of FIG. 1 are metal-based compositions (e.g., metals such as gold, platinum and/or alloys or mixtures thereof, etc.), but the electrodes 1 and 5 may be made out of any suitable material compatible with the various aspects (e.g., processing parameters) of the inventions disclosed herein.
- a plasma 4 in, for example, air above the surface 2 of a liquid 3 (e.g., water) will, typically, produce at least some ozone, as well as amounts of nitrogen oxide and other components.
- the adjustable plasma 4 actually contacts the surface 2 of the liquid 3 .
- material e.g., metal
- material from the electrode 1 may comprise a portion of the adjustable plasma 4 (e.g., and thus be part of the emission spectrum of the plasma) and may be caused, for example, to be “sputtered” onto and/or into the liquid 3 (e.g., water).
- the electrode(s) 1 when metal(s) are used as the electrode(s) 1 , a variety of constituents can be formed in the electrical plasma, resulting in certain constituents becoming part of the processing liquid 3 (e.g., water), including, but not limited to, elementary metal(s), metal ions, Lewis acids, Bronsted-Lowry acids, metal oxides, metal nitrides, metal hydrides, metal hydrates and/or metal carbides, etc., can be found in the liquid 3 (e.g., for at least a portion of the process and may be capable of being involved in simultaneous/subsequent reactions), depending upon the particular set of operating conditions associated with the adjustable plasma 4 and/or subsequent electrochemical processing operations.
- the processing liquid 3 e.g., water
- elementary metal(s) e.g., metal ions, Lewis acids, Bronsted-Lowry acids, metal oxides, metal nitrides, metal hydrides, metal hydrates and/or metal carbides, etc.
- Such constituents may be transiently present in the processing liquid 3 or may be semi-permanent or permanent. If such constituents are transient or semi-permanent, then the timing of subsequent reactions (e.g., electrochemical reactions) with such formed constituents can influence final products produced. If such constituents are permanent, they should not adversely affect the desired performance of the active ingredient nanocrystals.
- electrode(s) 1 and 5 the physical and chemical construction of the electrode(s) 1 and 5 , atmosphere (naturally occurring or supplied), liquid composition, greater or lesser amounts of electrode(s) materials(s) (e.g., metal(s) or derivatives of metals) may be found in the liquid 3 .
- electrode(s) materials(s) e.g., metal(s) or derivatives of metals
- the material(s) e.g., metal(s) or metal(s) composite(s)
- constituents e.g., Lewis acids, Bronsted-Lowry acids, etc.
- the material(s) e.g., metal(s) or metal(s) composite(s)
- constituents e.g., Lewis acids, Bronsted-Lowry acids, etc.
- electrode composition can play an important role in the materials that are formed according to the embodiments disclosed herein. The interplay between these components of the invention are discussed in greater detail later herein.
- the electrode(s) 1 and 5 may be of similar chemical composition (e.g., have the same chemical element as their primary constituent) and/or mechanical configuration or completely different compositions (e.g., have different chemical elements as their primary constituent) in order to achieve various compositions and/or structures of liquids and/or specific effects discussed later herein.
- the distance “y” between the electrode(s) 1 and 5 ; or 1 and 1 (shown later herein) or 5 and 5 (shown later herein) is one important aspect of the invention.
- the location of the smallest distance “y” between the closest portions of the electrode(s) used in the present invention should be greater than the distance “x” in order to prevent an undesirable arc or formation of an unwanted corona or plasma occurring between the electrode (e.g., the electrode(s) 1 and the electrode(s) 5 ) (unless some type of electrical insulation is provided therebetween).
- the power applied through the power source 10 may be any suitable power which creates a desirable adjustable plasma 4 under all of the process conditions of the present invention.
- an alternating current from a step-up transformer is utilized.
- Preferred transformer(s) 60 (see e.g., FIGS. 7 a - 7 b ) for use in various embodiments disclosed herein, have deliberately poor output voltage regulation made possible by the use of magnetic shunts in the transformer 60 .
- These transformers 60 are known as neon sign transformers. This configuration limits current flow into the electrode(s) 1 / 5 . With a large change in output load voltage, the transformer 60 maintains output load current within a relatively narrow range.
- the transformer 60 is rated for its secondary open circuit voltage and secondary short circuit current.
- Open circuit voltage (OCV) appears at the output terminals of the transformer 60 only when no electrical connection is present.
- short circuit current is only drawn from the output terminals if a short is placed across those terminals (in which case the output voltage equals zero).
- OCV Open circuit voltage
- the output voltage of the transformer 60 should fall somewhere between zero and the rated OCV. In fact, if the transformer 60 is loaded properly, that voltage will be about half the rated OCV.
- the transformer 60 is known as a Balanced Mid-Point Referenced Design (e.g., also formerly known as balanced midpoint grounded). This is most commonly found in mid to higher voltage rated transformers and most 60 mA transformers. This is the only type transformer acceptable in a “mid-point return wired” system.
- the “balanced” transformer 60 has one primary coil 601 with two secondary coils 603 , one on each side of the primary coil 601 (as shown generally in the schematic view in FIG. 7 b ). This transformer 60 can in many ways perform like two transformers.
- each secondary coil 603 is attached to the core 602 and subsequently to the transformer enclosure and the other end of the each secondary coil 603 is attached to an output lead or terminal.
- an unloaded 15,000 volt transformer of this type will measure about 7,500 volts from each secondary terminal to the transformer enclosure but will measure about 15,000 volts between the two output terminals.
- These exemplary transformers 60 were utilized to form the plasmas 4 disclosed in the Examples herein. However, other suitable transformers (or power sources) should also be understood as falling within the metes and bounds of the invention. However, a different power supply 501 AC (discussed elsewhere herein) is utilized for the electrodes 5 / 5 ′ in most of the other examples disclosed herein.
- electrode holders 6 a and 6 b are capable of being lowered and raised by any suitable means (and thus the electrodes are capable of being lowered and raised).
- the electrode holders 6 a and 6 b are capable of being lowered and raised in and through an insulating member 8 (shown in cross-section).
- the mechanical embodiment shown here includes male/female screw threads.
- the portions 6 a and 6 b can be covered by, for example, additional electrical insulating portions 7 a and 7 b .
- the electrical insulating portions 7 a and 7 b can be any suitable material (e.g., plastic, polycarbonate, poly (methyl methacrylate), polystyrene, acrylics, polyvinylchloride (PVC), nylon, rubber, fibrous materials, etc.) which prevent undesirable currents, voltage, arcing, etc., that could occur when an individual interfaces with the electrode holders 6 a and 6 b (e.g., attempts to adjust the height of the electrodes).
- the insulating member 8 can be made of any suitable material which prevents undesirable electrical events (e.g., arcing, melting, etc.) from occurring, as well as any material which is structurally and environmentally suitable for practicing the present invention. Typical materials include structural plastics such as polycarbonates, plexiglass (poly (methyl methacrylate), polystyrene, acrylics, and the like. Additional suitable materials for use with the present invention are discussed in greater detail elsewhere herein.
- the power source 10 can be connected in any convenient electrical manner to the electrodes 1 and 5 .
- wires 11 a and 11 b can be located within at least a portion of the electrode holders 6 a , 6 b (and/or electrical insulating portions 7 a , 7 b ) with a primary goal being achieving electrical connections between the portions 11 a , 11 b and thus the electrodes 1 , 5 .
- FIG. 2 shows another schematic of a preferred embodiment of the invention, wherein a control device 20 is connected to the electrodes 1 and 5 , such that the control device 20 remotely (e.g., upon command from another device or component) raises and/or lowers the electrodes 1 , 5 relative to the surface 2 of the liquid 3 .
- the control device 20 is discussed in more detail later herein.
- the electrodes 1 and 5 can be, for example, remotely lowered and controlled, and can also be monitored and controlled by a suitable controller or computer (not shown in FIG. 2 ) containing an appropriate software control program. Accordingly, the embodiment shown in FIG. 1 should be considered to be a manually controlled apparatus for use with the techniques of the present invention, whereas the embodiment shown in FIG.
- FIG. 2 should be considered to include an automatic apparatus or assembly 20 which can remotely raise and lower the electrodes 1 and 5 in response to appropriate commands.
- the FIG. 2 preferred embodiments of the invention can also employ computer monitoring and computer control of the distance “x” of the tips 9 of the electrodes 1 (and tips 9 ′ of the electrodes 5 ) away from the surface 2 ; or computer monitoring and/or controlling the rate(s) which the electrode 5 is advanced into/through the liquid 3 .
- the appropriate commands for raising and/or lowering the electrodes 1 and 5 can come from an individual operator and/or a suitable control device such as a controller or a computer (not shown in FIG. 2 ).
- FIGS. 3 a - 3 e show perspective views of various desirable electrode configurations for the electrode 1 shown in FIGS. 1 - 2 (as well as in other Figures and embodiments discussed later herein).
- the electrode configurations shown in FIGS. 3 a - 3 e are representative of a number of different configurations that are useful in various embodiments of the present invention.
- Criteria for appropriate electrode selection for the electrode 1 include, but are not limited to the following conditions: the need for a very well defined tip or point 9 , composition, mechanical limitations, the ability to make shapes from the material comprising the electrode 1 , conditioning (e.g., heat treating or annealing) of the material comprising the electrode 1 , convenience, the constituents introduced into the plasma 4 , the influence upon the liquid 3 , etc.
- a small mass of material comprising the electrodes 1 shown in, for example, FIGS. 1 - 2 may, upon creation of the adjustable plasmas 4 according to the present invention (discussed in greater detail later herein), rise to operating temperatures where the size and or shape of the electrode(s) 1 can be adversely affected.
- the electrode 1 was of relatively small mass (e.g., if the electrode(s) 1 was made of gold and weighed about 0.5 gram or less) and included a very fine point as the tip 9 , then it is possible that under certain sets of conditions used in various embodiments herein that a fine point (e.g., a thin wire having a diameter of only a few millimeters and exposed to a few hundred to a few thousand volts; or a triangular-shaped piece of metal) would be incapable of functioning as the electrode 1 (e.g., the electrode 1 could deform undesirably or melt), absent some type of additional interactions (e.g., internal cooling means or external cooling means such as a fan, etc.).
- a fine point e.g., a thin wire having a diameter of only a few millimeters and exposed to a few hundred to a few thousand volts; or a triangular-shaped piece of metal
- the electrode 1 could deform undesirably or melt
- composition of (e.g., the material comprising) the electrode(s) 1 may affect possible suitable electrode physical shape due to, for example, melting points, pressure sensitivities, environmental reactions (e.g., the local environment of the adjustable plasma 4 could cause undesirable chemical, mechanical and/or electrochemical erosion of the electrode(s)), etc.
- the electrode 1 shown in FIG. 3 e comprises a rounded tip 9 .
- partially rounded or arc-shaped electrodes can also function as the electrode 1 because the adjustable plasma 4 , which is created in the inventive embodiments shown herein (see, for example, FIGS. 1 - 2 ), can be created from rounded electrodes or electrodes with sharper or more pointed features.
- such adjustable plasmas can be positioned or can be located along various points of the electrode 1 shown in FIG. 3 e . In this regard, FIG.
- Electrode 4 shows a variety of points “a-g” which correspond to initiating points 9 for the plasmas 4 a - 4 g which occur between the electrode 1 and the surface 2 of the liquid 3 . Accordingly, it should be understood that a variety of sizes and shapes corresponding to electrode 1 can be utilized in accordance with the teachings of the present invention. Still further, it should be noted that the tips 9 , 9 ′ of the electrodes 1 and 5 , respectively, shown in various Figures herein, may be shown as a relatively sharp point or a relatively blunt end. Unless specific aspects of these electrode tips 9 , 9 ′ are discussed in greater contextual detail, the actual shape of the electrode tip(s) 9 , 9 ′ shown in the Figures should not be given great significance.
- the electrode configurations shown generally in FIGS. 1 and 2 can create different results (e.g., different conditioning effects for the fluid 3 , different pH's in the fluid 3 , different nanocrystals sizes and size distribution, different nanocrystal shapes and nanocrystal shape distributions, and/or amounts of constituents (e.g., nanocrystal matter and/or metal ions from the donor electrode(s)) found in the fluid 3 , different functioning of the fluid/nanocrystal combinations (e.g., different biologic/biocatalytic effects), different zeta potentials, etc.) as a function of a variety of features including the electrode orientation and position relative to the fluid flow direction “F”, cross-sectional shape and size of the trough member 30 (or 30 a ′ and/or 30 b ′), and/or amount of the liquid 3 within the trough member 30 and/or rate of flow of the liquid 3 within the trough member 30 and in/around the electrodes 5 a / 5 b , the thickness of the electrode
- the electrode compositions, size, specific shape(s), number of different types of electrodes provided, voltage applied, amperage applied and/or achieved within the liquid 3 , AC source (and AC source frequency and AC waveform shape, duty cycle, etc.), DC source, RF source (and RF source frequency, duty cycle, etc.), electrode polarity, etc. can all influence the properties of the liquid 3 (and/or the nanocrystals formed or contained in the liquid 3 ) as the liquid 3 contacts, interacts with and/or flows past these electrodes 1 , 5 and hence resultant properties of the materials (e.g., the nanocrystals produced, metal ions, and/or the suspension or colloid) produced therefrom.
- FIGS. 5 a - 5 e show cross-sectional views of the liquid containing trough member 30 used in preferred embodiments herein.
- the distance “S” and “S′” for the preferred embodiment shown in each of FIGS. 5 a - 5 e measures, for example, between about 0.25′′ and about 6′′ (about 0.6 cm-15 cm).
- the distance “M” ranges from about 0.25′′ to about 6′′ (about 0.6 cm-15 cm).
- the distance “R” ranges from about 1 ⁇ 2′′ to about 7′′ (about 1.2 cm to about 17.8 cm). All of these embodiments (as well as additional configurations that represent alternative embodiments are within the metes and bounds of this inventive disclosure) can be utilized in combination with the other inventive aspects of the invention.
- the amount of liquid 3 contained within each of the liquid containing trough members 30 is a function not only of the depth “d”, but also a function of the actual cross-section.
- the amount of liquid 3 present in and around the electrode(s) 1 and 5 can influence one or more effects of the adjustable plasma 4 upon the liquid 3 as well as the electrochemical interaction(s) of the electrode 5 with the liquid 3 .
- the flow rate of the liquid 3 in and around the electrode(s) 1 and 5 can also influence many of properties of the nanocrystals formed in the resulting colloids or suspensions.
- adjustable plasma 4 conditioning effects e.g., interactions of the plasma electric and magnetic fields, interactions of the electromagnetic radiation of the plasma, creation of various chemical species (e.g., Lewis acids, Bronsted-Lowry acids) within the liquid, pH changes, temperature variations of the liquid (e.g., slower liquid flow can result in higher liquid temperatures and/or longer contact or dwell time with or around the electrodes 1 / 5 which can also desirably influence final products produced, such as size/shape of the formed nanocrystals, etc.) upon the liquid 3 , but also the concentration or interaction of the adjustable plasma 4 with the liquid 3 .
- adjustable plasma 4 conditioning effects e.g., interactions of the plasma electric and magnetic fields, interactions of the electromagnetic radiation of the plasma, creation of various chemical species (e.g., Lewis acids, Bronsted-Lowry acids) within the liquid, pH changes, temperature variations of the liquid (e.g., slower liquid flow can result in higher liquid temperatures and/or longer contact or dwell time with or around the electrodes 1 / 5 which can also des
- the influence of many aspects of the electrode 5 on the liquid 3 is also, at least partially, a function of the amount of liquid juxtaposed to the electrode(s) 5 . All of these factors can influence a balance which exists between nucleation and growth of the nanocrystals grown in the liquid 3 , resulting in, for example, particle size and size range control and/or particle shape and shape range control.
- the initial temperature of the liquid 3 input into the trough member 30 can also affect a variety of properties of products produced according to the disclosure herein.
- different temperatures of the liquid 3 can affect nanocrystal size(s) and nanocrystal shape(s), concentration or amounts of various formed constituents (e.g., transient, semi-permanent or permanent constituents), ionic control of the liquid, pH, zeta potential, etc.
- temperature controls along at least a portion of, or substantially all of, the trough member 30 (or 30 a ′ and/or 30 b ′) can have desirable effects.
- resultant properties of products formed can be controlled.
- Preferable liquid 3 temperatures during the processing thereof are between freezing and boiling points, more typically, between room temperature and boiling points, and even more typically, between about 40-98 degrees C., and more typically, between about 50-98 degrees C.
- Such temperature can be controlled by, for example, conventional means for cooling located at or near various portions of the processing apparatus.
- processing enhancers may also be added to or mixed with the liquid(s) 3 .
- the processing enhancers include both solids and liquids (and gases in some cases).
- the processing enhancer(s) may provide certain processing advantages and/or desirable final product characteristics. Some portion of the processing enhancer(s) may function, influence as or become part of, for example, desirable seed crystals (or promote desirable seed crystals, or be involved in the creation of a nucleation site) and/or crystal plane growth promoters/preventers in the electrochemical growth processes of the invention; or may simply function as a current or power regulator in the electrochemical processes of the invention. Such processing enhancers may also desirably affect current and/or voltage conditions between electrodes 1 / 5 and/or 5 / 5 .
- a preferred processing enhancer is sodium bicarbonate.
- examples of other process enhancers are sodium carbonate, potassium bicarbonate, potassium carbonate, trisodium phosphate, disodium phosphate, monosodium phosphate, potassium hydroxide, potassium phosphates or the like and combinations thereof.
- Another particularly preferred processing enhancer is a mixture of sodium bicarbonate and potassium hydroxide.
- Still other process enhancers to make bi-metallic nanocrystals for medical applications under certain conditions may be any material that assists in the electrochemical growth processes described herein; and any material is not substantially incorporated into or onto the surface of the gold nanocrystasl; and does not impart toxicity to the nanocrystals or to the suspension containing the nanocrystals.
- Processing enhancers may assist in one or more of the electrochemical reactions disclosed herein; and/or may assist in achieving one or more desirable properties in products formed according to the teachings herein.
- processing enhancers do not contain Cl ⁇ or chlorides or chlorine-based materials which are required by other processing techniques.
- certain processing enhancers may dissociate into positive ions (cations) and negative ions (anions).
- the anions and/or cations depending on a variety of factors including liquid composition, concentration of ions, applied fields, frequency of applied fields, waveform of the applied filed, temperature, pH, zeta potential, etc., will navigate or move toward oppositely charged electrodes.
- the ions When said ions are located at or near such electrodes, the ions may take part in one or more intermediate reactions with the electrode(s) and/or other constituent(s) located at or near such electrode(s). Sometimes ions may react with one or more materials in the electrode and cause metallic ions to be produced in the liquid.
- ions present in a solution between electrodes may influence material in the electrode (or near the electrode) to form metallic nano-crystals that are “grown” from material provided by the electrode.
- certain metal ions may enter the liquid 3 from the electrode 5 and be caused to come together (e.g., nucleate) to form constituents (e.g., ions, nanocrystals, etc.) within the liquid 3 .
- Such ions can then be used as a raw material for the growth of bi-metallic nanocrystals.
- nanocrystalline shapes or shape distributions
- specific spatially extended low index crystal planes can cause different reactions (e.g., different catalytic, electrochemical, biocatalytic and/or biophysical reactions and/or cause different biological signaling pathways to be active/inactive relative to the absence of such shaped nanoparticles) and/or different reactions selectively to occur under substantially identical conditions.
- Such differences in performance may be due to differing surface plasmon resonances and/or intensity of such resonances.
- certain reactions e.g., catalytic, electrochemical, biological reactions and/or biological signaling pathways
- control can result in the prevention and/or treatment of a variety of different diseases or indications that are a function of certain biologic reactions and/or signaling pathways, as well as control of a number of non-biological reaction pathways.
- processing enhancers may also include materials that may function as charge carriers, but may themselves not be ions.
- metallic-based particles either introduced or formed in situ (e.g., heterogeneous or homogenous nucleation/growth) by the electrochemical processing techniques disclosed herein, can also function as charge carriers, crystal nucleators and/or growth promoters, which may result in the formation of a variety of different crystalline shapes (e.g., hexagonal plates, octahedrons, techahedrons, pentagonal bi-pyramids (decahedrons), etc.).
- the presence of particular particle crystal sizes, extended crystal planes and/or shapes or shape distributions of such crystals can desirably influence certain reactions (e.g., binding to a particular protein or protein homologue and/or affecting a particular biological signaling pathway such as an inflammatory pathway or a proteasomal pathway) to occur.
- certain reactions e.g., binding to a particular protein or protein homologue and/or affecting a particular biological signaling pathway such as an inflammatory pathway or a proteasomal pathway
- platinum species that are formed in a first trough member 30 a ′/ 30 b ′ are caused to flow into a second trough member 30 a ′/ 30 b ′ and take part in the formation of bi-metallic nanocrystals therein. More specifically, a first set of electrochemical reactions occur in a water containing a suitable processing enhancer to create a modified water-processing enhancer solution/suspension, which then serves as a raw material supply for a second set of electrochemical reactions that occur in a second trough member 30 a ′/ 30 b ′.
- the two separate trough members are kept as separate members and the output of the first trough member is allowed to cool before being input into the second trough member.
- the two trough members can be an integral unit, with or without cooling means located between the two identifiable portions 30 a ′/ 30 b′.
- the processing enhancers of the present invention do not contemplate those traditional organic-based molecules used in traditional reduction chemistry techniques, the lack of such chemical reductant (or added surfactant) means that the surfaces of the grown nanocrystals on the invention are very “clean” relative to nanoparticles that are formed by traditional reduction chemistry approaches.
- nanocrystal surfaces when the term “clean” is used with regard to nanocrystal surfaces or when the phrase “substantially free from organic impurities or films” (or a similar phrase) is used, what is meant is that the formed nanocrystals do not have chemical constituents adhered or attached to their surfaces which (1) alter the functioning of the nanocrystal and/or (2) form a layer, surface or film which covers a significant portion (e.g., at least 25% of the crystal, or more typically, at least 50% of the crystal).
- the nanocrystal surfaces are completely free of any organic contaminants or reactants which materially change their functionality. It should be further understood that incidental components that are caused to adhere to nanocrystals of the invention and do not adversely or materially affect the functioning of the inventive nanocrystals, should still be considered to be within the metes and bounds of the invention.
- the lack of added chemicals permits the growth of the metal atoms and also does not adversely affect the performance of the nanocrystals (e.g., in catalysis reactions or in biological reactions, in vivo it affects the protein corona formed around the nanoparticles/nanocrystals in, for example, serum and/or reduces toxic compounds introduced into cells or or an organism).
- protein corona formation can control location of a nanoparticle/nanocrystal in vivo, as well as control protein folding of proteins at or near the nanoparticle/nanocrystal surfaces.
- Such differences in performance may be due to such factors including, but not limited to, surface charge, surface plasmon resonance, epitaxial effects, surface double layers, zones of influence, toxic surface contaminents and others.
- Such novel shapes also affect, for example, catalysis.
- a seed crystal occurs in the process and/or a set of extended crystal planes begins to grow (e.g., homogenous nucleation) or a seed crystal is separately provided (e.g., heterogenous nucleation)
- the amount of time that a formed particle e.g., a metal atom
- the amount of time that a formed particle is permitted to dwell at or near one or more electrodes in an electrochemical process can result in the size of bi-metallic nanocrystals increasing as a function of time (e.g., metal atoms can assemble into metal nanocrystals and, if unimpeded by certain organic constituents in the liquid, they can grow into a variety of shapes and sizes).
- the amount of time that crystal nucleation/growth conditions are present can control the shape(s) and sizes(s) of grown bi-metallic nanocrystals. Accordingly, dwell time at/around electrodes, liquid flow rate(s), trough cross-sectional shape(s), etc, all contribute to nanocrystal growth conditions, as discussed elsewhere herein.
- one or more AC sources are utilized (e.g., transformer(s) 60 and power supply 501 AC).
- the rate of change from “+” polarity on one electrode to “ ⁇ ” polarity on the same electrode is known as Hertz, Hz, frequency, or cycles per second.
- the standard output frequency is 60 Hz, while in Europe it is predominantly 50 Hz.
- the frequency can also influence size and/or shape and/or presence of nanocrystals and/or ions formed according to the electrochemical techniques disclosed herein.
- Preferable frequencies are 5-1000 Hz, more typically, 20-500 Hz, even more typically, 40-200 Hz, and even more typically, 50-100 Hz.
- nucleated or growing crystals can first have attractive forces exerted on them (or on crystal growth constituents, such as ions or atoms, taking part in forming the crystal(s)) due to, for example, unlike charges attracting and then repulsive forces being exerted on such constituents (e.g., due to like charges repelling).
- the particular waveform that is used for a specific frequency also affects nanocrystal growth conditions, and thus effects nanocrystal size(s) and/or shape(s).
- the U.S. uses a standard AC frequency of 60 Hz, it also uses a standard waveform of a “sine” wave.
- changing the waveform from a sine wave to a square wave or a triangular wave also affects nanocrystal crystallization conditions and thus affects resultant nanocrystal size(s) and shape(s).
- Preferred waveforms include sine waves, square waves and triangular waves, however hybrid waveforms should be considered to be within the metes and bounds of the invention.
- the voltage applied in the novel electrochemical techniques disclosed herein can also affect nanocrystalline size(s) and shape(s).
- a preferred voltage range is 20-2000 Volts, a more preferred voltage range is 50-1000 Volts and an even more preferred voltage range is 100-300 Volts.
- the amperages used with these voltages typically are 0.1-10 Amps, a more preferred amperage range is 0.1-5 Amps and an even more preferred amperage range is 0.4-1 Amps per electrode set under the processing parameters disclosed herein.
- the “duty cycle” used for each waveform applied in the novel electrochemical techniques disclosed herein can also affect nanocrystalline size(s) and shape(s).
- the amount of time that an electrode is positively biased can result in a first set of reactions, while a different set of reactions can occur when the electrode is negatively biased.
- size(s) and/or shape(s) of grown nanocrystals can be controlled.
- the rate at which an electrode converts to + or ⁇ is also a function of waveform shape and also influences nanocrystal size(s) and/or shape(s).
- trough member 30 or 30 a ′ and/or 30 b ′
- any one of which can produce desirable results as a function of a variety of design and production considerations.
- one or more constituents produced in the portion(s) 30 a ′, or 30 b ′ could be transient (e.g., a seed crystal or nucleation point) and/or semi permanent (e.g., grown nanocrystals present in a colloid).
- a final product e.g., properties of a final product
- transient constituents formed in a first trough member 30 a ′/ 30 b ′ can also affect subsequent bi-metallic nanocrystal formation in a second trough member 30 a ′/ 30 b ′.
- the amount of time that lapses between the production of a first aqueous product in a first trough member and wherein such first product becomes a raw material in a second trough member can also influence the bi-metallic nanocrystal suspension formed.
- the temperature of liquids entering and exiting can be monitored/controlled to maximize certain desirable processing conditions and/or desirable properties of final products and/or minimize certain undesirable products.
- processing enhancers may be selectively utilized in one or more of the portions of the different trough members.
- FIG. 6 shows a schematic view of the general apparatus utilized in accordance with the teachings of some of the preferred embodiments of the present invention.
- this FIG. 6 shows a side schematic view of the trough member 30 containing a liquid 3 therein.
- On the top of the trough member 30 rests a plurality of control devices 20 a - 20 d which are, in this embodiment, removably attached thereto.
- the control devices 20 a - 20 d may of course be permanently fixed in position when practicing various embodiments of the invention.
- control devices 20 and corresponding electrode(s) 1 and/or 5 as well as the configuration(s) of such electrodes) and the positioning or location of the control devices 20 (and corresponding electrodes 1 and/or 5 ) are a function of various preferred embodiments of the invention discussed in greater detail elsewhere herein.
- a liquid transport means 40 e.g., a liquid pump, gravity or liquid pumping means for pumping the liquid 3
- a peristaltic pump 40 for pumping the liquid 3 into the trough member 30 at a first-end 31 thereof.
- the liquid transport means 40 may include any means for moving liquids 3 including, but not limited to a gravity-fed or hydrostatic means, a pumping means, a regulating or valve means, etc. However, the liquid transport means 40 should be capable of reliably and/or controllably introducing known amounts of the liquid 3 into the trough member 30 . The amount of time that the liquid 3 is contained within the trough member 30 (e.g., at or around one or more electrode(s) 1 / 5 ) also influences the products produced (e.g., the sizes(s) and/or shapes(s) of the grown nanocrystals).
- a difference in vertical height of less than one inch between an inlet portion 31 and an outlet portion 32 , spaced apart by about 6 feet (about 1.8 meters) relative to the support surface may be all that is required, so long as the viscosity of the liquid 3 is not too high (e.g., any viscosity around the viscosity of water can be controlled by gravity flow once such fluids are contained or located within the trough member 30 ).
- the need for a greater angle ⁇ could be a result of processing a liquid 3 having a viscosity higher than water; the need for the liquid 3 to transit the trough 30 at a faster rate, etc.
- liquid 3 when viscosities of the liquid 3 increase such that gravity alone is insufficient, other phenomena such as specific uses of hydrostatic head pressure or hydrostatic pressure can also be utilized to achieve desirable fluid flow.
- additional means for moving the liquid 3 along the trough member 30 could also be provided inside the trough member 30 .
- Such means for moving the fluid include mechanical means such as paddles, fans, propellers, augers, etc., acoustic means such as transducers, thermal means such as heaters and/or chillers (which may have additional processing benefits), etc., are also desirable for use with the present invention.
- FIG. 6 also shows a storage tank or storage vessel 41 at the end 32 of the trough member 30 .
- Such storage vessel 41 can be any acceptable vessel and/or pumping means made of one or more materials which, for example, do not negatively interact with the liquid 3 (or constituents contained therein) produced within the trough member 30 .
- Acceptable materials include, but are not limited to plastics such as high density polyethylene (HDPE), glass, metal(s) (such a certain grades of stainless steel), etc.
- HDPE high density polyethylene
- the tank 41 should be understood as including a means for distributing or directly bottling or packaging the fluid 3 processed in the trough member 30 .
- FIG. 8 c shows a perspective view of the control device 20 .
- FIG. 8 c shows a base portion 25 is provided, said base portion having a top portion 25 ′ and a bottom portion 25 ′′.
- the base portion 25 is made of a suitable rigid plastic material including, but not limited to, materials made from structural plastics, resins, polyurethane, polypropylene, nylon, teflon, polyvinyl, etc.
- a dividing wall 27 is provided between two electrode adjustment assemblies. The dividing wall 27 can be made of similar or different material from that material comprising the base portion 25 .
- the step motors 21 a , 21 b could be any step motor capable of slightly moving (e.g., on a 360 degree basis, slightly less than or slightly more than 1 degree) such that a circumferential movement of the step motors 21 a / 21 b results in a vertical raising or lowering of an electrode 1 or 5 communicating therewith.
- a first wheel-shaped component 23 a is the drivewheel connected to the output shaft 231 a of the drive motor 21 a such that when the drive shaft 231 a rotates, circumferential movement of the wheel 23 a is created.
- a slave wheel 24 a is caused to press against and toward the drivewheel 23 a such that frictional contact exists therebetween.
- the drivewheel 23 a and/or slavewheel 24 a may include a notch or groove on an outer portion thereof to assist in accommodating the electrodes 1 , 5 .
- the slavewheel 24 a is caused to be pressed toward the drivewheel 23 a by a spring 285 located between the portions 241 a and 261 a attached to the slave wheel 24 a .
- a coiled spring 285 can be located around the portion of the axis 262 a that extends out from the block 261 a .
- Springs should be of sufficient tension so as to result in a reasonable frictional force between the drivewheel 24 a and the slavewheel 24 a such that when the shaft 231 a rotates a determined amount, the electrode assemblies 5 a , 5 b , 1 a , 1 b , etc., will move in a vertical direction relative to the base portion 25 .
- Such rotational or circumferential movement of the drivewheel 23 a results in a direct transfer of vertical directional changes in the electrodes 1 , 5 shown herein.
- At least a portion of the drivewheel 23 a should be made from an electrically insulating material; whereas the slavewheel 24 a can be made from an electrically conductive material or an electrically insulating material, but typically, an electrically insulating material.
- the drive motors 21 a / 21 b can be any suitable drive motor which is capable of small rotations (e.g., slightly below 1°/360° or slightly above 1°/360°) such that small rotational changes in the drive shaft 231 a are translated into small vertical changes in the electrode assemblies.
- a preferred drive motor includes a drive motor manufactured by RMS Technologies model 1MC17-S04 step motor, which is a DC-powered step motor.
- This step motors 21 a / 21 b include an RS-232 connection 22 a / 22 b , respectively, which permits the step motors to be driven by a remote control apparatus such as a computer or a controller.
- the portions 271 , 272 and 273 are primarily height adjustments which adjust the height of the base portion 25 relative to the trough member 30 .
- the portions 271 , 272 and 273 can be made of same, similar or different materials from the base portion 25 .
- the portions 274 a / 274 b and 275 a / 275 b can also be made of the same, similar or different material from the base portion 25 .
- these portions should be electrically insulating in that they house various wire components associated with delivering voltage and current to the electrode assemblies 1 a / 1 b , 5 a / 5 b , etc.
- length and width can be any dimension which accommodates the size of the step motors 21 a / 21 b , and the width of the trough member 30 .
- length should be at least as long as the trough member 30 is wide, and typically slightly longer (e.g., 10-30%).
- the width needs to be wide enough to house the step motors 21 a / 21 b and not be so wide as to unnecessarily underutilize longitudinal space along the length of the trough member 30 .
- the length is about 7 inches (about 19 millimeters) and the width is about 4 inches (about 10.5 millimeters).
- the thickness of the base member 25 is any thickness sufficient which provides structural, electrical and mechanical rigidity for the base member 25 and should be of the order of about 3 ⁇ 4′′-3 ⁇ 4′′ (about 6 mm-19 mm). While these dimensions are not critical, the dimensions give an understanding of size generally of certain components of one preferred embodiment of the invention.
- the base member 25 (and the components mounted thereto), can be covered by a suitable cover (not shown) to insulate electrically, as well as creating a local protective environment for all of the components attached to the base member 25 .
- a suitable cover can be made of any suitable material which provides appropriate safety and operational flexibility. Exemplary materials include plastics similar to that used for other portions of the trough member 30 and/or the control device 20 and are typically transparent.
- This cover member can also be made of the same type of materials used to make the base portion 25 .
- the cover can include through-holes which can be aligned with excess portions of, for example, electrodes 5 , which can be connected to, for example, a spool of electrode wire (not shown in these drawings).
- the portions 242 , 242 a and 242 b provide resilient tension for the wire 5 a or 5 b to be provided therebetween. Additionally, this control device design causes there to be an electrical connection between the power sources 60 or 501 AC and the electrodes 1 / 5 .
- the servo-motor 21 a functions as discussed above, but two electrodes are driven by a single servo drive motor 21 a . Accordingly, a single drive motor 21 a can replace two drive motors in the case of the embodiment shown in FIG. 8 j .
- all electrical connections are provided on a top surface of (i.e., the surface further away from the liquid 3 ) resulting in certain design and production advantages.
- FIG. 8 c shows a refractory material component 29 a , 29 b .
- the component 29 is made of, for example, suitable refractory component, including, for example, aluminum oxide or the like.
- the refractory component 29 may have a transverse through-hole therein which provides for electrical connections to the electrode(s) 1 and/or 5 . Further a longitudinal through-hole is present along the length of the refractory component 29 such that electrode assemblies 1 / 5 can extend therethrough.
- FIG. 8 c specifically shows one electrode(s) 1 a as extending through a first refractory portion 29 a and one electrode(s) 5 a is shown as extending through a second refractory portion 29 b . Accordingly, each of the electrode assemblies expressly disclosed herein, as well as those referred to herein, can be utilized in combination with the preferred embodiments of the control device shown herein.
- a first process involves electrically activating the electrode(s) 1 and/or 5 (e.g., applying power thereto from a preferred power source 10 ), and the second general process occurrence involves determining, for example, how much power (e.g., voltage and/or current) is applied to the electrode(s) and appropriately adjusting electrode 1 / 5 height in response to such determinations (e.g., manually and/or automatically adjusting the height of the electrodes 1 / 5 ); or adjusting the electrode height or simply moving the electrode into (e.g., progressively advancing the electrode(s) 5 through the liquid 3 ) or out of contact with the liquid 3 , as a function of time.
- power e.g., voltage and/or current
- a preferred embodiment of the invention utilizes the automatic control devices 20 shown in various figures herein.
- the electrodes 1 / 5 are monitored either by the electrical circuit diagrammed in each of FIGS. 8 d - 8 h (e.g., for electrode sets 1 / 5 that make a plasma 4 or for electrode sets 5 / 5 ); or are monitored by the electrical circuit diagrammed in each of FIGS. 8 g and 8 i for electrode sets 5 / 5 , in some embodiments herein.
- the electrical circuit of FIG. 8 h is a voltage monitoring circuit. Specifically, voltage output from each of the output legs of the secondary coil 603 in the transformer 60 are monitored over the points “P-Q” and the points “P′-Q′”. Specifically, the resistor denoted by “R L ” corresponds to the internal resistance of the multi-meter measuring device (not shown).
- the output voltages measured between the points “P-Q” and “P′-Q′” typically, for several preferred embodiments shown in the Examples later herein, range between about 200 volts and about 4,500 volts. However, higher and lower voltages can work with many of the embodiments disclosed herein.
- Desirable target voltages have been determined for each electrode set 1 and/or 5 at each position along a trough member 30 a ′. Such desirable target voltages are achieved as actual applied voltages by, utilizing, for example, the circuit control shown in FIGS. 8 d , 8 e and 8 f .
- FIGS. 8 d , 8 e and 8 f refer to sets of relays controlled by a Velleman K8056 circuit assembly (having a micro-chip PIC16F630-I/P).
- Each transformer 60 is connected electrically in a manner shown in FIG. 8 h .
- Each transformer 60 and associated measuring points “P-Q” and “P′-Q′” are connected to an individual relay.
- each relay, 501 , 502 , etc. sequentially interrogates a first output voltage from a first leg of a secondary coil 603 and then a second output voltage from a second leg of the secondary coil 603 ; and such interrogation continues onto a first output voltage from a second transformer 60 b on a first leg of its secondary coil 603 , and then on to a second leg of the secondary coil 603 , and so on.
- the computer or logic control for the disclosed interrogation voltage adjustment techniques are achieved by any conventional program or controller, including, for example, in a preferred embodiment, standard visual basic programming steps utilized in a PC. Such programming steps include interrogating, reading, comparing, and sending an appropriate actuation symbol (e.g., raise or lower an electrode relative to the surface 2 of the liquid 3 ). Such techniques should be understood by an artisan of ordinary skill.
- the output voltages measured between the points “P-Q” and “P′-Q”′ typically, for several preferred embodiments shown in the Examples later herein, range between about 0.05 volts and about 5 volts. However, higher and lower voltages can work with many of the embodiments disclosed herein. Desirable target voltages have been determined for each electrode set 5 / 5 ′ at each position along a trough member 30 b ′. Such desirable target voltages are achieved as actual applied voltages by, utilizing, for example, the circuit control shown in FIGS. 8 e , 8 f , 8 g and 8 i . These FIG. 8 refer to sets of relays controlled by a Velleman K8056 circuit assembly (having a micro-chip PIC16F630-I/P).
- the servo-motor 21 is caused to rotate at a specific predetermined time in order to maintain a desirable electrode 5 profile.
- the servo-motor 21 responds by rotating a predetermined amount in a clockwise direction. Specifically the servo-motor 21 rotates a sufficient amount such that about 0.009 inches (0.229 mm) of the electrode 5 is advanced toward and into the female receiver portion o 5 (shown, for example in FIGS. 10 b and 11 a ).
- the electrode 5 is progressively advanced through the liquid 3 .
- such electrode 5 movement occurs about every 4.3 minutes. Accordingly, the rate of vertical movement of each electrode 5 into the female receiver portion o 5 is about 1 inch (about 1.9 cm) every 8 hours.
- a substantially constant electrode 5 shape or profile is maintained by its constant or progressive advance into and through the liquid 3 . Further, once the advancing end of the electrode 5 reaches the longitudinal end of the female receiver portion o 5 , the electrode 5 can be removed from the processing apparatus. Alternatively, an electrode collecting means for collecting the “used” portion of the electrode can be provided.
- each power source 501 AC is connected electrically in a manner shown in FIGS. 8 e , 8 f , 8 g and 8 i .
- Each power source 501 AC and associated measuring points “P-Q” and “P′-Q′” are connected to two individual relays. For example, the points “P-Q” correspond to relay number 501 and 501 ′ in FIG.
- each relay, 501 / 501 ′ and 502 / 502 ′, etc. sequentially interrogates the output voltage from the power source 501 AC and then a second voltage from the same power source 501 AC, and so on.
- the computer or logic control for the disclosed electrode height adjustment techniques are achieved by any conventional program or controller, including, for example, in a preferred embodiment, standard visual basic programming steps utilized in a PC. Such programming steps include reading and sending an appropriate actuation symbol to lower an electrode relative to the surface 2 of the liquid 3 . Such techniques should be understood by an artisan of ordinary skill.
- “Substantially clean”, as used herein should be understood when used to describe nanocrystal surfaces means that the nanocrystals do not have chemical constituents adhered or attached to their surfaces in such an amount that would materially alter the functioning of the nanocrystal in at least one of its significant properties of the metallic-based nanocrystals set forth in the Examples herein.
- the metallic-based nanocrystal does not have a layer, surface or film which covers a significant portion (e.g., at least 25% of the crystal, or in another embodiment at least 50% of the crystal). It also can mean that the nanocrystal surfaces are completely free of any organic contaminants which materially change their functionality over bare gold crystal surfaces.
- processing-enhancer or “processing-enhanced” or “process enhancer” means at least one material (e.g., solid, liquid and/or gas) and typically means an inorganic material, which material does not significantly bind to the formed nanocrystals, but rather facilitates nucleation/growth during an electrochemical-stimulated growth process.
- the material serves important roles in the process including providing charged ions in the electrochemical solution to permit the crystals to be grown.
- the process enhancer is critically a compound(s) which remains in solution, and/or does not form a coating (in one embodiment an organic coating), and/or does not adversely affect the formed nanocrystals or the formed suspension(s), and/or is destroyed, evaporated, or is otherwise lost during the electrochemical crystal growth process.
- trough member as used herein should be understood as meaning a large variety of fluid handling devices including, pipes, half pipes, channels or grooves existing in materials or objects, conduits, ducts, tubes, chutes, hoses and/or spouts, so long as such are compatible with the electrochemical processes disclosed herein.
- this Example utilizes certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c , and 11 a .
- All trough members 30 a ′ and 30 b ′ in the aforementioned Figures were made from 1 ⁇ 8′′ (about 3 mm) thick plexiglass, and 1 ⁇ 4′′ (about 6 mm) thick polycarbonate, respectively.
- the support structure 34 (not shown in many of the Figures but shown in FIG. 9 ) was also made from plexiglass which was about 3 ⁇ 4′′ thick (about 6-7 mm thick).
- Each trough member 30 a ′ was integral with trough member 30 b ′.
- Each trough member portion 30 b ′ had a cross-sectional shape corresponding to FIG. 5 a .
- the relevant dimensions for trough member portion 30 b ′ are reported in Table 1 as “M” (i.e., inside width of the trough at the entrance and exact portion of the trough member 30 b ′), “LT” (i.e., transverse length or flow length of the trough member 30 b ′), “S” (i.e., the height of the trough member 30 b ′), and “d′′” (i.e., depth of the liquid 3 ′′ within the trough member 30 b ′).
- the thickness of each sidewall portion of trough 30 b ′ also measured about 1 ⁇ 4′′ (about 6 mm) thick.
- the reverse osmosis membrane utilized several thin layers or sheets of film that are bonded together and rolled in a spiral configuration around a plastic tube. (This is also known as a thin film composite or TFC membrane.)
- the RO membrane also separates out suspended materials including microorganisms that may be present in the water.
- TDS total dissolved solvents
- the dam 80 was provided near the output end 32 of the trough member 30 a ′ and assisted in creating the depth “d′” (shown in FIG. 5 b as “d”) to be about 7/6′′-1 ⁇ 2′′ (about 11-13 mm) in depth.
- the height of the dam 80 measured about 3 ⁇ 4′′ (about 6 mm) and the longitudinal length measured about 1 ⁇ 2′′ (about 13 mm).
- the width was completely across the bottom dimension “R” of the trough member 30 a ′. Accordingly, the total volume of liquid 3 ′ in the trough member 30 a ′ during operation thereof was about 2.14 in 3 (about 35 ml) to about 0.89 in 3 (about 14.58 ml).
- Other acceptable flow rates should be considered to be within the metes and bounds of the invention.
- Table 1 shows that there was a single electrode set 1 a / 5 a .
- the power source for each electrode set 1 / 5 was an AC transformer 60 .
- FIG. 7 a shows a source of AC power 62 connected to a transformer 60 .
- a capacitor 61 is provided so that, for example, loss factors in the circuit can be adjusted.
- the output of the transformer 60 is connected to the electrode(s) 1 / 5 through the control device 20 .
- a preferred transformer for use with the present invention is one that uses alternating current flowing in a primary coil 601 to establish an alternating magnetic flux in a core 602 that easily conducts the flux.
- Preferred transformer(s) 60 for use in these Examples have deliberately poor output voltage regulation made possible by the use of magnetic shunts in the transformer 60 .
- These transformers 60 are known as neon sign transformers. This configuration limits current flow into the electrode(s) 1 / 5 . With a large change in output load voltage, the transformer 60 maintains output load current within a relatively narrow range.
- the transformer 60 is known as a Balanced Mid-Point Referenced Design (e.g., also formerly known as balanced midpoint grounded). This is most commonly found in mid to higher voltage rated transformers and most 60 mA transformers. This is the only type transformer acceptable in a “mid-point return wired” system.
- the “balanced” transformer 60 has one primary coil 601 with two secondary coils 603 , one on each side of the primary coil 601 (as shown generally in the schematic view in FIG. 7 bg ). This transformer 60 can in many ways perform like two transformers.
- each secondary coil 603 is attached to the core 602 and subsequently to the transformer enclosure and the other end of the each secondary coil 603 is attached to an output lead or terminal.
- an unloaded 15,000 volt transformer of this type will measure about 7,500 volts from each secondary terminal to the transformer enclosure but will measure about 15,000 volts between the two output terminals.
- AC alternating current
- the voltage and current each start at zero, rise to a crest, fall to zero, go to a negative crest and back up to zero. This completes one cycle of a typical sine wave. This happens 60 times per second in a typical US application. Thus, such a voltage or current has a characteristic “frequency” of 60 cycles per second (or 60 Hertz) power.
- Power factor relates to the position of the voltage waveform relative to the current waveform. When both waveforms pass through zero together and their crests are together, they are in phase and the power factor is 1, or 100%.
- FIG. 7 c shows two waveforms “V” (voltage) and “C” (current) that are in phase with each other and have a power factor of 1 or 100%; whereas FIG. 7 d shows two waveforms “V” (voltage) and “C” (current) that are out of phase with each other and have a power factor of about 60%; both waveforms do not pass through zero at the same time, etc.
- the waveforms are out of phase and their power factor is less than 100%.
- the normal power factor of most such transformers 60 is largely due to the effect of the magnetic shunts 604 and the secondary coil 603 , which effectively add an inductor into the output of the transformer's 60 circuit to limit current to the electrodes 1 / 5 .
- the power factor can be increased to a higher power factor by the use of capacitor(s) 61 placed across the primary coil 601 of the transformer, 60 which brings the input voltage and current waves more into phase.
- the unloaded voltage of any transformer 60 to be used in the present invention is important, as well as the internal structure thereof.
- Desirable unloaded transformers for use in the present invention include those that are around 9,000 volts, 10,000 volts, 12,000 volts and 15,000 volts. However, these particular unloaded volt transformer measurements should not be viewed as limiting the scope acceptable power sources as additional embodiments.
- a specific desirable transformer for use in these Examples is made by Franceformer, Catalog No. 9060-P-E which operates at: primarily 120 volts, 60 Hz; and secondary 9,000 volts, 60 mA.
- each transformer assembly 60 a - 60 h (and/or 60 a ′- 60 h ′; and/or 60 a ′′- 60 h ′′) can be the same transformer, or can be a combination of different transformers (as well as different polarities).
- the wires used to attach electrode 1 to the transformer 60 were, for Examples 1-3, 99.95% (3N5) gold wire, having a diameter of about 1 mm.
- the plasma 4 was created with an electrode 1 similar in shape to that shown in FIG. 3 e , and weighed about 9.2 grams. This electrode was 99.95% pure gold.
- the other electrode 5 a measured about 1 mm thick gold wire (99.95%) and having about 9 mm submerged in the liquid 3 ′.
- the output from the trough member 30 a ′ was the conditioned liquid 3 ′ and this conditioned liquid 3 ′ flowed directly into a second trough member 30 b ′.
- the second trough member 30 b ′ shown in FIGS. 10 b and 11 a had measurements as reported in Table 1.
- This trough member 30 b ′ contained about 885 ml of liquid 3 ′′.
- Table 1 reports the electrode configuration, as shown in FIGS. 8 b and 11 a , which means seven sets of electrodes 5 / 5 ′ (shown in FIG. 8 b ) were positioned as shown in FIG.
- Each of the electrode sets 5 / 5 ′ comprised 99.99% pure gold wire measuring about 1.0 mm in diameter, as reported in Table 1.
- the length of each wire electrode 5 that was in contact with the liquid 3 ′′ (reported as “WL” in Table 1) measured about 1′′ (about 25.4 mm). Other orientations fit within the metes and bounds of this disclosure.
- the AC power source (or transformer) 501 AC illustrated in FIG. 13 , was used as the power supply for examples contained herein.
- This transformer 501 AC was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of about 2 kVA.
- each separate electrode set 5 / 5 ′ e.g., Set 2 , Set 3 -Set 8 or Set 9
- power supply 501 AC was electrically connected to each electrode set, according to the wiring diagram show in FIG. 10 a .
- Table 1 refers to each of the electrode sets by “Set #” (e.g., “Set 1 ” through “Set 8 ”). Each electrode of the 1 / 5 or 5 / 5 electrode sets was set to operate at a specific voltage. The voltages listed in Table 1 are the voltages used for each electrode set. The distance “c-c” (with reference to FIG. 6 ) from the centerline of each electrode set to the adjacent electrode set is also reported. Further, the distance “x” associated with each electrode 1 utilized is also reported. For the electrode 5 , no distance “x” is reported. Other relevant parameters are also reported in Table 1. All materials for the electrodes 1 / 5 were obtained from Hi-Rel having an address of 23 Lewis Street, Fort Erie, Ontario, Canada, L2A 2P6.
- each electrode 5 / 5 ′ was first placed into contact with the liquid 3 ′′ such that it just entered the female receiver tube o 5 .
- gold metal was removed from each wire electrode 5 which caused the electrode 5 to thin (i.e., become smaller in diameter) which changed, for example, current density and/or the rate at which gold nanoparticles were formed. Accordingly, the electrodes 5 were moved toward the female receiver tubes o 5 resulting in fresh and thicker electrodes 5 entering the liquid 3 ′′ at a top surface portion thereof.
- an erosion profile or tapering effect was formed on the electrodes 5 after some amount of processing time has passed (i.e., portions of the wire near the surface of the liquid 3 ′′ were typically thicker than portions near the female receiver tubes o 5 ), and such wire electrode profile or tapering can remain essentially constant throughout a production process, if desired, resulting in essentially identical product being produced at any point in time after an initial pre-equilibrium phase during a production run allowing, for example, the process to be cGMP under current FDA guidelines and/or be ISO 9000 compliant as well.
- the electrodes 5 / 5 were actuated or moved at a rate of about 1 inch per 8 hours. Samples were collected only from the equilibrium phase.
- the pre-equilibrium phase occurs because, for example, the concentration of nanocrystals produced in the liquid 3 ′′ increases as a function of time until the concentration reaches equilibrium conditions (e.g., substantially constant nucleation and growth conditions within the apparatus), which equilibrium conditions remain substantially constant through the remainder of the processing due to the control processes disclosed herein.
- the eight electrode sets 1 / 5 and 5 / 5 were all connected to control devices 20 through 20 g which automatically adjusted the height of, for example, each electrode 1 / 5 or 5 / 5 in each electrode set.
- Two female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- Each female receiver tube o 5 was made of polycarbonate and had an inside diameter of about 1 ⁇ 8 inch (about 3.2 mm) and was fixed in place by a solvent adhesive to the bottom portion of the trough member 30 b ′.
- each tube o 5 Holes in the bottom of the trough member 30 b ′ permitted the outside diameter of each tube o 5 to be fixed therein such that one end of the tube o 5 was flush with the surface of the bottom portion of the trough 30 b ′.
- the bottom portion of the tube o 5 is sealed.
- the inside diameters of the tubes o 5 effectively prevented any significant quantities of liquid 3 ′′ from entering into the female receiver tube o 5 . However, some liquid may flow into the inside of one or more of the female receiver tubes o 5 .
- the length or vertical height of each female receiver tube o 5 used in this Example was about 6 inches (about 15.24 cm) however, shorter or longer lengths fall within the metes and bounds of this disclosure.
- female receiver tubes o 5 are shown as being subsequently straight, such tubes could be curved in a J-shaped or U-shaped manner such that their openings away from the trough member 30 b ′ could be above the top surface of the liquid 3 ,” if desired.
- the run described in this example utilize the following processing enhancer, Specifically, about 2.0 grams/gallon (i.e., about 0.528 g/liter) of sodium hydrogen carbonate (“soda”), having a chemical formula of NaHCO 3 , was added to and mixed with the water 3 .
- the soda was obtained from Alfa Aesar and the soda had a formula weight of 84.01 and a density of about 2.159 g/cm 3 .
- a sine wave AC frequency at 60 Hz was utilized to make nanocrystal suspensions or colloids and/or ion solutions in accordance with the teachings herein.
- the AC power source 501 AC utilized a Chroma 61604 programmable AC source.
- the applied voltage was about 220 volts.
- the applied current was between about 4.5 amps and about 5.5 amps.
- Table 1 summarizes key processing parameters used in conjunction with FIGS. 9 and 10 c . Also, Table 1 discloses: 1) “Produced Au PPM” (e.g., gold nanocrystal concentrations); 2) “TEM Average Diameter” which is the mode, corresponding to the crystal diameter that occurs most frequently, determined by the TEM analysis; and 3) “Hydrodynamic radius” as measured by the Zetasizer ZS-90. These physical characterizations were performed as discussed elsewhere herein.
- TEM samples were prepared by utilizing a Formvar coated grid stabilized with carbon having a mesh size of 200.
- the grids were first pretreated by a plasma treatment under vacuum.
- the grids were placed on a microscope slide lined with a rectangular piece of filter paper and then placed into a Denton Vacuum apparatus with the necessary plasma generator accessory installed.
- the vacuum was maintained at 75 mTorr and the plasma was initiated and run for about 30 seconds.
- the system was vented and the grids removed.
- the grids were stable up to 7-10 days depending upon humidity conditions, but in all instances were used within 12 hours.
- each inventive nanocrystal suspension was placed onto each grid and was allowed to air dry at room temperature for 20-30 minutes, or until the droplet evaporated. Upon complete evaporation, the grids were placed onto a holder plate until TEM analysis was performed.
- a Philips/FEI Tecnai 12 Transmission Electron Microscope was used to interrogate all prepared samples. The instrument was run at an accelerating voltage of 100 keV. After alignment of the beam, the samples were examined at various magnifications up to and including 630,000 ⁇ . Images were collected via the attached Olympus Megaview III side-mounted camera that transmitted the images directly to a PC equipped with iTEM and Tecnai User Interface software which provided for both control over the camera and the TEM instrument, respectively.
- FIG. 11 c shows a representative TEM photomicrograph corresponding to dried solution NE10214 comprised of gold nanocrystals, dried from suspension, made according to this example.
- FIG. 11 d corresponds to the measured TEM size distribution used to calculate the TEM average diameter and referenced in Table 1.
- the pH measurements were made by using an Accumet® AR20 pH/conductivity meter wherein the pH probe was placed into a 50 mL vial containing the samples of interest and allowed to stabilize. Three separate pH measurements were then taken and averaged per sample. NE10214 had a pH of about 8.94.
- UV-VIS spectroscopy Energy absorption spectra were obtained for the samples by using UV-VIS spectroscopy. This information was acquired using a Thermofisher Evolution 201 UV-VIS spectrometer equipped with a double beam Czerny-Turner monochromator system and dual silicon photodiodes. Instrumentation was provided to support measurement of low-concentration liquid samples using one of a number of fuzed-quartz sample holders or “cuvettes.” Data was acquired over the wavelength range between about 300-900 nm with the following parameters: bandwidth of 1 nm, data pitch of 0.5 nm. A xenon flash lamp was the primary energy source. The optical pathway of the spectrometer was arranged to allow the energy beam to pass through the center of each sample cuvette.
- Sample preparation was limited to filling and capping the cuvettes and then physically placing the samples into the cuvette holder, within the fully enclosed sample compartment of the spectrometer. Optical absorption of energy of each sample was determined. Data output was measured and displayed as Absorbance Units (per Beer-Lambert's Law) versus wavelength.
- DLS dynamic light scattering
- the instrument was allowed to warm up for at least 30 min prior to the experiments.
- the measurements were made using square glass cell with 1 cm pathlength, PCS8501. The following procedure was used:
- the AAS values were obtained from a Perkin Elmer AAnalyst 400 Spectrometer system. Atomic absorption spectroscopy is used to determine concentration of species, reported in “ppm” (parts per million).
- Table 1 references the AAS concentration result as “Produced Au PPM”, with a corresponding value of 6.6 ppm
- FIG. 12 a shows the apparatus used to condition the liquid 3 . Once conditioned, the liquid 3 ′ was processed in the apparatus shown in FIG. 12 c
- the amount of NaHCO 3 processing enhancer used was about 0.375 grams/gallon (i.e., about 0.10 g/L) to about 3.0 grams/gallon (i.e., about 0.79 g/L).
- the amount of KOH processing enhancer used was about 0.95 grams/gallon (i.e., about 0.25 g/L).
- the amount of KBr processing enhancer used was about 4.6 grams/gallon (i.e., about 1.22 g/L).
- the amount of Na 3 PO 4 processing enhancer used was about 3.94 grams/gallon (i.e., about 1.04 g/L).
- the amount of KH 2 PO 4 processing enhancer was about 3.24 grams/gallon (i.e., about 0.86 g/L).
- the amount of time that the water 3 with processing enhancer was exposed to the plasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown in FIG. 12 c.
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- a second and different transformer was electrically connected to the electrodes 5 a / 5 b shown in FIG. 12 c .
- This transformer was an by AC power source having a voltage range of 0-300V, a frequency range of 47-400 Hz and a maximum power rating of 1 kVA.
- the applied voltage ranged between about 58 volts and about 300 volts.
- the diameter of the platinum wire electrodes was either about 0.5 mm or 1 mm.
- the electrodes 5 a , 5 b were electrically connected to power amplifier, as shown in FIG. 12 e .
- the power supply for the amplifier is set forth in FIG. 12 f .
- the power amplifier was driven by an external function generator connected to the input pins in the amplifier.
- the amount of platinum nanoparticles produced in the suspensions varied between about 10 ppm and about 25 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- the sizes of the nanoparticles made according to this Example are fully discussed in Tables 2 and 3 herein.
- TEM sample preparation was identical to the methods described earlier although interrogation was performed on a Philips EM 420 TEM equipped with a SIS Megaview III CCD digital camera.
- the TEM micrographs show that the particles have an average diameter of less than 10 nm.
- FIG. 14 shows a representative TEM Photomicrograph of platinum nanocrystals, dried from suspension GRPt-621, made according to this example.
- FIG. 12 a shows the apparatus used to condition the liquid 3 . Once conditioned, the liquid 3 ′ was processed in the apparatus shown in FIG. 12 d.
- the amount of KBr processing enhancer used was about 4.6 grams/gallon (i.e., about 1.2 grams/Liter) or about 1.4 g/gal (i.e., about 0.4 g/L).
- the amount of Na 3 PO 4 processing enhancer used was about 1.9 grams/gallon (i.e., about 0.5 g/L).
- the amount of time that the water 3 with each processing enhancer was exposed to the plasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown in FIG. 12 d.
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- a power supply (shown in FIG. 12 f ) was utilized to apply a sinusoidal voltage with a frequency of about 2.5 Hz to the electrodes 5 a and 5 b .
- the electrodes were electrically connected to a power amplifier, as shown in FIG. 12 e .
- the distance between the electrodes was fixed in all suspensions at approximately 7 mm.
- the amplifier was driven by an external function generator connected to the input pins in the amplifier.
- Suspensions PRX37-01 and PRX37-02 show that for a given conductivity of water 3 , and a given voltage applied at a fixed distance to electrodes 5 a and 5 b , the amount of platinum in the final suspension increased as the amount of KBr processing enhancer was increased.
- the average hydrodynamic radii of the formed particles in water were analyzed with the dynamic light scattering technique discussed elsewhere herein.
- the hydrodynamic radius is not reported (NR) for formulation PRX37-02 because the transmission amount reported in the DLS device was 100%, indicating a high presence of dissolved platinum species (e.g., ions).
- FIG. 15 a shows a representative TEM Photomicrograph of platinum nanocrystals, dried from suspension PRX37-03, made according to this Example 3. Table 4 is included to show the relevant processing conditions used as well as certain resultant physical properties of the formulation PRX37.
- this Example utilizes certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 d and 11 b .
- the AC power source (or transformer) 501 AC illustrated in FIG. 13 , was used as the power supply for the examples contained herein, while the function generator 501 FG was sometimes used (as disclosed herein) to drive the AC power source 501 AC.
- This transformer 501 AC was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of about 2 kVA. The precise electrical connections are discussed elsewhere herein. Control devices 20 , as illustrated in FIGS.
- the amount of NaHCO 3 (Fisher Scientific, Cat #S631-3) processing enhancer used was about 2.5 grams/gallon (i.e., about 0.67 g/L) to about 3.5 grams/gallon (i.e., about 0.93 g/L).
- the amount of KHCO 3 processing enhancer used was about 2.31 grams/gallon (i.e., about 0.61 g/L).
- the amount of NaOH processing enhancer used was about 0.70 grams/gallon (i.e., about 0.19 g/L).
- the amount of KOH processing enhancer used was about 0.72 grams/gallon (i.e., about 0.19 g/L).
- the amount of NaBr processing enhancer was about 2.18 grams/gallon (i.e., about 0.58 g/L).
- the amount of KBr processing enhancer was about 2.04 grams/gallon (i.e., about 0.54 g/L).
- the amount of Na 2 PO 4 processing enhancer was about 1.08 grams/gallon (i.e., about 0.29 g/L).
- the amount of NaCl processing enhancer was about 1.27 grams/gallon (i.e., about 0.34 g/L).
- the amount of CaCl 2 processing enhancer was about 1.16 grams/gallon (i.e., about 0.31 g/L).
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- the AC power source 501 AC utilized a Chroma 61604 programmable unit.
- sine wave AC frequencies at 5 Hz and 80 Hz were utilized to make nanocrystal suspensions or colloids and/or ions, in accordance with the teachings herein.
- the applied voltage was about 175 volts.
- the function generator 501 FG provided sine waves at frequencies less than 15 Hz to the AC power source 501 AC, which subsequently amplified the input signal to about 175 volts at different frequencies.
- the applied current varied between about 3.0 amps and about 6.5 amps.
- FIG. 16 shows a representative TEM Photomicrograph of platinum nanocrystals, dried from suspension PB-13, made according to this Example 4.
- the amount of platinum nanoparticles or ions produced in the formulations varied between about 1.0 ppm and about 15 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Tables 5-8 summarize key processing parameters used in conjunction with FIGS. 9 a and 10 d . Also, Tables 5-8 disclose: 1) resultant “ppm” (e.g., platinum nanocrystal/ion concentrations.)
- Figure 8b 8b 8b 8b 8b 8b 8b 8b 8b 8b 8b Produced Pt PPM 8.1 11.8 2.3 5.9 2.4 7.0 Output Temp ° C. at 32 70 70 65 63 66 64 Dimensions Plasma 4 Figs. 9 9 9 9 9 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm) 36/914 36/914 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/2
- Figure 8b 8b 8b 8b 8b 8b 8b 8b 8b 8b 8b Produced Pt PPM 2.4 7.0 1.1 3.6 1.4 3.9 Output Temp ° C. at 32 68 66 60 60 63 60 Dimensions Plasma 4 Figs. 9 9 9 9 9 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm) 36/914 36/914 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25
- Figure 8b 8b 8b 8b 8b 8b 8b 8b 8b 8b 8b Produced Pt PPM 2.5 9.9 2.2 7.1 1.6 4.1 Output Temp ° C. at 32 68 70.5 61.5 64 61 61 Dimensions Plasma 4 Figs. 9 9 9 9 9 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm) 36/914 36/914 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25
- this Example utilizes certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 d and 11 b .
- the AC power source (or transformer) 501 AC illustrated in FIG. 13 , was used as the power supply for the examples contained herein, while the function generator 501 FG was sometimes used (as disclosed herein) to drive the AC power source 501 AC.
- This transformer 501 AC was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of about 2 kVA. The precise electrical connections are discussed elsewhere herein. Control devices 20 , illustrated in FIGS.
- sine wave AC frequencies as low as about 1 Hz and as high as about 200 Hz were utilized to make nanocrystal suspensions or colloids and/or ions, in accordance with the teachings herein.
- the AC power source 501 AC utilized a Chroma 61604 programmable AC source.
- the applied voltage was about 175 volts with a corresponding sine wave at six different frequencies of about 15, 40, 60, 80, 100 and 200 Hz.
- the function generator 501 FG provided sine waves at frequencies less than 15 Hz to the power supply 501 AC which subsequently amplified the input signal to about 175V at different frequencies, namely 1 Hz and 5 Hz.
- the applied current varied between about 4.5 amps and 6.0 amps.
- the amount of platinum nanoparticles and/or ions produced in the formulations varied between about 7.0 ppm and about 15 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Tables 9-10 summarize key processing parameters used in conjunction with FIGS. 9 and 10 d . Also, Tables 9-10 disclose: 1) resultant “ppm” (i.e., platinum concentrations.)
- FIG. 17 contains the UV-Vis data collected for the samples above, specifically displaying the 265 nm-750 nm range.
- Figure 8b 8b 8b 8b 8b 8b 8b 8b 8b 8b 8b Produced Pt PPM 9.7 8.6 8.7 12.1 14.6 7.7 Output Temp ° C. at 32 72 72 72 71 72 71 Dimensions Plasma 4 Figs. 9 9 9 9 9 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm) 36/914 36/914 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/25 1/2
- FIG. 12 a shows the apparatus used to condition the liquid 3 . Once conditioned, the liquid 3 ′ was processed in the apparatus shown in FIG. 12 c or 12 d , for platinum ions/particles and bi-metallic nanocrystals, respectively. The overall process of creating a bi-metallic nanocrystal suspension is described below and is summarized in Table 11.
- platinum ions and/or particles were created in water by the following process. Approximately 4.0 grams/gallon (i.e., about 1.06 mg/mL) of processing enhancer baking soda (i.e., NaHCO 3 ) was added to about 1 gallon of de-ionized water. The amount of time that the water 3 with processing enhancer was exposed to the plasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown in FIG. 12 c.
- processing enhancer baking soda i.e., NaHCO 3
- the applied voltage for each plasma 4 created at electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. Note that in Table 11 (and elsewhere herein) the reference to “GZA” is synonomous with creation of plasma 4 .
- a second and different transformer was electrically connected to the electrodes 5 a / 5 b shown in FIG. 12 c .
- This transformer was a hy AC power source having a voltage range of about 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA.
- the applied voltage was about 100 volts with a frequency of about 60 hertz for approximately a 2-hour operating time.
- the diameter of the platinum wire electrodes was 1 mm.
- the length of the platinum wires was about 51 mm.
- the platinum species and water formulation (raw material) prepared above was mixed with an equal amount of conditioned water, which conditioned water 3 ′ was achieved with a platinum electrode 1 creating a plasma 4 for about 30 minutes, and processing enhancer NaHCO 3 0.5 g/gallon (0.132 mg/mL) NaHCO 3 ) at a ratio of 1:1 to a total volume of about 800 mL.
- the liquid 3 ′ was then processed via the apparatus in FIG. 12 d with gold electrodes (99.99%, about 0.5 mm diameter and a length of about 6.25 in (15.88 cm) for about 40 minutes, with a hy AC power source having an applied voltage of about 160 volts and about 47 hertz.
- the hydrodynamic radius of the bi-metallic nanocrystals made was about 14.7 nm as measured by ViscoTek.
- the suspension contained about 16.1 ppm of Au and about 2.1 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- FIG. 18 shows a representative TEM Photomicrograph of the bi-metallic nanocrystal suspension dried from formulation 110910-4, which was made by techniques equivalent to those discussed elsewhere herein.
- FIG. 12 g contains the UV-Vis data collected for this sample (111710-a), specifically displaying the 350-900 nm range.
- DLS dynamic light scattering
- the instrument was allowed to warm up for at least 30 min prior to the experiments.
- the measurements were made using 12 ⁇ l quartz cell. The following procedure was used:
- the dynamic light scattering particle size information is different from the TEM measured histograms because dynamic light scattering uses algorithms that assume the nanocrystals are all spheres (which they are not) as well as measures the hydrodynamic radius (e.g., the nanocrystal's influence on the water is also detected and reported in addition to the actual physical radii of the particles). Accordingly, it is not surprising that there is a difference in the reported particle sizes between those reported in the TEM histogram data and those reported in the dynamic light scattering data, just as in the other Examples included herein.
- FIG. 12 a shows the apparatus used to condition the liquid 3 . Once conditioned, the liquid 3 ′ was processed in the apparatus shown in FIG. 12 c or 12 d , for platinum ions/particles and bi-metallic nanocrystals, respectively. The overall process of creating a bi-metallic nanocrystal suspension is described below and is summarized in Table 12.
- platinum ions and/or particles were created in water by the following process. Approximately 4.0 grams/gallon (i.e., about 1.06 mg/mL) of processing enhancer baking soda (i.e., NaHCO 3 ) was added to about 1 gallon of de-ionized water. The amount of time that the water 3 with processing enhancer was exposed to the plasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown in FIG. 12 c . Note that in Table 12 (and elsewhere herein) the reference to “GZA” is synonomous with creation of plasma 4 .
- processing enhancer baking soda i.e., NaHCO 3
- the applied voltage for each plasma 4 created at electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- a second and different transformer was electrically connected to the electrodes 5 a / 5 b shown in FIG. 12 c .
- This transformer was a hy AC power source having a voltage range of 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA.
- the applied voltage was about 100 volts with a frequency of about 60 hertz for approximately a 2-hour operating time.
- the diameter of the platinum wire electrodes was about 1 mm.
- the platinum species and water formulation (raw material) prepared above was mixed with about 6.29 mM NaHCO 3 at a ratio of about 3:1 to create a total volume of about 3785 mL.
- This liquid 3 ′ was then processed via the apparatus shown in FIG. 12 d with gold electrodes (99.99%, 0.5 mm) for about 90 minutes, with a hy AC power source having an applied voltage of about 200 volts and about 60 hertz.
- the hydrodynamic radius of the bi-metallic nanocrystals made was about 15.4 nm as measured by ViscoTek.
- the suspension contained about 5.6 ppm of Au and about 1.6 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- FIG. 19 shows a representative TEM Photomicrograph of the bi-metallic nanocrystal suspension dried from formulation 101910-6, which was obtained by techniques equivalent to those disclosed elsewhere herein.
- FIG. 12 a shows the apparatus used to condition the liquid 3 . Once conditioned, the liquid 3 ′ was processed in the apparatus shown in FIG. 12 c or 12 d , for platinum ions/particles and bi-metallic nanocrystals, respectively. The overall process of creating a bi-metallic nanocrystal suspension is described below and is summarized in Table 13.
- platinum ions and/or particles were created in water by the following process. Approximately 0.580 grams/gallon (i.e., about 0.153 mg/mL) of processing enhancer potassium hydroxide (i.e., KOH) was added to about 1 gallon of de-ionized water. The amount of time that the water 3 with processing enhancer was exposed to the plasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown in FIG. 12 c.
- processing enhancer potassium hydroxide i.e., KOH
- the applied voltage for each plasma 4 created at electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. Note that in Table 13 (and elsewhere herein) the reference to “GZA” is synonomous with creation of plasma 4 .
- a second and different transformer was electrically connected to the electrodes 5 a / 5 b shown in FIG. 12 c .
- This transformer was a hy AC power source having a voltage range of about 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA.
- the applied voltage was about 260 volts with a frequency of about 60 hertz for approximately a 2-hour operating time.
- the diameter of the platinum wire electrodes was about 1 mm.
- the length of the platinum wires was about 51 mm (2.01 inch/5.1 cm).
- the platinum species and water formulation (raw material) prepared above was further processed as described below.
- the liquid 3 ′ was then processed via the apparatus in FIG. 12 d with gold electrodes (99.99%, about 0.5 mm diameter and about 6.25 inches (15.88 cm) total length for about 2 hours, with a hy AC power source having an applied voltage of about 180 volts and about 47 hertz.
- the hydrodynamic radius of the gold/platinum material made was about 12.5 nm as measured by ViscoTek.
- the suspension contained about 8.0 ppm of Au and about 1.8 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- FIG. 20 shows a representative TEM Photomicrograph of the bi-metallic nanocrystal suspension dried from formulation ID #122310A, made according to this Example 8.
- FIG. 12 a shows the apparatus used to condition the liquid 3 . Once conditioned, the liquid 3 ′ was processed in the apparatus shown in FIG. 12 c or 12 d , for platinum ions/nanocrystal and for gold nanocrystals, respectively.
- the overall process of creating the individual nanocrystal suspensions and thus mixing them together to form a bi-metallic nanoparticle suspension is described below and is summarized in Table 14.
- platinum ions and/or particles were created in water by the following process. Approximately 4.0 grams/gallon (i.e., about 1.06 mg/mL) of processing enhancer baking soda (i.e., NaHCO 3 ) was added to about 1 gallon of de-ionized water. The amount of time that the water 3 with processing enhancer was exposed to the plasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown in FIG. 12 c.
- processing enhancer baking soda i.e., NaHCO 3
- the applied voltage for each plasma 4 created at electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- a second and different transformer was electrically connected to the electrodes 5 a / 5 b shown in FIG. 12 c .
- This transformer was a hy AC power source having a voltage range of about 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA.
- the applied voltage was about 130 volts with a frequency of about 60 hertz for approximately a 30-minute operating time.
- the diameter of the platinum wire electrodes was about 1 mm.
- the length of the platinum wires was about 51 mm.
- the platinum species and water material was set aside.
- a separate suspension of gold nanocrystals was prepared as follows. Approximately 1.0 gram/gallon (i.e., about 0.264 mg/mL) of processing enhancer baking soda (i.e., NaHCO 3 ) was added to about 1 gallon of de-ionized water. The amount of time that the water 3 with processing enhancer was exposed to the plasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown in FIG. 12 c.
- processing enhancer baking soda i.e., NaHCO 3
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- a second and different transformer was electrically connected to the electrodes 5 a / 5 b shown in FIG. 12 d .
- This transformer was a hy AC power source having a voltage range of about 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA.
- the applied voltage was about 300 volts with a frequency of about 60 hertz for approximately a 30-minute operating time.
- the diameter of the gold wire electrodes was about 0.5 mm.
- the length of the gold wire was about 159 mm.
- the separately prepared Pt and Au water-based materials Pt formulation and Au formulation prepared above were mixed together in the presence of a hydrogen peroxide catalyst (H 2 O 2 , Alfa Aesar Cat #L14000) and then studied. Specifically, about 300 mL of Pt formulation 062810 and about 700 mL of Au formulation 061610 were combined and approximately 250 ⁇ L of H 2 O 2 0.8 v/v % was added. The measured hydrodynamic radius of the combined formulations was about 35 nm as measured by ViscoTek. The resulting suspension contained about 8.0 ppm of Au and about 1.8 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- H 2 O 2 Alfa Aesar Cat #L14000
- sample 111710-9 made substantially identically to sample 112210-1 as described in Example 6, had identifiable platinum present on the formed bi-metallic nanocrystals.
- the measured hydrodynamic radius of the bi-metallic nanocrystals was about 14.7 nm as measured by ViscoTek.
- the suspension contained about 16.1 ppm of Au and about 2.1 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Representative FIGS. 21 a - 21 b illustrate the structures formed when prepared as described above. It is evident through energy dispersive analysis that platinum is present at detectable concentrations, as indicated by representative FIGS. 22 a - 22 b.
- TEM samples were prepared by utilizing a lacey Formvar/carbon-coated copper grid having a mesh size of 200. Approximately 1-3 ⁇ L of each inventive nanocrystal suspension, colloid and/or solution was placed onto each grid and was allowed to air dry at room temperature for about 20-30 minutes, or until the droplet evaporated. Upon complete evaporation, the grids were placed onto a holder plate until TEM analysis was performed.
- a Philips CM300 FEG High Resolution Transmission Electron Microscope equipped with an Oxford thin window light element detector and Emispec ES vision 4 processor, was used to interrogate all prepared samples. The instrument was run at an accelerating voltage of about 297 kV. After alignment of the electron beam, the prepared samples were examined at various magnifications up to and including 800,000 ⁇ . Images were collected via the integrated CCD camera mounted at the back of the Gatan Image Filter (GIF) which is linked directly to a PC equipped with Digital Micrograph Software and Emispec ES Vision 4.0 software.
- GIF Gatan Image Filter
- this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 d and 11 b .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. The precise electrical connections are described elsewhere herein.
- Control devices 20 as illustrated in FIGS. 8 c and 8 j were connected to the electrodes 1 / 5 and 5 / 5 , respectively.
- the amount of potassium hydroxide (Fisher Scientific, Cat #P250-500) processing enhancer used in Run ID “PB-53” was about 0.604 grams/gallon (i.e., about 0.16 mg/mL.).
- the feed electrodes were platinum wires (1 mm/0.040′′ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- the AC power source 501 AC utilized a Chroma 61604 programmable unit.
- sine wave AC frequencies at 80 Hz were utilized to make suspensions of Pt ions and/or Pt colloids, in accordance with the teachings herein.
- the applied voltage was 215 volts with an applied current between about 4.0 amps and about 5.0 amps.
- the resulting Pt-water-based material was then allowed to cool to approximately 50 degrees Celsius. At that point the Pt-water-based material was fed into another separate and different trough unit as described below.
- this additional trough which utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a .
- Electrical device 501 AC illustrated in FIG. 13 was used as the power supply for examples contained herein, while function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found elsewhere herein.
- Control devices 20 illustrated in FIGS.
- the eight electrode sets 1 / 5 and 5 / 5 were all connected to control devices 20 and 20 i which automatically adjusted the height of, for example, each electrode 5 / 5 in each electrode set 5 / 5 ; had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- a sine wave AC frequency at 60 Hz was utilized to form the bi-metallic nanocrystalline suspension in accordance with the teachings herein.
- the platinum-water based material “PB-53,” as discussed above, was fed as a raw material via pump 40 into plasma trough section 30 a ′ as illustrated in FIG. 10 c .
- the AC power source 501 AC utilized a Chroma 61604 programmable AC source.
- the applied voltage was about 260 volts for approximately two minutes followed by about 220 volts for the duration of the run.
- the applied current varied between about 4 amps and about 5 amps.
- TEM Transmission electron microscopy
- the total amount of platinum species and gold species contained within this bi-metallic nanocrystalline suspension was about 1.6 ppm and 7.7 ppm, respectively, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 15 summarizes key processing parameters used in conjunction with FIGS. 9 and 10 b . Table 15 also discloses: 1) resultant “ppm” (i.e., atomic platinum and gold concentrations.)
- FIG. 12 a shows the apparatus used to condition the liquid 3 . Once conditioned, the liquid 3 ′ was processed in the apparatus shown in FIG. 12 c or 12 d , for platinum ions/particles and bi-metallic nanocrystals, respectively. The overall process created a bi-metallic nanocrystal suspension, as described below and summarized in Table 16.
- platinum ions and/or particles were prepared by the following process. Approximately 0.580 grams/gallon (i.e., about 0.153 mg/mL) of processing enhancer potassium hydroxide (i.e., KOH) was added to 1 gallon of de-ionized water. The amount of time that the water 3 with processing enhancer was exposed to the plasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown in FIG. 24 c.
- processing enhancer potassium hydroxide i.e., KOH
- the applied voltage for the plasma 4 made by the electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. Note that in Table 16 (and elsewhere herein) the reference to “GZA” is synonomous with creation of plasma 4 .
- a second and different transformer was electrically connected to the electrodes 5 a / 5 b shown in FIG. 12 c .
- This transformer was an hy AC power source having a voltage range of 0-300V, a frequency range of 47-400 Hz and a maximum power rating of 1 kVA.
- the applied voltage was about 100 volts with a frequency of 60 hertz for about 3 hours of operation.
- the diameter of the platinum wire electrodes was about 1 mm.
- the platinum species and water material prepared above was further processed as described below.
- the platinum species and water material was then processed via the apparatus in FIG. 12 d with gold electrodes (99.99%, 0.5 mm) for about 3 hours, with an hy AC power source having an applied voltage of about 180 volts and about 47 hertz.
- the average radius of the bi-metallic nanocrystals produced was about 14.6 nm as measured by ViscoTek.
- the suspension contained about 7.3 ppm of Au and about 1.2 ppm of Pt, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- FIGS. 26 a and 26 b show representative TEM Photomicrographs and energy-dispersive x-ray spectra of the formed bi-metallic nanocrystals, respectively, dried from suspension ID #PGB002, made according to this Example 11.
- this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 d and 11 b .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments.
- Control devices 20 illustrated in FIGS. 8 c and 8 j , were connected to the electrodes 1 / 5 and 5 / 5 , respectively.
- sine wave AC frequencies at 5 Hz were utilized to make Pt species in water in accordance with the teachings herein.
- the function generator 501 FG provided sine waves at frequencies less than 15 Hz to power supply 501 AC, Chroma 61604 programmable AC source, which subsequently amplified the input signal to about 150V.
- the applied current varied between about 5.0 amps to about 6.5 amps.
- the amount of platinum species produced in the water was about 15.9 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 17 summarizes key processing parameters used in conjunction with FIGS. 9 and 10 d . Table 17 also discloses resultant “ppm” (i.e., atomic platinum nanocrystal concentrations.)
- this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 d and 11 b .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments.
- Control devices 20 illustrated in FIGS. 8 c and 8 j were connected to the electrodes 1 / 5 and 5 / 5 , respectively.
- sine wave AC frequencies at 5 Hz were utilized to make Pt species in water in accordance with the teachings herein.
- the function generator 501 FG provided sine waves at frequencies less than 15 Hz to power supply 501 AC, Chroma 61604 programmable AC source, which subsequently amplified the input signal to about 175V.
- the applied current varied between about 4.0 amps to about 6.5 amps.
- the amount of platinum species produced in the water suspensions was about 7.8 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 18 summarizes key processing parameters used in conjunction with FIGS. 9 and 10 d . Table 18 also discloses resultant “ppm” (i.e., atomic platinum nanocrystal concentrations.)
- this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.
- Control devices 20 illustrated in FIGS.
- Electrodes 8 c and 8 j were connected to the electrodes 1 / 5 and 5 / 5 , respectively, and electrodes 5 / 5 were actuated at a rate of about 1′′ per 8 hours.
- the eight electrode sets 1 / 5 and 5 / 5 were all connected to control devices 20 and 20 i which automatically adjusted the height of, for example, each electrode or 5 / 5 in each electrode set 5 / 5 ; had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- the amount of potassium hydroxide (Fisher Scientific, Cat #P250-500) processing enhancer used in Run ID “PB-106-2” was about 0.450 grams/gallon (i.e., about 0.119 mg/mL).
- the amount of sodium bicarbonate (Fisher Scientific, Cat #S631-3) used in Run ID “PB-106-2” was about 0.850 grams/gallon (i.e., about 0.22 mg/mL).
- the feed electrodes were platinum wires (1 mm/0.040′′ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- the AC power source 501 AC utilized a Chroma 61604 programmable unit.
- sine wave AC frequencies at 80 Hz were utilized to make at least one platinum species in water in accordance with the teachings herein.
- the applied voltage was about 215 volts with an applied current between about 4.0 amps and about 7.0 amps.
- this second trough utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.
- Control devices 20 illustrated in FIGS.
- each electrode 5 / 5 in each electrode set 5 / 5 had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- a sine wave AC frequency at 60 Hz was utilized to make a gold nanocrystal suspension or colloid or ion, in accordance with the teachings herein.
- the platinum-water based material “PB-106-2,” as discussed above, was fed via pump 40 into plasma trough section 30 a ′ as illustrated in FIG. 10 c .
- the AC power source 501 AC utilized a Chroma 61604 programmable AC source.
- the applied voltage was about 260 volts for approximately two minutes followed by about 220 volts for the duration of the run.
- the applied current varied between about 4 amps and about 7 amps.
- the total amount of platinum and gold contained within the bi-metallic nanocrystal suspension this material was about 3.0 ppm and 9.2 ppm, respectively, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 19 summarizes key processing parameters used in conjunction with FIGS. 9 and 10 b . Table 19 also discloses: 1) resultant “ppm” (i.e., atomic platinum and gold concentrations.)
- Figure 8b 8b Produced Au/Pt PPM NA/3.0 9.2/3.0 Hydrodynamic Radius (nm) N/A 15.39 Zeta Potential (mV) N/A ⁇ 53.0 Dimensions Plasma 4 Figs. 9 9 Process Figures 10c, 11a 10c, 11a M (in/mm) 1.5/38 1.5/38 LT (in/mm) 36/914 36/914 d (in/mm) 1/25 1/25 S (in/mm) 1.5/38 1.5/38 Total Curr.
- zeta-Sizer “Nano-ZS” produced by Malvern Instruments was utilized to determine zeta potential (the specifics of which are described earlier herein). For each measurement a 1 ml sample was filled into clear disposable zeta cell DTS1060C. Dispersion Technology Software, version 5.10 was used to run the Zeta-Sizer and to calculate the zeta potential. The following settings were used: dispersant—water, temperature—25° C., viscosity—0.8872 cP, refraction index—1.330, dielectric constant—78.5, approximation model—Smoluchowski. Three replications of 60 runs per individual replicate were performed for each sample.
- FIG. 27 contains the UV-Vis data collected for this sample (GPB-032), specifically displaying the 350-900 nm range.
- this example utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.
- Control devices 20 illustrated in FIGS.
- each electrode 5 / 5 in each electrode set 5 / 5 had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- the amount of sodium bicarbonate (Fisher Scientific, Cat #S631-3) used in Run ID “PB-74” was about 2.5 grams/gallon (i.e., about 0.66 g/L).
- the feed electrodes were platinum wires (1 mm/0.040′′ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- the AC power source 501 AC utilized a Chroma 61604 programmable unit.
- sine wave AC frequencies at 80 Hz were utilized to make at least one platinum species in water, in accordance with the teachings herein.
- the applied voltage was 175 volts with an applied current between about 4.0 amps and about 7.0 amps.
- this second trough utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.
- Control devices 20 illustrated in FIGS.
- each electrode 5 / 5 in each electrode set 5 / 5 had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- a sine wave AC frequency at 60 Hz was utilized to make a gold nanocrystal suspension or colloid or ion, in accordance with the teachings herein.
- the platinum-water based material “PB-74,” as discussed above, was fed via pump 40 into plasma trough section 30 a ′ as illustrated in FIG. 10 b .
- the AC power source 501 AC utilized a Chroma 61604 programmable AC source.
- the applied voltage was initially set to 200 volts but was set to 165 volts due to the initial current reading falling out of the normal range, typically between 2.5 A-3.5 A.
- the applied current varied between about 4 amps and about 7 amps.
- the total amount of atomic platinum and gold contained within the bi-metallic nanocrystal suspension was about 1.7 ppm and 7.8 ppm, respectively, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. It should be noted that this particular Au—Pt bi-metallic nanocrystal suspension was not stable as it settled over a period of time no later than four months after production. Accordingly, under certain sets of processing conditions, sodim bicarbonate by itself, without the addition of KOH or other suitable processing enhancers does not promote the development of highly stable Au—Pt bi-metallic nanocrystal suspensions. However, these suspensions could be suitable for some purposes.
- Table 20 summarizes key processing parameters used in conjunction with FIGS. 9 and 10 b . Table 20 also discloses: 1) resultant “ppm” (i.e., atomic platinum and gold concentrations.) and 2) “Hydrodynamic Radius” (nm).
- this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.
- Control devices 20 illustrated in FIGS.
- each electrode 5 / 5 in each electrode set 5 / 5 had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- the amount of potassium hydroxide (Fisher Scientific, Cat #P250-500) processing enhancer used in Run IDs “PB-83, 85, 87, and 88” was about 0.450 grams/gallon (i.e., about 0.12 mg/mL.).
- the amount of sodium bicarbonate (Fisher Scientific, Cat #S631-3) used in Run IDs “PB-83, 85, 87, and 88” was about 0.850 grams/gallon (i.e., about 0.22 mg/mL).
- the feed electrodes were platinum wires (1 mm/0.040′′ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- the AC power source 501 AC utilized a Chroma 61604 programmable unit.
- sine wave AC frequencies at 80 Hz were utilized to at least one platinum species in water in accordance with the teachings herein.
- the applied voltage was about 215 volts with an applied current between about 4.0 amps and about 7.0 amps.
- this second trough utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.
- Control devices 20 illustrated in FIGS.
- each electrode 5 / 5 in each electrode set 5 / 5 had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- a sine wave AC frequency at 5 Hz-200 Hz was utilized to make gold nanocrystal suspensions or colloids or ions, in accordance with the teachings herein.
- the platinum-water based material “PB-83, 85, 87, and 88,” as discussed above, was fed via pump 40 into plasma trough section 30 a ′ as illustrated in FIG. 10 b .
- the AC power source 501 AC utilized a Chroma 61604 programmable AC source.
- the applied voltage was about 260 volts for approximately two minutes followed by about 220 volts for the duration of the run.
- the applied current varied between about 4 amps and about 7 amps.
- Tables 21a, 21b and 21c The total amount of atomic platinum and gold contained within the bi-metallic nanocrystal suspension are outlined in Tables 21a, 21b and 21c.
- Table 21a outlines the platinum run conditions used to form the platinum species in water and
- Tables 21b and 21c outline the run conditions used to form the Au—Pt bi-metallic nanocrystal suspensions.
- Table 21a summarizes key processing parameters used in conjunction with FIGS. 9 and 10 c .
- Tables 21a, 21b and 21c also disclose: 1) Resultant “ppm” (i.e., atomic platinum and gold concentrations), 2) Hydrodynamic radius, and 3) Zeta Potential.
- FIG. 28 a contains the UV-Vis data collected for thes samples (PGT024, PGT025, PGT026), specifically displaying the 350-900 nm range.
- FIG. 28 b contains the UV-Vis data collected for these samples (GPB-017, GPB-018, GPB-019, GPB-020, GPB-023), specifically displaying the 350-900 nm range.
- a variety of Au—Pt bi-metallic nanocrystal suspensions were prepared at frequencies, as described in this Example, between the range of about 5 Hz-200 Hz.
- a representative comparison of particle size versus frequency is illustrated in FIG. 28 c .
- this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a to make Au—Pt bi-metallic nanocrystal suspensions.
- Electrical device 501 AC illustrated in FIG. 13 , was used as the power supply for examples contained herein, while function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.
- Control devices 20 illustrated in FIGS.
- each electrode 5 / 5 in each electrode set 5 / 5 had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- the amount of potassium hydroxide (Fisher Scientific, Cat #P250-500) processing enhancer used in Run ID “PB-118” was about 0.450 grams/gallon (i.e., about 0.12 mg/mL.).
- the amount of sodium bicarbonate (Fisher Scientific, Cat #S631-3) used in Run ID “PB-118” was about 0.850 grams/gallon (i.e., about 0.22 mg/mL).
- the feed electrodes were platinum wires (1 mm/0.040′′ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- the applied voltage for each plasma 4 made by electrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein.
- the AC power source 501 AC utilized a Chroma 61604 programmable unit.
- sine wave AC frequencies at 80 Hz were utilized to make at least one platinum species in water, in accordance with the teachings herein.
- the applied voltage was about 215 volts with an applied current between about 4.0 amps and about 7.0 amps.
- this second trough utilized certain embodiments of the invention associated with the apparatuses generally shown in FIGS. 9 , 10 c and 11 a .
- Electrical device 501 AC illustrated in FIG. 13
- function generator 501 FG was sometimes used to drive 501 AC.
- This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.
- Control devices 20 illustrated in FIGS.
- each electrode 5 / 5 in each electrode set 5 / 5 had 2 female receiver tubes o 5 a /o 5 a ′-o 5 g /o 5 g ′ which were connected to a bottom portion of the trough member 30 b ′ such that the electrodes in each electrode set 5 / 5 could be removably inserted into each female receiver tube o 5 when, and if, desired.
- a sine wave AC frequency at 60 Hz was utilized to make a gold nanocrystal suspension or colloid or ion, in accordance with the teachings herein.
- the platinum-water based material “PB-118,” as discussed above, was fed via pump 40 into plasma trough section 30 a ′ as illustrated in FIG. 10 c .
- the AC power source 501 AC utilized a Chroma 61604 programmable AC source.
- the applied voltage was about 260 volts for approximately two minutes followed by about 220 volts for the duration of the run.
- the applied current varied between about 4 amps and about 7 amps.
- the total amount of atomic platinum and gold contained within the bi-metallic nanocrystalline suspension was about 3.2 ppm and 9.3 ppm, respectively, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 23 summarizes key processing parameters used in conjunction with FIGS. 9 and 11 a . Table 23 also discloses: 1) resultant “ppm” (i.e., atomic platinum and gold concentrations.), 2) “Hydrodynamic Radius” and 3 ) “Zeta Potential.”
- FIGS. 29 a and 29 c are representative TEM micrographs.
- FIGS. 29 b and 29 d are representative EDS spectra of dried nanocrystals in FIGS. 29 a and 29 c .
- FIGS. 29 e , 29 f and 29 g are STEM mappings of dried Au—Pt bi-metallic nanocrystals dried from the nanocrystal suspensions.
- FIG. 30 contains the UV-Vis data collected for this sample (GPB-040), specifically displaying the 350-900 nm range.
- GPB-040 concentrated samples were prepared via Tangential Flow Filtration (TFF), as described herein where the diafiltration buffer was substituted with de-ionized water to remove the process enhancer from the solution.
- GPB-040 was concentrated 20 fold by volume three times, each time reconstituting with de-ionized water. Subsequently, TFF concentrated GPB-040 was then centrifuged at 11,000 rpm for 10 minutes resulting in the presence of a Au—Pt bi-metallic pellet at the bottom of a 1.5 mL centrifuge tube. Approximately 24 tubes were used to collect a final sample of about 1.5 mL with a concentration that is about 400 times greater than the starting solution. This solution was then deposited onto the sample stub as discussed below.
- TFF Tangential Flow Filtration
- TFF tangential flow filtration
- a feed tank 1001 provides fluid to a feed pump 1002 and into a filtration module 1003 .
- the filtrate stream 1004 is discarded.
- Retentate is diverted through the retentate valve 1005 and returned as 1006 into the feed tank 1001 .
- the applied pressure forces a portion of the fluid through the membrane and into the filtrate stream, 1004 .
- Any particulates and macromolecules that are too large to pass through the membrane pores are retained on the upper stream and swept along by the tangential flow into the retentate, 1006 .
- the retentate, having a higher concentration of colloidal particles is returned back to the feed tank, 1001 . If there is no diafiltration buffer added to the feed tank, then the colloid volume in the feed tank, 1001 , is reduced by the amount of filtrate removed and the suspension becomes concentrated.
- Millipore Pellicon XL cassettes were used with 5 kDa and 10 kDa MWCO cellulose membranes.
- the retentate pressure was set to 40 PSI by a retentate valve, 1005.
- 10 kDa membrane allows approximately 4 times higher filtrate flow rate related to a 5 kDa membrane under the same transmembrane pressure, which is expected for a larger pore size.
- pores of 10 kDa membrane are small enough to retain all formed bi-metallic nanocrystals in the retentate in GPB-040.
- XPS X-ray photoelectron spectroscopy
- Spectra were collected within two ranges, (i.e., a low resolution survey scan and a higher resolution multiplex scan in specific regions of interest). Survey scans were taken between binding energies of 0-1200 eV while higher resolution scans were taken between 80-100 eV and 65-85 eV. Elemental gold exhibits a multiplet (4f 512 & 4f 712 ) at 87.6 eV and 83.9 eV, respectively, and information such as oxide composition and concentration can be determined from the expanded region at 80-100 eV. Platinum exhibits a multiplet (4f 512 & 4f 712 ) at 74.5 eV and 71.2 eV, respectively, and information such as concentration and oxide content can be determined from the expanded region at 65-85 eV.
- Sputter cleaning and depth profiling were carried out with a Sputter Ion Gun, (PHI, Model 04-303).
- the incident ion gun was operated at an accelerating voltage of 4.0 keV, and sample currents were maintained at about 25 mA across the sample area.
- the pressure in the main chamber was maintained at about 5 ⁇ 10 ⁇ 8 Torr.
- the corresponding raster size is 4 ⁇ 4 mm with a pressure of 25 mPa.
- Sputtering was done at intervals of 5, 10, 20, 30, 40, 50, 70, 90, 120, 180, & 240 minutes.
- FIGS. 29 h - 29 i are spectra collected from GPB-040, a gold-platinum bi-metallic nanocrystal suspension.
- the spectra were prepared by placing 100-200 uL of sample onto the sample stub and subsequently pulling a vacuum to dry the material onto the carbon tape. The chamber was then opened and another 100-200 uL was deposited. This process was repeated eleven times to produce a thin film of material on the carbon tape.
- the initial survey scan, FIG. 29 h is useful in determining surface contaminants and elemental composition of the nanocrystals.
- peaks indicative of carbon, oxygen, platinum, and gold are peaks indicative of carbon, oxygen, platinum, and gold.
- the small carbon peak at 285 eV is from incomplete sample coverage of the carbon tape while the oxygen peak at 531 eV is likely a result of trapped oxygen due to the sample preparation technique; however in a layer of adsorbed oxygen may have become trapped in between drop depositions. Peaks at 690 eV and 750 eV can be attributed to fluorine sample chamber contamination and oxygen, respectively. In both instances the peaks disappeared after a 30 minute sputter.
- FIG. 29 i Higher resolution multiplex scans, FIG. 29 i , between 60 eV-100 eV provide additional information on the gold and platinum composition of the nanocrystals.
- the Au 4f 512 peak at 88 eV contains a small shoulder that can be attributed to sample charging. After a 30 minute sputter, the flow of positive argon ions neutralized the sample and the shoulder disappeared. In addition, the Pt 4f 7/2 peak rises after the 30 minute sputter at about 71 eV.
- FIGS. 29 a - g Au—Pt bi-metallic nanocrystal solutions are heterogeneous in structure with respect to atomic platinum and atomic gold.
- energy dispersive spectra EDS
- FIGS. 29 b and 29 d Resultant EDS data is displayed in FIGS. 29 b and 29 d .
- a platinum peak at about 9.4 keV and a gold peak at about 9.7 keV are present.
- FIGS. 29 e - g are Scanning Transmission Electron Microscopy (STEM) images of bi-metallic nanocrystals from suspension GPB-040.
- FIG. 29 e is a STEM image of at least four Au—Pt bi-metallic nanocrystals dried on a copper grid.
- FIGS. 29 f and g are platinum and gold EDS mappings, respectively, of the nanocrystals imaged in FIG. 29 e . It is clear from FIGS. 29 f and 29 g that both platinum and gold exist heterogeneously throughout the examined nanocrystals.
- FIGS. 29 h and 29 i provide further evidence that the nanocrystal surfaces are both free from organic contamination and do not exhibit a core-shell behavior.
- a dialysis bag technique permits the gradual concentration of colloids made according to the teachings herein. Colloidal suspensions were placed inside of a dialysis bag and the bag itself was immersed into an aqueous solution of a PEG-based polymer, which creates a negative osmotic pressure. The negative osmotic pressure resulted in the extraction of water from the colloid maintained within (i.e., inside) the dialysis bag.
- FIG. 31 a shows a dialysis bag 2000 , containing a representative colloid suspensions 3000 .
- a suitable plastic container 5000 (made of HDPE plastic) and a PEG-based polymer material 1000 therein.
- the dialysis membrane which forms the dialysis bag 2000 , is characterized by molecular weight cut off (MWCO)—an approximate achieved threshold size above which larger-sized species will be retained inside of the membrane.
- MWCO molecular weight cut off
- Dialysis concentration was achieved by using a cellulose membrane having a 3.5 kDa MWCO for the dialysis bag 2000 and the polymer solution 1000 was made from a PEG-8000 polymer. Under these conditions, water molecules and small ions could pass through the dialysis membrane of the bag 2000 , but colloidal nanoparticles larger than the 3.5 kDa MWCO would be retained inside the dialysis bag. However, PEG-8000 molecules cannot pass through (i.e., due to their size) the membrane and remained outside of the dialysis bag 2000 .
- FIG. 31 b shows that the dialysis bag 2000 shrank in volume (over time) relative to its size in FIG. 31 a .
- the dialysis bag 2000 should not be allowed to collapse as liquid is removed from the bag.
- nanocrystals that may remain on the inner surface of the bag should not be over-stressed so as to prevent their possible aggregation.
- Each dialysis bag 2000 was filled with approximately 400 to 500 mL of nanocrystal suspension 3000 , and maintained in the PEG-8000 solution 1000 until the bag volume was reduced approximately 10 times in size and volume. Further suspension concentration, if required, occurred by combining 10 ⁇ concentrated colloids from several bags into one bag and repeating the same set of concentration steps again. Dialysis bags 2000 can safely be used about 10 times without achieving any noticeable membrane fouling.
- the starting PEG-8000 concentration 1000 in the polymer solution outside the dialysis bag 2000 was about 250 g/L and was naturally lowered in concentration due to water being drawn out from the colloid 3000 through the dialysis bags 2000 (i.e., due to the created osmotic pressure). Higher polymer concentrations and gentle stirring can increase the rate of water removal from the colloid 3000 .
- This dialysis process concentrated the gold colloids with no visible staining of the dialysis bags 2000 .
- the concentration of remaining gold nanocrystals in suspension 4000 was estimated by volume reduction and also measured by ICP-MS techniques (discussed in detail later herein).
- the remaining gold in the suspension 4000 was similar to the gold concentration measured directly by ICP-MS techniques.
- part of the platinum produced in the first electrochemical step was ionic, and some amount of this ionic form of platinum removal after the second electrochemical processing steps and passed through the dialysis bag 2000 during concentration. This effect resulted in a lower concentration factor for atomic platinum relative to atomic gold (all of the atomic gold was apparently in metallic form).
- the Au—Pt bi-metallic nanocrystal suspension slightly stained the membrane of the dialysis bag 2000 to a yellowish-green uniform color.
- the dialysis bag technique was used to achieve a series of concentration ranges of two different colloidal suspensions that were used in a subsequent in-vitro cellular culture experiment.
- Table 24 sets forth 9 different concentrations of metals in a formed gold suspension (NE10214) and in an Au/Pt bi-metallic suspension (GPB-032) the formations of which are described earlier herein. Concentration values were measured by inductively coupled plasma-mass spectrometry (ICP-MS) as desribed immediately below.
- ICP-MS inductively coupled plasma-mass spectrometry
- ICP-MS Inductively Coupled Plasma-Mass Spectometry
- the ICP-MS values were obtained from an Agilent 7700x
- the technique of inductively coupled plasma spectroscopy-mass spectrometry requires a liquid sample to be introduced into a sample chamber via a nebulizer, thus removing the larger droplets, and introducing a fine aerosol spray into the torch chamber carried via a supply of inert Argon gas.
- the torch temperature ranges between 8000K-10000K.
- the aerosol is instantly desolvated and ionized within the plasma and extracted into the first vacuum stage via the sampling cone and then subsequently passes through a second orifice, the skimmer cone.
- the ions are then collimated by the lens system and then focused by the ion optics.
- the ion lenses allow the ICP-MS to achieve high signal sensitivity by preventing photons and neutral species from reaching the detector by mounting the quadrupole and detector off axis from the entering ion beam.
- the cell gas, Helium is introduced into the ORS which is an octopole ion guide positioned between the ion lens assembly and the quadrupole. Interferences such as polyatomic species are removed via kinetic energy discrimination.
- the ions that pass through then proceed into the quadrupole mass analyzer which consists of four long metal rods. RF and DC voltages are applied at the rods and it is the variation in voltages that allow the rods to filter ions of specific mass-to-charge ratios. The ions are then measured by the pulse analog detector.
- an ion When an ion enters the electron multiplier, it strikes a dynode and creates an abundance of free electrons which then strike the next dynode, resulting in the creation of additional electrons.
- the amount of ions from a specific element correlates to the amount of electrons generated, thus resulting in more or less counts, or CPS.
- Samples were prepared by diluting 500 ⁇ L of sample in 4.5 mL of 5% HNO 3 /2% HCl for 30 minutes at 70° C. Samples were prepared in triplicate. Subsequently, samples were transferred to a polypropylene test tube which was then placed in a rack in the Cetac autosampler.
- the Agilent ICP-MS 7700x plasma was turned on and a start up procedure was initialized. The plasma was allowed to warm up for 26 minutes prior to running the initial optimization. After successful completion of the optimization steps, the instrument was then ready for analysis. A quick manual tune was performed and the signal of low, mid, and high masses (59, 89, & 205) were checked to ensure that the instrument was within our internal specifications. Afterwards, the internal standard line tubing was switched from a 5% HNO 3 blank to an internal standard solution containing In 115.
- Calibration samples and independent continuous concentration verification (ICCV) standards were prepared from external stock solutions prepared by SPEX CertiPrep. Multi-Element 3 calibration standards containing gold were serially diluted from 10 ppm to 1000 ppb, 100 ppb, 10 ppb, and 1 ppb, respectively. A blank solution of the diluent, 5% HNO 3 /2% HCl, was used as the 0 ppb standard.
- the ICCV sample was placed in a sample vial and placed on a rack with the calibration standards. Prior to sample analysis, a calibration curve was created by measuring 0 ppb, 1 ppb, 10 ppb, 100 ppb, & 1000 ppb. Samples of interest were then measured with a 90 second 5% HNO 3 rinse step in between sample uptake. After every 6 samples, the ICCV was run to ensure that the calibration curve was within 10% of the actual values.
- a cell line panel was assembled with 30 different human tumor types selected from the ATCC and DSMZ (all DSMZ cell lines are marked with “*”) culture banks and included typical bladder, breast, cervix, CNS, colon, H&N, lung, ovary, prostate, stomach, thyroid, uterus and vulva cancers.
- the 30 specific cell lines and tumor types are set forth in Table 25.
- Cells were grown in RPMI1640, 10% FBS, 2 mM L-alanyl-L-Glutamine, 1 mM Na Pyruvate in a humidified atmosphere of 5% CO 2 at 37° C. Cells were seeded into 384-well plates and incubated in a humidified atmosphere of 5% CO 2 at 37° C. Compounds NE10214 and GPB-032 were added 24 hours post cell seeding. At the same time, a time zero untreated cell plate was generated.
- cells were fixed and stained with fluorescently labeled antibodies and nuclear dye to allow visualization of nuclei, apoptotic cells and mitotic cells.
- Apoptotic cells were detected using an anti-active caspase-3 antibody.
- Mitotic cells were detected using an anti phospho-histone-3 antibody.
- the concentrated Au suspension (NE10214, also “Compound 1”) and the concentrated bi-metallic suspension AuPt (GPB-032, also “Compound 2”) were diluted as shown in Table 26 below and assayed over 9 concentrations from the highest test concentration to the lowest test concentration. When the two test compounds were added to the growth medium they became diluted by the growth media.
- the actual atomic concentrations of the metallic components (i.e., Au in NE10214; and Au+Pt in GPB-032) in the growth media are shown in Table 26 as “In Vitro Conc microM”.
- the multiplexed cytotoxicity assay used a cell image based analysis technique where cells were fixed and stained with fluorescently labeled antibodies and nuclear dye as mentioned above.
- Cell proliferation was measured by the signal intensity of the incorporated nuclear dye.
- the cell proliferation assay output is referred to as the relative cell count.
- the cell proliferation data output was transformed to percent of control (POC) using the following formula:
- Relative cell count IC 50 is the test compound concentration at 50% of maximal possible response.
- a relative cell count EC 50 is the test compound concentration at the curve inflection point or half the effective response (parameter C of the fitted curve solution).
- GI 50 is the concentration needed to reduce the observed growth by half. This is the concentration that inhibits the growth midway between untreated cells and the number of cells seeded in the well (Time zero value).
- the output of each biomarker is fold increase over vehicle background normalized to the relative cell count in each well.
- the activated caspase-3 marker labels cells from early to late stage apoptosis.
- the output is shown as a fold increase of apoptotic cells over vehicle background normalized to the relative cell count in each well. Concentrations of test compound that cause a 5-fold induction in the caspase-3 signal indicates significant apoptosis induction. Wells with concentrations higher than the relative cell count IC 95 are eliminated from the caspase3 induction analysis.
- the phospho-histone-3 marker labels mitotic cells.
- the output is shown as a fold induction of mitotic cells over vehicle background normalized to the relative cell count in each well. When the fold induction of mitotic cell signal over background is ⁇ 1, there is “no effect” on the cell cycle. Two or more fold increase in phospho-histone-3 signal over vehicle background indicates significant test compound induction of mitotic block.
- Two or more fold decrease in the phospho-histone-3 signal may indicate G1/S block only when cytotoxicity levels are below the measured relative cell count IC 95 .
- the decrease in mitotic cell counts are most likely due to a more general cytotoxicity effect rather than a true G1/S phase block.
- Wells with concentrations higher than the relative cell count IC 95 are eliminated from the phospho-histone-3 analysis.
- the concentrated Au suspension and the concentrated Au—Pt bi-metallic suspension show distinctly different patterns of the presence of anti-cancer activity, and distinctly different patterns of the type of anti-cancer activity, across the thirty different tumor cell lines.
- FIGS. 32 a - 32 ad show graphically the difference in performance of compound 1 and compound 2 against each of the 30 cell lines tested. Specifically, comparisons are set forth for each of “Relative Cell Count %”, “Apoptosis (fold induction)” and “Mitosis (fold induction)”. The data show that there is a significant elevation in apoptosis induction in eight different tumor cell lines treated with the concentrated Au—Pt bi-metallic suspension (GPB-032), but this kind of activity is not shown in any of the tumor cell lines treated with the concentrated Au compound (NE10214).
- GPB-032 concentrated Au—Pt bi-metallic suspension
- the concentrated Au—Pt bi-metallic suspension shows significant anti-cancer activity in twelve tumor cell lines where the concentrated Au compound showed no activity at all, and the concentrated Au suspension is effective in two additional tumor cell lines where the concentrated AuPt bi-metallic suspension shows no activity at all,—so in fourteen of thirty tumor cell lines, there is no shown overlap in the presence of any kind of anti-cancer activity.
- This Example demonstrates the efficacy of several orally administered inventive compositions in a mouse xenograft cancer model.
- Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein.
- the Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm 3 in size.
- the Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm 3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle.
- the recipient mice were randomly placed into treatment groups, 3 per group and the oral treatment was started. Treatment was given exclusively via the drinking bottle shared between 3 mice in each group. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated at day 24.
- the results of the Example are summarized in FIGS. 33 a - 33 b.
- GB-218 was prepared similarly to Example 1 resulting in a gold concentration of 7.6 ppm as measured by AAS. Additionally said solution was determined to have a hydrodynamic radius of 15.1 nm as measured by the Viscotek.
- GB-219 was prepared similarly in regards to Example 1 wherein potassium hydroxide was replaced as the process enhancer for sodium bicarbonate at a concentration of 0.63 g/gallon (i.e., about 0.17 mg/mL). GB-219 had a gold concentration of 8.7 ppm as measured by AAS. Additionally said solution was determined to have a hydrodynamic radius of 18.3 nm as measured by the Viscotek.
- PB-39 was prepared similarly to Example 13 PB57001 example, resulting in a suspension of nanocyrystal platinum particles having a Pt concentration of 7.4 ppm.
- PB-22-C4 was prepared similarly to Example 13, wherein the applied frequency of 501 AC was set to 80 Hz instead of 5 Hz to produce a solution comprising predominantly of Pt ionic species with a small amount of Pt nanocrystalline species.
- the concentration of sodium bicarbonate was 2.5 g/gallon (i.e., about 0.66 mg/mL).
- PB-22-C4 was then subsequently concentrated using an electrical hot plate to produce a Pt concentration of about 8.3 ppm.
- HCT 116 cell line ATCC CCL-247.
- PBS Phosphate buffered saline
- Test compounds platinum nanocrystal suspension, gold nanocrystal suspension and Au—Pt bi-metallic suspension.
- Positive control compound cisplatin.
- Negative control compound drinking water.
- Negative Control Group 1 Days 0-24, given normal drinking water.
- Positive Control Group 2 Days 0-24, given normal drinking water; and given a daily cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”).
- Treatment Group 3-6 Days 0-24, given test compounds as their drinking water.
- Protocol A Preparation and Growth of Donor Tumors
- the cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer. 5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 ⁇ L contained about 3 ⁇ 10 6 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site.
- mice had previously arrived and their health was checked. 2. All animals were allowed to acclimate for at least 72 hours. 3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation. 4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe. 5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm 3 . 6.
- Protocol B Insertion of Tumors from Donor Mice into Recipient Mice 1. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag. 2. The recipient mice were allowed to acclimate for at least 72 hours. 3. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm 3 in size.
- the 2 mm 3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank).
- the tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm 3 before treatment started at day 0.
- Treatments continued for 24 days or until the mouse was removed from the study and euthanized or the mouse died. 4.
- the tumor sizes and weights of the animals were determined daily until the end of the study at day 24.
- FIGS. 33 a and 33 b show graphically the results of the oral test.
- FIG. 33 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better.
- FIG. 33 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better.
- Table 29 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia.
- the Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
- This Example demonstrates the efficacy of several intratumorally (“IT”) administered inventive metallic nanocrystal suspensions in a mouse xenograft cancer model.
- Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein.
- the Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm 3 in size.
- the Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm 3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle.
- the recipient mice were randomly placed into treatment groups, 3 per group and the “IT” treatment was started. Treatment was given exclusively by needle injection into the tumor twice a day. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated at day 30.
- the results of the Example are summarized in FIG. 34 a - 34 b.
- GB-218 was prepared similarly to Example 1 resulting in a gold concentration of 7.6 ppm as measured by AAS. Additionally said solution was determined to have a hydrodynamic radius of 15.1 nm as measured by the Viscotek.
- GB-219 was prepared similarly in regards to Example 1 wherein potassium hydroxide was replaced as the process enhancer for sodium bicarbonate at a concentration of 0.63 g/gallon (i.e., about 0.17 mg/mL). GB-219 had a gold concentration of 8.7 ppm as measured by AAS. Additionally said solution was determined to have a hydrodynamic radius of 18.3 nm as measured by the Viscotek.
- PB-39 was prepared similarly to Example 13 PB57001 example, resulting in a suspension of nanocyrystal platinum particles having a Pt concentration of 7.4 ppm.
- PB-22-C4 was prepared similarly to Example 13, wherein the applied frequency of 501 AC was set to 80 Hz instead of 5 Hz to produce a solution comprising predominantly of Pt ionic species with a small amount of Pt nanocrystalline species.
- the concentration of sodium bicarbonate was 2.5 g/gallon (i.e., about 0.66 mg/mL).
- PB-22-C4 was then subsequently concentrated using an electrical hot plate to produce a Pt concentration of about 8.3 ppm.
- HCT 116 cell line ATCC CCL-247.
- PBS Phosphate buffered saline
- Test compounds platinum nanocrystal suspension, gold nanocrystal suspension and Au—Pt bi-metallic suspension.
- Positive control compound cisplatin.
- Negative control compound drinking water.
- Negative Control Group 1 Days 0-30, saline injection twice a day, with a total of 100 ⁇ l in each tumor divided between 2-3 injection points; (given normal drinking water to drink).
- Positive Control Group 2 Days 0-30, cisplatin injection 8 mg/kg given once a day into the peritoneum (IP) (given normal drinking water to drink).
- Treatment Group 3-6 Days 0-30, nanocrystal formulation injection twice a day, with a total of 100 ⁇ l in each tumor divided between 2-3 injection points; (given normal drinking water to drink).
- Protocol A Preparation and Growth of Donor Tumors
- the cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer. 5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 ⁇ L contained about 3 ⁇ 10 6 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site.
- mice had previously arrived and their health was checked. 2. All animals were allowed to acclimate for at least 72 hours. 3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation. 4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe. 5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm 3 . 6.
- Protocol B Insertion of Tumors from Donor Mice into Recipient Mice 5. Additional Blab/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag. 6. The mice were allowed to acclimate for at least 72 hours. 7. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm 3 in size.
- the 2 mm 3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank).
- the tumors were permitted to grow in the recipient mice until they reached a size of about 7 ⁇ 7 mm before treatment started at day 0. Treatments continued for 30 days or until the mouse was removed from the study and euthanized or the mouse died. 8. The tumor sizes and weights of the animals were determined daily until the end of the study at day 24.
- Protocol C Intertumoral Injection into Recipient Mice 1.
- Each tumor in each recipient mouse was injected twice daily (about 12 hours apart) with about 100 ⁇ l of either negative control, positive control or test compound.
- the needle used for injection was either a 25 Ga or 26 Ga needle. Depending on the tumor size, there were either 2 or 3 injection points for each tumor.
- FIGS. 34 a and 34 b shows graphically the results of the IT test.
- FIG. 34 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better.
- FIG. 34 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better.
- Table 30 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia.
- the Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
- This Example demonstrates the relative efficacy of four orally administered inventive metallic nanocrystal suspensions in a mouse xenograft cancer model.
- Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein.
- the Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm 3 in size.
- the Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm 3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle.
- mice were randomly placed into treatment groups, 6 per group and the oral treatment was started.
- 6 mice were in the positive control group (“Cisplatin”) and 6 mice were in the negative control group and received only water (“Control”).
- Treatment was given exclusively via the drinking bottle shared between the mice in each Treatment group.
- Cisplatin was given by intraperitoneal injection on day 0. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated as scheduled.
- the results of the Example are summarized in FIGS. 35 a - 35 b .
- HCT 116 cell line ATCC CCL-247.
- PBS Phosphate buffered saline
- Test compounds platinum nanocrystal suspension, gold nanocrystal suspension and Au—Pt bi-metallic suspension.
- Positive control compound cisplatin.
- Negative control compound drinking water.
- Negative Control Group 1 Days 0-24, given normal drinking water.
- Positive Control Group 2 Days 0-24, given normal drinking water; and given a one-time cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”) on day 0.
- Treatment Group 3-6 Days 0-24, given test compounds as their drinking water.
- Protocol A Preparation and Growth of Donor Tumors
- the cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer. 5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 ⁇ L contained about 3 ⁇ 10 6 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site.
- mice had previously arrived and their health was checked. 2. All animals were allowed to acclimate for at least 72 hours. 3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation. 4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe. 5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm 3 . 6.
- Protocol B Insertion of Tumors from Donor Mice into Recipient Mice 9. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag. 10. The recipient mice were allowed to acclimate for at least 72 hours. 11. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm 3 in size.
- the 2 mm 3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank).
- the tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm 3 before treatment started at day 0. Treatments continued for 24 days or until the mouse was removed from the study and euthanized or the mouse died. 12. The tumor sizes and weights of the animals were determined daily until the end of the study at day 24.
- FIGS. 35 a and 35 b show graphically the results of the oral test.
- FIG. 35 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better.
- FIG. 35 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better.
- Table 31 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia.
- the Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
- Table 32 provides a comparison of the doubling time (RTV2) for each group in the study. In addition, table 32 also lists the growth delay in days, maximum percent weight loss and statistical significance of the data.
- This Example demonstrates the relative efficacy of three orally administered inventive metallic nanocrystal suspensions in a mouse xenograft cancer model relative to Cisplatin.
- Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein.
- the Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm 3 in size.
- the Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm 3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle.
- mice were randomly placed into treatment groups, 8 per group and the oral treatment was started. 8 mice were in the positive control group (“Cisplatin”) and 8 mice were in the negative control group and received only water (“Control”). Treatment was given exclusively via the drinking bottle shared between the mice in each Treatment group. Cisplatin was given by intraperitoneal injection on day 0. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated as scheduled. The results of the Example are summarized in FIGS. 36 a - 36 b .
- HCT 116 cell line ATCC CCL-247.
- PBS Phosphate buffered saline
- Test compounds Au—Pt bi-metallic nanocrystal suspensions.
- Positive control compound cisplatin.
- Negative control compound drinking water.
- Negative Control Group 1 Days 0-21, given normal drinking water.
- Positive Control Group 2 Days 0-21, given normal drinking water; and given a one-time cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”) on day 0.
- Treatment Group 3-5 Days 0-21, given test compounds as their drinking water.
- Protocol A Preparation and Growth of Donor Tumors
- the cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer. 5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 ⁇ L contained about 3 ⁇ 10 6 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site.
- mice had previously arrived and their health was checked. 2. All animals were allowed to acclimate for at least 72 hours. 3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation. 4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe. 5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm 3 . 6.
- Protocol B Insertion of Tumors from Donor Mice into Recipient Mice 13. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag. 14. The recipient mice were allowed to acclimate for at least 72 hours. 15. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm 3 in size.
- the 2 mm 3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank).
- the tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm 3 before treatment started at day 0.
- Treatments continued for 21 days or until the mouse was removed from the study and euthanized or the mouse died. 16.
- the tumor sizes and weights of the animals were determined daily until the end of the study at day 21.
- FIGS. 36 a and 36 b show graphically the results of the oral test.
- FIG. 36 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better.
- FIG. 36 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better.
- Table 33 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia.
- the Sample IDs relate to compounds manufactured according to procedures discussed earlier herein.
- This Example demonstrates the relative efficacy of three orally administered inventive Au—Pt bi-metallic nanoparticle suspensions in a mouse xenograft cancer model relative to Cisplatin.
- Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein.
- the Balb/C donor mice were used to grow H460 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm 3 in size.
- the Balb/C recipient mice were given brief general anesthesia and then one H4602 mm 3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle.
- mice were randomly placed into treatment groups, 8 per group and the oral treatment was started. 8 mice were in the positive control group (“Cisplatin”) and 8 mice were in the negative control group and received only water (“Control”). Treatment was given exclusively via the drinking bottle shared between the mice in each Treatment group. Cisplatin was given by intraperitoneal injection on day 0. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated as scheduled. The results of the Example are summarized in FIGS. 37 a - 37 b .
- H460 cell line ATCC HTB-177.
- Phosphate buffered saline PBS
- Test compounds Au—Pt bi-metallic nanocrystal suspensions.
- Positive control compound cisplatin.
- Negative control compound drinking water.
- Negative Control Group 1 Days 0-21, given normal drinking water.
- Positive Control Group 2 Days 0-21, given normal drinking water; and given a one-time cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”) on day 0.
- Treatment Group 3-5 Days 0-21, given test compounds as their drinking water.
- Protocol A Preparation and Growth of Donor Tumors
- the cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer. 5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 ⁇ L contained about 3 ⁇ 10 6 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site.
- mice had previously arrived and their health was checked. 2. All animals were allowed to acclimate for at least 72 hours. 3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation. 4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe. 5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm 3 . 6.
- Protocol B Insertion of Tumors from Donor Mice into Recipient Mice 17. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag. 18. The recipient mice were allowed to acclimate for at least 72 hours. 19. H460 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm 3 in size.
- the 2 mm 3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank).
- the tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm 3 before treatment started at day 0. Treatments continued for 24 days or until the mouse was removed from the study and euthanized or the mouse died. 20.
- the tumor sizes and weights of the animals were determined daily until the end of the study at day 21.
- FIGS. 37 a and 37 b show graphically the results of the oral test.
- FIG. 37 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better.
- FIG. 37 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better.
- Table 35 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia.
- the Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
- This Example demonstrates the relative efficacy of one orally administered inventive Au—Pt bi-metallic nanocrystalline suspension in a mouse xenograft cancer model.
- Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein.
- the Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm 3 in size.
- the Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm 3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle.
- mice were randomly placed into treatment groups, 8 per group and the oral treatment was started. 8 mice were in the positive control group (“Cisplatin”) and 8 mice were in the negative control group and received only water (“Control”). Treatment was given exclusively via the drinking bottle shared between the mice in each Treatment group. Cisplatin was given by intraperitoneal injection on day 0. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated as scheduled. The results of the Example are summarized in FIGS. 38 a - 38 b
- HCT 116 cell line ATCC CCL-247. Phosphate buffered saline (“PBS”). Test compounds: gold nanocrystal suspension NE-28-10X (NE-28 produced equivalent to
- Positive control compound cisplatin.
- Negative control compound drinking water.
- Negative Control Group 1 Days 0-21, given normal drinking water.
- Positive Control Group 2 Days 0-21, given normal drinking water; and given a one-time cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”) on day 0.
- Treatment Group 3 Days 0-21, given test compounds as their drinking water.
- Protocol A Preparation and Growth of Donor Tumors
- mice had previously arrived and their health was checked. 2. All animals were allowed to acclimate for at least 72 hours. 3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation. 4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26-gauge needle was subsequently added to the syringe. 5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm 3 . 6.
- Protocol B Insertion of Tumors from Donor Mice into Recipient Mice 21. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag. 22. The recipient mice were allowed to acclimate for at least 72 hours. 23. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm 3 in size.
- FIGS. 38 a and 38 b show graphically the results of the oral test.
- FIG. 38 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better.
- FIG. 38 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better.
- Table 37 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia.
- the Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
- Test gold/platinum bi-metallic nanocrystalline suspensions GPB-15-1, GPB-15-2 and GPB-030-01 (equivalent to PGT24).
- GPB-15-1 Average 4.0 ml 1 d; gold ppm: 8.6. platinum ppm: 2.3) as drinking water from day 0-day 47.
- GPB-15-2 (average 3.9 ml 1 d; gold ppm: 8.6: platinum ppm: 2.3) as drinking water from day 48-day 101.
- GPB-030-01 average 4.3 ml 1 d; gold ppm: 8.6, platinum ppm: 2.5) as drinking water from day 102 through 39 weeks.
- the animals were allowed to acclimate for at least 1 week.
- Gold/platinum bi-metallic nanocrystalline suspension were prepared so as to achieve a suspension with a concentration of about 8.6 ppm Au and 2.3 ppm Pt for GPB-15-1, 8.6 ppm Au and 2.3 ppm Pt for GPB-15-2 and 8.6 ppm Au and 2.5 ppm Pt in GPB-030-01.
- Animals were housed in a 10-gallon glass tank with a metal mesh cover.
- a corn cob bedding material (Bed O′ Cobs manufactured by the Andersons) was provided as a floor material, one nestlet (purchased from Ancare) was given per animal per week.
- Animals had access to a wheel for exercise (8 in diameter Run around wheel manufactured by Super Pet), as well as a housing unit (Pet igloo by Super Pet) and a plastic food dish (Petco plastic dish) for Certified Rodent diet.
- FIG. 39 b shows the average weight gain of Treatment Group 2 and Control Group 1.
- Albumin is a known stabilizing agent and could provide a biofunctionalized layer for water-dispersed nanoparticles.
- the binding affinity between gold nanoparticles and DNA has been indicated to affect DNA transcription.
- Albumin is also known to assist in drug delivery.
- Albumin was incubated with GPB-11 in a binding buffer at room temperature for about 1 hour to determine the differential binding of albumin to GPB-11 in the absence or presence of genomic DNA.
- genomic DNA was incubated with GPB-11 for about 1 hour to measure the binding abilities of DNA to the GPB-11 when co-incubating with or without albumin. After reactions were allowed to occur, the GPB-11 suspension was spun down, washed and placed into an elution buffer for absorbance measurements.
- AFM Fast-scan atomic force microscopy
- the concentrated GPB-11 suspension was rehydrated in a solution containing 2.7 mM Sodium Hydrogen Carbonate and 2.1 mM Potassium hydroxide with the same amount as the above-described supernatant.
- Zeta potentials of rehydared GPB-11 and original GPB-11 solutions were measured using a Zetasizer as discusssed elswhere herein, and the results were ⁇ 50.3 mV and ⁇ 51.7 mV respectively.
- the very similar Zeta potential values suggested that rehydration of concentrated GPB-11 in the binding reaction studies should have the same effect as adding an oringinal concentration of GPB-11.
- the binding buffer was prepared with 20 mM Tris, 100 mM KCl, 3 mM MgCl2 and 0.1% IGEPAL.
- the pH was adjusted to about 7.5 by pH/Conductivity Meter with Hydrochloric acid and NaOH.
- the absorbance of albumin binding to GPB-11 was measured at 280 nm. Different combinations of albumin and GPB-11 were tested in the presence or absence of genomic DNA. Table 30 shows that very similar results were achieved among different albumin and GPB11 combinations. Representative data are also depicted in FIG. 40 a .
- FIG. 40 a shows graphically the amount of mouse albumin binding in the presence or absence of mouse genomic DNA as a function of the absorbance at 280 nm.
- albumin significantly bound to the bi-metallic nanocrystals in GPB-11.
- no albumin binding to the nanocrystals in GPB-11 was observed.
- the nanocrystals in GPB-11 can bind with albumin, but preferentially binds to mouse genome DNA.
- the Au—Pt bi-metallic nanocrystals in GPB-11 apparently have a soft corona of albumin.
- DNA binding to nanocrystals in GPB-11 was determined by measuring the absorbance at 260 nm.
- the binding ability of mouse or human genomic DNA to bi-metallic nanocrystals in GPB-11 was measured with different combinations of albumin.
- Table 31 shows the various combinations or mixtures tested. Highly consistent results were observed between different DNA and nanocrystals in GPB-11 combinations. The representative results are depicted graphically in FIG. 40 b .
- FIG. 40 b shows graphically the amount of DNA binding in the presence or absence of mouse albumin.
- FIG. 40 b shows that in both, the presence and the absence of albumin, genomic DNA significantly bound to nanocrystals in GPB-11.
- albumin was absent, the amount of DNA binding with GPB-11 nanocrystals was dramatic.
- a statistically significant amount of DNA was observed to be bound to the GPB-11 bi-metallic nanocrystals.
- These results further confirm that bi-metallic nanocrystals in GPB-11 bind to genomic DNA much stronger than albumin.
- the Au—Pt bi-metallic nanocrystals in GPB-11 may bind to genomic DNA (when in the presence thereof) with covalent bonds. Such bonding could affect DNA function.
Abstract
The present invention relates to novel gold-platinum based bi-metallic nanocrystal suspensions that have nanocrystal surfaces that are substantially free from organic or other impurities or films associated with typical chemical reductants/stabilizers and/or raw materials used in nanoparticle formation processes. Specifically, the surfaces are “clean” relative to the surfaces of metal-based nanoparticles made using chemical reduction (and other) processes that require organic (or other) reductants and/or surfactants to grow (and/or suspend) metal nanoparticles from metal ions in a solution.The invention includes novel electrochemical manufacturing apparatuses and techniques for making the bi-metallic nanocrystal suspensions. The techniques do not require the use or presence of chlorine ions/atoms and/or chlorides or chlorine-based materials for the manufacturing process/final suspension. The invention further includes pharmaceutical compositions thereof and the use of the bi-metallic nanocrystals or suspensions or colloids thereof for the treatment or prevention of diseases or conditions for which metal-based therapy is already known, including, for example, for cancerous diseases or conditions.
Description
- The present application is a divisional of U.S. application Ser. No. 15/204,534, filed Jul. 7, 2016. U.S. application Ser. No. 15/204,534, is a divisional of U.S. application Ser. No. 14/008,931, filed on Dec. 16, 2013; now U.S. Pat. No. 9,387,225, issued Jul. 12, 2016. The aforementioned parent application is a U.S. national phase entry of International Application No. PCT/US2012/031654 filed on Mar. 30, 2012, which claims priority to U.S. 61/469,525, filed on Mar. 30, 2011. All of the aforementioned applications are hereby expressly incorporated by reference.
- The present invention relates to novel gold-platinum based bi-metallic nanocrystal suspensions that have nanocrystal surfaces that are substantially free from organic or other impurities or films associated with typical chemical reductants/stabilizers and/or raw materials used in nanoparticle formation processes. Specifically, the surfaces are “clean” relative to the surfaces of metal-based nanoparticles made using chemical reduction (and other) processes that require organic (or other) reductants and/or surfactants to grow (and/or suspend) metal nanoparticles from metal ions in a solution.
- The invention includes novel electrochemical manufacturing apparatuses and techniques for making the bi-metallic nanocrystal suspensions. The techniques do not require the use or presence of chlorine ions/atoms and/or chlorides or chlorine-based materials for the manufacturing process/final suspension. The invention further includes pharmaceutical compositions thereof and the use of the bi-metallic nanocrystals or suspensions or colloids thereof for the treatment or prevention of diseases or conditions for which metal-based therapy is already known, including, for example, for cancerous diseases or conditions.
- One motivation for making metallic-based nanoparticles is the novel performance achieved at the nano-scale relative to bulk materials. Materials of nanoscopic dimensions offer a variety of different properties than those observed on the macroscale, thus potentially enabling a variety of unique applications. In particular, nanometals exhibit a variety of electronic, optical, magnetic and/or chemical properties which are typically not achievable when metallic materials are in their bulk form. For example, metals that are relatively inert at the macroscale, such as platinum and gold, are excellent catalysts at the nanoscale. Further, combinations of two different metals (bi-metallic) at the nanoscale offer further intriguing performance issues. The different metals may result in mixtures of metals, alloys or heterogeneous structures, each of which my exhibit different physical properties and/or performance characteristics. Applications for bi-metallic nanoparticulate metals include electronics and computing devices, bionanotechnology, medical treatment and diagnosis and energy generation and storage. The use of these bi-metallic nanometals for a variety of applications requires efficient and safe approaches for manufacturing such materials.
- In general, two fundamentally different approaches have been used to manufacture bi-metallic nanomaterials and they are referred to as “top-down” and “bottom-up” approaches. In the top-down approach, bi-metallic nanomaterials are manufactured from larger entities typically, without atomic-level control. Typcial top-down approaches include such techniques as photolithography and electron-beam lithography which start with large materials and use either machining or etching techniques to make small materials. Laser ablation is also a known top-down approach.
- In contrast, in the “bottom-up” approach, bi-metallic nanomaterials are manufactured from two or more molecular components which are caused to be assembled into bi-metallic nanoparticulate materials. In this regard, building blocks are first formed and then the building blocks are assembled into a final nano-material. In the bottom-up approach, there are a variety of general synthetic approaches that have been utilized. For example, several bi-metallic approaches include templating, chemical synthesis, sonochemical approaches, electrochemical approaches, sonoelectrochemical approaches, thermal and photochemical reduction methods including γ-ray, x-ray, laser and microwave, each of which has certain negative process and/or product limitations associated therewith.
- Whichever approach is utilized, results of bi-metallic particle size control, particle size distribution, shape control, configuration or structure control, ability to scale up, and compatability of the formed bi-metallic nanomaterial in the ultimate application, are all issues to be considered.
- In the case where two metals are formed into bi-metallic nanoparticles, further considerations such as whether the bi-metallic nanoparticles are alloys, partial alloys or partially phase segregated or completely phase segregated are also important because the specific configuration of the nanoparticles can result in different performance (e.g., biologic or catalytic). A variety of techniques exist for forming two different metals into a variety of bi-metallic nanoparticles, some of which are discussed below.
- Michael Faraday is credited with making the first colloidal gold suspension by chemical reduction methods around the 1850's (Faraday, 1857). Faraday used reduction chemistry techniques to reduce chemically an aqueous gold salt, chloroaurate (i.e., a gold (III) salt), utilizing either phosphorous dispersed into ether (e.g., CH3—CH2—O—CH2—CH3), or carbon disulfide (i.e, CS2), as the reductant.
- Today, most colloidal gold preparations are made by a reduction of chloric acid (hydrogen tetrachloroaurate) with a reductant like sodium citrate to result in “Tyndall's purple.” There are now a variety of “typical” reduction chemistry methods used to form colloidal gold. Specifically, several classes of synthesis routes exist, each of which displays different characteristics in the final products (e.g., colloidal gold nanoparticles) produced thereby. It has been noted that in addition to the strength, amount and type of the reductant utilized, the action of a stabilizer (i.e., the chemical utilized in the solution phase synthesis process) is critical (Kimling, 2006).
- While Faraday introduced colloidal gold solutions, the homogenous crystallization methods of Turkevich and Frens (and variations thereof) are most commonly used today and typically result in mostly spherical-shaped particles over a range of particle sizes (Kimling, 2006). Specifically, most current methods start with a gold (III) complex such as hydrogen tetrachloroaurate (or chloric acid) and reduce the gold in the gold complex to gold metal (i.e., gold (0) or metallic gold) by using added chemical species reductants, such as Na thiocyanate, White P, Na3 citrate & tannic acid, NaBH4, Citric Acid, Ethanol, Na ascorbate, Na3 citrate, Hexadecylaniline and others (Brown, 2008).
- Metal nanoparticle synthesis in solution(s) commonly requires the use of surface-active agents (surfactants) and/or amphiphilic polymers as stabilizing agents and/or capping agents. It is well known that surfactants and/or amphiphilic polymers serve critical roles for controlling the size, shape and stability of dispersed particles (Sakai, 2008).
- Bi-metallic nanocrystals have been formed by a number of different techniques including forming nanoparticles from the solid, gaseous and solution states. The solid state typically requires high temperature heating and annealing. The typical gaseous state approaches usually utilize molecular beam techniques, namely, the vaporization of mixed metallic powder by lasers, pulsed-arc beams, etc. However, the solution state is the much more heavily utilized bi-metallic nanoparticle formation technique. In a typical solution-based procedure, the proper chemical reactants (e.g., metal-based salts and reductants and/or stabilizers), proper control of certain intermediate reactions (which can or do occur), and control of corresponding crystallization reactions are required to achieve desired metallic nanoparticles (Wang, 2011). Further, different types of bi-metallic nanocrystals can be achieved such as a core/shell (also known as a hetero-aggregate), a hetero-structure or hetero-aggregate, an intermetallic, a mixture or alloy, as well as various core shell arrangements (Wanjala, 2011). All of these different types of bi-metallic nanocrystals can have quite different physical performance capabilities.
- In addition, it is known that making gold-platinum alloys can be quite difficult because such alloys are meta-stable and difficult to prepare (Zhou, 2007). Typical manufacturing difficulties arise from a variety of processing issues including the different oxidation-reduction potentials that exist for different metals/metal ions. Further, it is known that when platinum and gold are alloyed, the bi-metallic Pt—Au nanoparticles display unique physiochemical properties different from those of mono-metallic and non-alloyed solids (Hernandez-Fernandez, 2007).
- A variety of different approaches exist for the formation of Pt—Au bi-metallic core-shell nanostructures, but typically gold is located at the core and platinum is located on the surface of the formed bi-metallic nanocrystals. It is relatively easy to make such core-shell structures due to the different reduction potentials of typical Au ions and Pt ions in a solution (Ataee-Esfahani, 2010).
- Further, awareness is now growing that the reductant and/or stabilizers and/or other raw material components used during the formation of nanoparticles in general, including bi-metallic Pt—Au nanoparticles, may have a very large effect on the resultant performance of the nanoparticles. In particular, for example, while many have historically observed and reported on differential performance of nanoparticles due to size and shape of the nanoparticle effects (i.e., it is believed that size and shape dictate performance), only recently have attempts been made to quantify the effects of materials present at the surface of the nanoparticle. The presence of impurities such as those coming from a variety of stabilizers and/or reductants and/or the raw materials used during the manufacturing of nanoparticles, may alter performance more dramatically than size and shape alone (e.g., size and shape mabe be secondary, in some cases, to surface chemistry). In this regard, some are now “sounding an alert” that the stabilizer effect (e.g., impurities on the surface of nanoparticles) on properties of nanoparticles induces changes in their catalytic properties. Thus, consideration of how the nanoparticles were formed and their particular surface chemistry is paramount in understanding their performance characteristics (Zhang, 2010).
- Further, it has been noted that the considerable amount of surfactants and dispersants used are also a concern because such additives complicate the assessment of the true catalytic activity of a platinum surface (e.g., the performance of the nanoparticle) (Roy, 2012).
- Since the importance of nanoparticle surface chemistry is now beginning to be focused on as a key for understanding and controlling nanoparticle performance issues, attempts are now being made to remove constituents associated with manufacturing processes that are located on the surface of the formed nanoparticle (e.g., the outer layer or the presence of constituents formed as a result of reducing agent and/or surface capping agent and/or other raw materials used) including going so far as utilizing an oxygen plasma combined with electrochemical stripping (Yang, 2011). However, such surface modification approaches result in their own changes to the nanoparticle surface.
- Some have measured certain properties associated with the surface morphology (i.e., constituents located on the nanoparticle surface as a function of the formation process) and concluded that the final surface morphology of nanoparticles affects their underlying catalytic activity, perhaps even more than size and shape effects (Liang, 2007).
- In some cases, the reductant surface coating or film is permitted to remain as an impurity on the surface of the nanoparticles, but in other cases, it is attempted to be removed by a variety of somewhat complex and costly techniques. When removed, the coating typically is replaced by an alternative composition or coating to permit the nanoparticles to stay in suspension when hydrated. The influence of surface purity on the chemistry and properties of nanoparticles is often overlooked; however, results now indicate that the extent of purification can have a significant impact (Sweeney, 2006). These researchers noted that sufficient purification of nanoparticles can be more challenging that the preparation itself, usually involving tedious, time-consuming and wasteful procedures such as extensive solvent washes and fractional crystallization. Absent such purification, the variables of surface chemistry-related contaminants on the surface of chemically reduced nanoparticles affects the ability to understand/control basic structure-function relationships (Sweeney, 2006).
- Subsequent processing techniques may also require a set of washing steps, certain concentrating or centrifuging steps, and/or subsequent chemical reaction coating steps, all of which are required to achieve desirable results and certain performance characteristics (e.g., stabilization due to ligand exchange, efficacy, etc.) for the nanoparticles and nanoparticle suspensions (Sperling, 2008). In other cases, harsh stripping methods are used to ensure very clean nanoparticle surfaces (Panyala, 2009).
- Thus, others have concluded that the development of nanoparticles in the management, treatment and/or prevention of diseases is hampered by the fact that current manufacturing methods for nanoparticles are by-and-large based on chemical reduction processes. Specifically, Robyn Whyman, in 1996, recognized that one of the main hindrances in the progress of colloidal golds manufactured by a variety of reduction chemistry techniques was the lack of any “relatively simple, reproducible and generally applicable synthetic procedures” (Whyman 1996).
- Others have begun to recognize the inability to extricate completely adverse physical/biological performance of the formed nanoparticles from the chemical formation (i.e., chemical reduction) processes used to make them. In this regard, even though somewhat complex, expensive and non-environmentally friendly, washing or cleaning processes can be utilized to attempt to alter or to clean the surface of nanoparticles produced by reduction chemistry, elements of the chemical process may remain and affect the surface of nanoparticles (and thus their functioning, including biological efficacy and/or toxicity).
- Others have developed methods for removal of PVP by a facile and novel chemical method combined with minimization of chemical changes during removal (Monzo, 2012) in order to attempt to achieve clean nanoparticle surfaces. However, removal of such materials through traditional washing approaches remain elusive.
- In each of the colloidal compositions produced by reduction chemistry approaches, it is apparent that a surface coating comprising one or more elements of the reductant and/or the surfactant or capping agent will be present on (or in) at least a portion of the suspended nanoparticles. The use of a reductant (i.e., a reducing agent) may assist in suspending the nanoparticles in the liquid (e.g., water). However, the reducing agent coating or surface impurity is sometimes added to or even replaced by surfactant coatings or capping agents. Such reductant/surfactant coatings or films can be viewed as impurities located on and/or in the metal-based nanoparticles and may result in such colloids or sols actually possessing more of the properties of the protective coating or film than the nanoparticle per se (Weiser, p. 42, 1933).
- For example, surfactants and amphiphilic polymers become heavily involved not only in the formation of nanoparticles (thus affecting size and shape), but also in the nanoparticles per se. Surface properties of the nanoparticles are modified by reductant coatings and/or surfactant molecule coatings (Sperling, 2008).
- A variety of sonoelectrochemical techniques exist for producing both single metallic nanoparticles and bi-metallic nanoparticles. Sonoelectrical processes typically direct electric and acoustic energy toward metal-based raw material salts (e.g., HAuCl4·4H2O (AuCl4 −), NaAuCl4·2H2O, H2PtCl6·6H2O, HAuCl3·3H2O, etc.) and metal ions in those salts are caused to be reduced by one or more reductant species created by the sonoelectrochemial method. In this regard, often a single electrode induces the growth of nanoparticles thereon by an electrochemical step, followed by an acoustic step which, more or less, attempts to eject the nanoparticles off from the electrode and also creates additional reductant material by, for example, lysis of water molecules. In this regard, a single electrode typically performs a dual duty of both electrochemistry (e.g., nanoparticle formation) and acoustic chemistry (e.g., reductant formation) (Nagata, 1996).
- Most of the sonoelectrochemical techniques utilize one or more reductants and/or capping agents in addition to any of those which may be formed in situ by the process. In this regard, a variety of different polymers have been utilized as capping agents for single metallic nanoparticles (Saez, 2009). However, work by others (Liu, 2004; Ou, 2011; Mai, 2011; and Liu, 2006) all disclose similar sonoelectrochemical techniques for making gold nanoparticles with sonoelectrochemical pulse methods using, allegedly, no added reductants. For example, utilization of an acid solution in combination with electrochemical cycling to strip gold ions from a gold electrode and form AuCl4 − compounds in an aqueous solution has been disclosed (Liu, 2004). Subsequently, the gold ions are reduced by created reductant species (e.g., lysis products of H2O) produced in their sonoelectrochemical process. Apparently, however, the concentrations of gold nanoparticles produced are quite limited by this technique (e.g., 3 ppm) without the addition of other materials (e.g., stabilizers) (Ou, 2011).
- Alternative sonoelectrochemical methods have been used to make gold nanoparticles. Specifically, starting materials of HAuCl4·4H2O and KNO3 were pH-adjusted by adding NaOH to obtain different pH's, with a pH of of about 10 being noted as optimal. Nanoparticles having diameters of approximately 20 nm were produced. The surface potential of the gold nanoparticles around the pH of 10 was −54.65 mV. It was concluded that the OH− groups adsorbed on gold nanoparticles and caused electrostatic repulsion therebetween. Thus, no added reductants were necessary (Shen, 2010).
- A variety of sonoelectrochemical technquies have also been set forth for making bi-metallic nanoparticles. For example, platinum-gold nanoparticles stabilized by PEG-MS (polyetholeneglycolmonostearate) have been manufactured (Fujimoto, 2001). Further, binary gold/platinum nanoparticles made by sonoelectrochemistry utilizing surfactants (anionic surfactants; sodium dodechal sulfate (SDS) or nonionic surfactant polyetholeneglycolmonostearate PEG-MS) have also been made (Nakanishi, 2005). In this method, the addition of some surfactants is reported as being indispensable (Nakanishi, 2005). Likewise, in some related work, the use of SDS or PEG-MS in combination with various sonoelectrochemical techniques has been reported (Takatani, 2003). These bi-metallic nanocrystals made by sonoelectrochemical techniques all require the use of surfactants.
- Radiolytic techniques for making nanoparticles have been directed primarily to single-metals (i.e., not bi-metals). Another older and more complex technique for minimizing or eliminating the need for reducing agents and/or minimizing undesirable oxidation products of the reductant utilizes γ-irradiation from a 60Co source at a dose rate of 1.8×104 rad/h. In this instance, Au(CN)2 was reduced by first creating hydrated electrons from the radiolysis of water and utilizing the hydrated electrons to reduce the gold ions, namely:
-
e aq −+Au(CN)2→Au0+2CN− (Henglein, 1998). - Futher, the creation of hydrated electrons and OH radicals by pulse activation from a linear accelerator has also occurred (Ghosh-Mazumdar, 1968). Such created species assist in the reduction of various metals from aqueous metallic-based salts.
- Most work using x-rays for the manufacture of metal-based nanoparticles has been focused on single metal composition metallic-based nanoparticles, however, some recent work on intense x-ray radiation has also occurred to make alloys (with surfactants).
- The use of synchrotron x-ray synthesis of HAuCl4, with added NaCO3, has been used to make colloidal gold nanoparticles without adding additional reducing agent (Yang, 2006). In this technique, a gold salt was dissolved to make a solution and an appropriate amount of NaHCO3 was added thereto. The reported result was particle sizes of 10-15 nm, as measured, a pH of about 7 and the gold suspensions were relatively stable due to the coordination of OH− groups around the gold nanoparticles (Yang, 2006).
- Single metal gold nanosols stabilized by electrostatic protection due to x-ray irradiation has also occurred (Wang, 2007; Wang, 2007). The x-rays generated reductant electrons in the precursor solutions. It was noted that this approach required very intense x-ray beams (thus requiring synchrotron sources) (Wang, 2007; Wang, 2007). Additionally, the nanoparticle suspensions were formed with a pH of 9 and had a surface potential of −57.8+/−mV, as measured by a zeta meter. The formed nanoparticles were about 10 nm in size. Additionally, modification of the pH to values between 6-9 occurred by adding NaOH to the solution (Wang, 2007). Further, the x-rays used are well above the threshold energy for water radiolysis and additional x-ray energy may be causing intermediate reactions that they do not recognize (e.g., kinetic effects) (Wang, 2007).
- Further, x-ray photochemical reactions have been used to make gold nanoparticle suspensions (Ma, 2008). It was noted that knowledge of the details of the intermediate reactions prior to nanoparticle formation is critical to controlling size, shape and properties (Ma, 2008).
- A one-pot synthesis of Au—Pt alloys by intense x-ray irradiation has also been disclosed (Wang, 2011). The incident x-rays irradiate a gold/platinum salt solution (i.e., HAuCl4·3H2O and H2PtCl6.6H2O) containing PEG (a common surfactant molecule known to prevent nanoparticle aggregation). However, it was noted that PEG could negatively impact applications that are sensitive to surface conditions, such as catalysis (Wang, 2011).
- Bi-metallic Pt—Au nanoparticles have been made by femtosecond laser synthesis (Chau, 2011). Specifically, gold and platinum salt solutions (i.e., HAuCl4·4H2O, H2PtCl6.6H2O) were combined with PVP (a known dispersing/stabilizing agent) and the solution was laser irradiated. In related work, high intensity laser radiation of a similar solution of gold and platinum salts occurred. However, in this solution no PEG was added and the resultant nanoparticles were found not to be stable (Nakamura, 2011; Nakamura, 2010; Nakamura, 2009).
- A top-down laser ablation approach to make gold nanoparticles has also been attempted. However, laser ablation typically results in some sort of oxide on the surface of the metal target (Sylvestre, 2004).
- Bi-metallic gold-platinum nanoparticles have also been made by electron beam irradiation (Mirdamadi-Esfahani, 2010). Specifically, in this approach, the electron beam irradiation creates hydrated electrons and reducing radicals due to the radiolysis of water. Metal salts of gold and platinum (i.e., KAuCl4 and H2PtCl6) are mixed with polyacrylic acid (i.e., a dispersant/stabilizing agent) and accelerated electrons are directed thereto.
- Different surface chemistries or surface films (e.g., the presence of reductant by-product compositions and/or thicknesses (e.g., films) of reductants or reductant by-products) can result in different interactions of the nanoparticles with, for example, a variety of proteins in an organism. Biophysical binding forces (e.g., electrostatic, hydrophobic, hydrogen binding, van der Waals) of nanoparticles to proteins are a function not only of the size, shape and composition of the nanoparticles, but also the type of and/or thickness of the surface impurities or coating(s) on the nanoparticles (Lacerda, 2010).
- A better understanding of the biological effects of nanoparticles requires an understanding of the binding properties of the in-vivo proteins that associate themselves with the nanoparticles. Protein absorption (or a protein corona) on nanoparticles can change as a function of nanoparticle size and surface layer composition and thickness. Protein layers that “dress” the nanoparticle control the propensity of the nanoparticles to aggregate and strongly influence their interaction with biological materials (Lacerda, 2010).
- Additionally, both the shape and the surface chemistry of nanoparticles influenced cytotoxicity and cellular uptake in model biological systems (Qiu, 2010). However, it was concluded that only the surface chemistry contributes to undesirable cytotoxicity. In particular, it was shown that CTAB-coated (i.e., cetyltrimethlammonium bromide) gold nanoparticles release portions of their coatings at different points in a biological process and/or different location(s) within an organism, which results in toxicity (Qui, 2010).
- Further, in an important article published in 2010, the authors state that since 1981, more than 230 published studies utilize gold nanoparticles generated from the citrate reduction method with scarce data on non-gold components in the reaction system (Balassubramanian, 2010). The authors conclude it is clear that much of the testing of biological performance has been skewed by the lack of understanding of components present in/on the nanoparticles (e.g., the surface chemistry) other than nanoparticles per se (Balassubramanian, 2010).
- The protein corona which forms on a nanoparticle is important because it is the protein corona that gives the biological identity to the nanoparticle (Lynch, 2007). The surface of the nanoparticle assists in the formation of the protein corona as well as its size and its shape (Lynch, 2007).
- Further, albumin-based drug delivery has been recognized as a novel therapeutic approach (Wunder, 2003; Stehle, 1997; Stehle, 1997). Specifically, the albumin-binding assists in delivery of the therapeutic to desirable targeted locations resulting in higher efficacy/lower toxicity.
- The references cited throughout the “Background of the Invention” are listed below in detail.
- Ataee-Esfahani, H., Wang, L., Nemoto, Y. & Yamauchi, Y. (2010). Synthesis of Bimetallic Au@Pt Nanoparticles with Au Core and Nanostructured Pt Shell toward Highly Active Electrocatalysts. Chem. Mater., 22, 6310-6318.
- Balasubramanian, S. K., Yang, L., Yung, L.-Y. L., Ong, C.-N., Ong, W.-Y. & Yu, L. E. (2010). Characterization, purification, and stability of gold nanoparticles. Biomaterials, 31, 9023-9030.
- Brown, C. L., Whitehouse, M. W., Tiekink, E. R. T., & Bushell G. R. (2008). Colloidal metallic gold is not bio-inert. Inflammopharmacology, 16, 133-137.
- Chau, J. L. H., Chen, C.-Y., Yang, M.-C., Lin, K.-L., Sato, S., Nakamura, T., Yang, C.-C. & Cheng, C.-W. (2011). Femtosecond laser synthesis of bimetallic Pt—Au nanoparticles. Materials Letters, 65, 804-807.
- Faraday, M. (1857). The Bakerian lecture: Experimental relations of gold (and other metals) to light. Philosoph. Trans. R. Soc. London, 147, 145-181.
- Fujimoto, T., Mizukoshi, Y., Nagata, Y., Maeda, Y. & Oshima, R. (2001). Sonolytical Preparation of Various Types of Metal Nanoparticles in Aqueous Solution. Scipta mater., 44, 2183-2186.
- Ghosh-Mazumdar, A. S. (1968). A Pulse Radiolysis Study of Bivalent and Zerovalent Gold in Aqueous Solutions. Radiation Chemistry, 193-209.
- Henglein, A. & Meisel, D. (1998). Radiolytic Control of the Size of Colloidal Gold Nanoparticles. Langmuir, 14, 7392-7396.
- Hernandez-Fernandez, P., Rojas, S., Ocon, P., Gomez de la Fuente, J. L., San Fabian, J., Sanza, J., Pena, M. A., Garcia-Garcia, F. J., Terreros, P. & Fierro, J. L. G. (2007) Influence of the Preparation Route of Bimetallic Pt—Au Nanoparticle Electrocatalysts for the Oxygen Reduction Reaction. J. Phys. Chem. C, 111, 2913-2923.
- Kimling, J., Maier, M., Okenve, B., Kotaidis, V., Ballot, H. & Plech, A. (2006). Turkevich Method for Gold Nanoparticle Synthesis Revisited. J. Phys. Chem. B, 110, 15700-15707.
- Lacerda, S. H. D. P., et al. (2010). Interaction of Gold Nanoparticles with Common Human Blood Proteins. American Chemical Society, 4 (1), 365-379.
- Liu, Y.-C., Lin, L.-H. & Chiu, W.-H. (2004) Size-Controlled Synthesis of Gold Nanoparticles from Bulk Gold Substrates by Sonoelectrochemical Methods. J. Phys. Chem. B, 108, 19237-19240.
- Liu, Y.-C., Yu, C.-C, & Yang, K.-H. (2006). Active catalysts of electrochemically prepared gold nanoparticles for the decomposition of aldehyde in alcohol solutions. Electrochemistry Communications, 8, 1163-1167.
- Liu, Y.-C., Lee, H.-T. & Peng, H.-H. (2004). New pathway for sonoelectrochemical synthesis of gold-silver alloy nanoparticles from their bulk substrates. Chemical Physics Letters, 400, 436-440.
- Lynch, I., Cedervall, T., Lundqvist, M., Cabaleiro-Lago, C., Linse, S. & Dawson, K. A. (2007). The nanoparticle-protein complex as a biological entity; a complex fluids and surface science challenge for the 21st century. Advances in Colloid and Interface Science, 134-135.
- Ma, Q., Divan, R., Mancini, D. C. & Keane, D. T. (2008). Elucidating Chemical and Morphological Changes in Tetrachloroauric Solutions Induced by X-ray Photochemical Reaction. J. Phys. Chem. A., 112, 4568-4572.
- Mai, F.-D., Hsu, T.-C., Liu, Y.-C. & Yang, K.-H. (2011). Strategy of fabricating enriched gold nanoparticles based on electrochemical methods. Materials Letters, 65, 1788-1790.
- Mirdamadi-Esfahani, M., Mostafavi, M., Keita, B., Nadjo, L., Kooyman, P. & Remita, H. (2010). Bimetallic Au—Pt nanoparticles synthesized by radiolysis: Application in electro-catalysis. Gold Bulletin, 43, (1).
- Monzo, J., Koper, M. T. M. & Rodriguez, P. (2012). Removing Polyvinylpyrrolidone from Catalytic Pt Nanoparticles without Modification of Superficial Order. ChemPhysChem, 13, 709-715.
- Nagata, Y., Mizukoshi, Y. Okitsu, K. & Maeda, Y. (1996). Sonochemical Formation of Gold Particles in Aqueous Solution. Radiation Research, 146, 333-338.
- Nakamura, T., Herbani, Y. & Sato, S. (2011). Fabrication of Gold-Platinum Nanoalloy by High-Intensity Laser Irradiation of Solution. Supplemental Proceedings, 2, 3-8.
- Nakamura, T., Herbani, Y. & Sato, S. (2010). Fabrication of gold-platinum nanoparticles by intense, femtosecond laser irradiation of aqueous solution. Optical Society of America.
- Nakamura, T., Magara, H., Herbani, Y., Ito, A. & Sato, S. (2009). Fabrication of gold-platinum nanoparticles by intense, femtosecond laser irradiation of aqueous solution. Optical Society of America.
- Nakanishi, M., Takatani, H., Kobayashi, Y., Hori, F., Taniguchi, R., Iwase, A. & Oshima, R. (2005). Characterization of binary gold/platinum nanoparticles prepared by sonochemistry technique. Applied Surface Science, 241, 209-212.
- Ou, K.-L., Yang, K.-H., Liu, Y.-C., Hsu, T.-C. & Chen, Q.-Y. (2011). New strategy to prepare enriched and small gold nanoparticles by sonoelectrochemical pulse methods. Electrochimica Acta, 58, 497-502.
- Panyala, N. G., Pena-Mendez, E. M., & Havel, J. (2009). Gold and nano-gold in medicine: overview, toxicology and perspectives. Journal of Applied Biomedicine, 7, 75-91.
- Qiu, Y., et al. (2010). Surface chemistry and aspect ratio mediated cellular uptake of Au nanorods. Biomaterials, 31, 7606-7619.
- Roy, R. K., Njagi, J. I., Farrell, B., Halaciuga, I., Lopez, M. & Goia, D. V. (2012). Deposition of continuous platinum shells on gold nanoparticles by chemical precipitaion. Journal of Colloid and Interface Science, 369, 91-95.
- Saez, V. & Mason, T. J. (2009). Sonoelectrochemical Synthesis of Nanoparticles. Molecules, 14, 4284-4299.
- Sakai, T., Enomoto, H., Torigoe, K., Kakai, H. & Abe, M. (2008). Surfactant- and reducer-free synthesis of gold nanoparticles in aqueous solutions. Colloids and Surface A: Physiocochemical and Engineering Aspects, 18-26.
- Shen, Q., Min, Q., Shi, J., Jiang, L., Hou, W. & Zhu, J.-J. (2011). Synthesis of stabilizer-free gold nanoparticles by puls sonoelectrochemical method. Ultrasonics Sonochemistry, 18, 231-237.
- Sperling, R. A., Gil, P. R., Zhang, F., Zanella, M., & Parak, W. J. (2008). Biological applications of gold nanoparticles. Chem. Soc. Rev, 37, 1896-1908.
- Stehle, G., Sinn, H., Wunder, A., Schrenk, H. H., Schutt, S., Maier-Borst, W. & Heene, L. (1997). The loading rate determines tumor targeting properties of methotrexate-albumin conjugates in rats. Anti-Cancer Drugs, 8, 677-685.
- Stehle, G., Wunder, A., Sinn, H., Schrenk, H. H., Schutt, S., Frei, E., Hartung, G., Maier-Borst, W. & Heene, D. L. (1997). Pharmacokinetics of methotrexate-albumin conjugates in tumor-bearing rats. Anti-Cancer Drugs, 8, 835-844.
- Sweeney, S. F., Woehrle, G. H. & Hutchison, J. E. (2006). Rapid Purification and Size Separation of Gold Nanoparticles via Diafiltration. J. Am. Chem. Soc., 128, 3190-3197.
- Sylvestre, J.-P., Poulin, S., Kabashin, A. V., Sacher, E., Meunier, M. & Luong, J. H. T. (2004). Surface Chemistry of Gold Nanoparticles Produced by Laser Ablation in Aqueous Media. J. Phys. Chem. B., 16864-16869.
- Takatani, H., Kago, H., Nakanishi, M., Kobayashi, Y., Hori, F. & Oshima, R. (2003). Characterization of Noble Metal Alloy Nanoparticles Prepared by Ultrasound Irradiation. Rev. Adv. Mater. Sci., 5, 232-238.
- Wang, C. H., et al. (2007). Aqueous gold nanosols stabilized by electrostatic protection generated by X-ray irradiation assisted radical reduction. Materials Chemistry and Physics, 106, 323-329.
- Wang, D. & Li, Y. (2011). Bimetallic Nanocrystals: Liquid-Phase Synthesis and Catalytic Applications. Adv. Mater, 23, 1044-1060.
- Wang, C.-H., Hua, T.-E., Chien, C.-C., Yu, Y.-L., Yang, T.-Y., Liu, C.-J., Leng, W.-H., Hwu, Y., Yang, Y.-C., Kim, C.-C., Je, J.-H., Chen, C.-H., Lin, H.-M. & Margaritondo, G. (2007). Aqueous gold nanosols stabilized by electrostatic protection generated by X-ray irradiation assisted radical reduction. Materials Chemistry and Physics, 106, 323-329.
- Wang, C.-H., Chien, C.-C., Yu, Y.-Lu., Liu, C.-J., Lee, C.-F., Chen, C.-H., Hwu, Y., Yang, C.-S., Je, J.-H. & Margaritondo, G. (2007). Structural properties of ‘naked’ gold nanoparticles formed by synchrotron X-ray irradiation. J. Synchrotron Rad., 14, 477-482.
- Wang, C.-L., Hsao, B.-J., Lai, S.-F., Chen, W.-C., Chen, H.-H., Chen, Y.-Y., Chien, C.-Ch. Cai, X., Kempson, I. M., Hwu, Y. & Margaritondo, G. (2011). One-pot synthesis of AuPt alloyed nanoparticles by intense x-ray irradiation. Nanotechnology, 22, 065605-065611.
- Wanjala, B. N., Luo, J., Fang, B., Mott, D. & Zhong, C-J. (2011). Gold-platinum nanoparticles: alloying and phase segregation. J. Mater. Chem, 21, 4012-4020.
- Weiser, H. B. Inorganic Colloid Chemistry—Volume I: The Colloidal Elements. New York: John Wiley & Sons, Inc., 1933.
- Whyman, R. (1996). Gold Nanoparticles A Renaissance in Gold Chemistry. Gold Bulletin, 29(1), 11-15.
- Wunder, A. Muller-Ladner, U., Stelzer, E. H. K., Funk, J., Neumann, E., Stehle, G., Pap, T., Sinn, H., Gay, S. & Fiehn, C. (2003). Albumin-Based Drug Delivery as Novel Therapeutic Approach for Rheumatoid Arthritis. The Journal of Immunology, 170, 4793-4801.
- Yang, S., Park, N.-Y., Han, J. W., Kim, C., Lee, S.-C. & Lee, H. (2012). A distinct platinum growth mode on shaped gold nanocrystals. Chem. Commun, 48, 257-259.
- Yang, Y.-C., Wang, C.-H., Hwu, Y.-K. & Je, J.-H. (2006). Synchrotron X-ray synthesis of colloidal gold particles for drug delivery. Materials Chemistry and Physics, 100, 72-76.
- Zhang, G.-R. & Xu, B.-Q. (2010). Surprisingly strong effect of stabilizer on the properties of Au nanoparticles and Pt{circumflex over ( )}Au nanostructures in electrocatalysts. Nanoscale, 2, 2798-2804.
- Zhou, S., Jackson, G. S. & Eichhorn, B. (2007). AuPt Alloy Nanoparticles for CO-Tolerant Hydrogen Activation: Architectural Effects in Au—Pt Bimetallic Nanocrystals. Adv. Funct. Mater., 17, 3099-3104.
- New bi-metallic nanocrystal suspensions are provided that have nanocrystalline surfaces that can be substantially free (as defined herein) from organic or other impurities or films, or in certain cases may contain some desirable film or partial coating. Specifically, the surfaces are “clean” relative to those made using chemical reduction processes that require chemical reductants and/or surfactants to grow gold nanoparticles from metal ions in solution. Resulting bi-metallic nanocrystalline suspensions or colloids have desirable pH ranges such as 4.0-12.0, but more typically 5.0-11.0, and even more typically 8.0-11.0, and in many embodiment, 10.0-11.0 and zeta potential values of at least −20 mV, and more typically at least −40 mV, and even more typically at least −50 mV for the pH ranges of interest.
- The shapes and shape distributions of these bi-metallic nanocrystals prepared according to the manufacturing process described below include, but are not limited to, spheres, pentagons, hexagons (e.g., hexagonal bipyramids, icosahedrons, octahedrons), and “others”.
- Any desired average size of bi-metallic nanocrystals below 100 nm can be provided. The most desirable crystalline size ranges include those having an average crystal size (as measured and determined by specific techniques disclosed in detail herein) that is predominantly less than 100 nm, and more typically less than 50 nm, even more typically less than 30 nm, and in many of the preferred embodiments disclosed herein, the average crystal size for the nanocrystal size distribution is less than 20 nm and with an even more preferable range of 8-18 nm. However, for certain applications, the electrochemical techniques disclosed herein can be utilized to result in larger nanocrystals, if desired.
- A variety of concentrations of bi-metallic nanocrystals can be provided according to the invention. For example, total atomic metal concentrations of bi-metallic nanocrystals produced initially can be a few parts per million (i.e., μg/ml or mg/l) up to a few hundred ppm, but are typically in the range of 2-200 ppm (i.e., 2 μg/ml-200 μg/ml) and more often in the range of 2-50 ppm (i.e., 2 μg/ml-50 μg/ml) and even more typically 5-20 ppm (i.e., 5 μg/ml-20 μg/ml). However, novel concentration techniques are disclosed herein which allow concentrated “initial” product to be formed with ppm's between 200-5,000 ppm and more preferably, 200-3,000 ppm and more preferably, 200-1,000 ppm.
- The bi-metallic nanocrystals in suspension can be made as alloys, partial alloys, phase-segregated or heteroaggregates or mixtures. In preferred embodiments herein, the bi-metallic nanocrystals are alloys and/or heteroaggregates. Gold is typically the major constituent (i.e., more by weight and more by volume) and platinum is typically the minor constituent (i.e., less by weight and less by volume). Typical ratios range from 2/1 to 10/1, with preferred ranges being 3/1 to 8/1, and even more preferred 3/1 to 6/1.
- A novel set of processes are provided to produce these unique bi-metallic nanocrystals. Each process involves the creation of the bi-metallic nanocrystals in water. In a preferred embodiment, the water contains an added “process enhancer” which does not significantly bind to the formed nanocrystals, but rather facilitates nucleation/crystal growth during the electrochemical-stimulated growth process. The process enhancer serves important roles in the process including, for example, providing charged ions in the electrochemical solution to permit the crystals to be grown.
- In a preferred embodiment, a first step includes forming a platinum metal-based species with at least one process enhancer and the formed aqueous suspension/solution is then used as a raw material solution/suspension in a second step where a gold metal-based species is reduced and/or co-reduced to grow the bi-metallic nanocrystals in water. Specifically, the processes involve first forming electrochemically at least one platinum species in water and at least one lysis product of water, thereby creating a platinum species and water material; and using the created platinum/water material in a second electrochemical reaction to form a suspension of bi-metallic gold-platinum nanocrystals in water.
- By following the inventive electrochemical manufacturing processes of the invention, these bi-metallic nanocrystals can form alloys or metal “coatings” (or portions of coatings, e.g., islands) on core metals or alternatively, form heteroaggregates. Alternatively, a mixture of nanocrystals can be made. Also, a range of alloys or mixtures or heteroaggregates may result within a single colloid or suspension, if desired. In some cases, desirable residual metal ions may be in solution in the suspension.
- These novel electrochemical processes can occur in either a batch, semi-continuous or continuous process. These processes result in controlled bi-metallic nanocrystalline concentrations, controlled nanocrystal sizes and controlled nanocrystal size ranges. Novel manufacturing assemblies are provided to produce these bi-metallic nanocrystals.
- Since these bi-metallic nanocrystals have substantially cleaner surfaces than the prior available metallic-based (or bi-metallic-based) nanoparticles, and can desirably contain spatially extended low index crystallographic planes forming novel crystal shapes and/or crystal shape distributions, the bi-metallic nanocrystals appear to be more active (e.g., more biologically active and may be less toxic) relative to those containing surface contaminants such as chemical reductants and/or surfactants or residual raw materials that result from traditional chemical reduction (or other) processes. Therefore, uses for nanoparticles, such as, catalysis processes, medical treatments, biologic processes, medical diagnostics, etc., may be affected at lower concetrations of metallic-based nanocrystals made according to the techniques herein.
- Further, because the raw material metal ions used to grow the bi-metallic nanocrystals are provided by sacrificial metal electrodes used during the various electrochemical processes, there are no requirements for gold-based salts (or the equivalent) or platinum-based salts (or the equivalent) to be provided as raw materials for the formation of Au—Pt bi-metallic nanocrystal suspensions. Accordingly, components such as Cl−, chlorides or chlorine-based materials are not required to be part of the novel process or part of the novel bi-metallic nanocrystal suspensions produced. Additionally, no chlorine-based acids are required to produce the Au—Pt bi-metallic suspensions.
- Still further, the aforementioned metal-based bi-metallic nanocrystal suspensions or colloids of the present invention can be mixed or combined with other metallic-based solutions or colloids to form novel solution or colloid mixtures (e.g., in this instance, distinct metal species can still be discerned, etiher as composites or distinct species in a suspension).
-
FIG. 1 shows a schematic cross-sectional view of a manual electrode assembly according to the present invention. -
FIG. 2 shows a schematic cross-sectional view of an automatic electrode control assembly according to the present invention. -
FIGS. 3 a-3 e show five different representative embodiments of configurations for theelectrode 1. -
FIG. 4 shows a cross-sectional schematic view of plasmas produced utilizing one specific configuration of theelectrode 1 corresponding toFIG. 3 e. -
FIGS. 5 a-5 e show a variety of cross-sectional views ofvarious trough members 30. -
FIG. 6 shows a schematic cross-sectional view of a set ofcontrol devices 20 located on atrough member 30 with aliquid 3 flowing therethrough and into astorage container 41. -
FIG. 7 a shows an AC transformer electrical wiring diagram for use with different embodiments of the invention. -
FIG. 7 b shows a schematic view of atransformer 60 andFIGS. 7 c and 7 d show schematic representations of two sine waves in phase and out of phase, respectively. -
FIG. 8 a shows a view ofgold wires -
FIG. 8 b shows a view of thegold wires -
FIG. 8 c shows thedevice 20 used in all trough Examples herein that utilize a plasma. -
FIGS. 8 d, 8 e, 8 f and 8 g show wiring diagrams used to monitor and/or control thedevices 20. -
FIGS. 8 h and 8 i show wiring diagrams used topower devices 20. -
FIG. 8 j shows a design for poweringwires 5/5 in thedevices 20. -
FIG. 9 shows afirst trough member 30 a′ wherein oneplasma 4 a is created. The output of thisfirst trough member 30 a′ flows into asecond trough member 30 b′. -
FIGS. 10 a-10 d show an alternative design of thetrough member 30 b′ wherein thetrough member portions 30 a′ and 30 b′ are contiguous. -
FIGS. 11 a-11 b show twotrough members 30 b′ used in connection withFIGS. 10 a-10 d and various Examples herein. -
FIG. 11 c shows a representative TEM photomicrograph of dried gold constituents formed in connection with Example 1. -
FIG. 11 d shows a particle size distribution histogram from TEM measurements for the constituents formed in connection with Example 1. -
FIG. 11 e shows the UV-Vis spectral patterns of each of the gold suspension made according to Example 1. -
FIG. 12 a shows a schematic of an apparatus used in a batch method whereby in a first step, aplasma 4 is created to condition afluid 3′. -
FIGS. 12 b and 12 c show a schematic of an apparatus used in a batchmethod utilizing wires FIG. 12 a and as discussed in various Examples herein. -
FIG. 12 d shows a schematic of an apparatus used in a batchmethod utilizing wires FIG. 12 a , and as discussed in various examples herein. -
FIG. 12 e shows a schematic view of the amplifier used in Examples 2 and 3. -
FIG. 12 f shows a schematic view of the power supply used in Examples 2 and 3. -
FIG. 12 g shows the UV-Vis spectral pattern of the Au—Pt bi-metallic suspensions made according to Example 6. -
FIG. 13 is a schematic of the power supply electrical setup used to generate the nanocrystals in the many Examples herein. -
FIG. 14 shows a representative TEM photomicrograph of dried platinum constituents formed in connection with Example 2. -
FIG. 15 a shows a representative TEM photomicrograph of dried platinum constituents formed in connection with Example 3. -
FIG. 15 b shows a particle size distribution histogram from TEM measurements for the constituents formed in connection with Example 3. -
FIG. 16 shows a representative TEM photomicrograph of dried platinum constituents formed in connection with Example 4. -
FIG. 17 shows the UV-Vis spectral patterns of each of the seven platinum solutions/suspensions made according to Example 5. -
FIG. 18 shows a representative TEM photomicrograph of the dried constituents made according to Example 6. -
FIG. 19 shows a representative TEM photomicrograph of the dried constituents made according to Example 7. -
FIG. 20 shows a representative TEM photomicrograph of the dried constituents made according to Example 8. -
FIGS. 21 a and 21 b show representative TEM photomicrographs of dried constituents made according to Example 9. -
FIGS. 22 a and 22 b are representative EDS spectra corresponding toFIGS. 21 a and 21 b , respectively. -
FIGS. 23 a and 23 b show representative TEM photomicrographs of dried constituents made according to Example 9. -
FIGS. 24 a and 24 b are representative EDS spectra corresponding toFIGS. 23 a and 23 b , respectively. -
FIG. 25 a shows a representative TEM photomicrograph of dried constituents made according to Example 10; andFIG. 25 b is a representative EDS spectra corresponding toFIG. 25 a. -
FIG. 26 a shows a representative TEM photomicrograph of dried constituents made according to Example 11; andFIG. 26 b is a representative EDS spectra corresponding toFIG. 26 a. -
FIG. 27 shows a UV-Vis spectrograph of GPB-032. -
FIG. 28 a shows three UV-Vis spectrographs of three Au—Pt bi-metallic suspensions. -
FIG. 28 b shows UV-Vis spectrographs for five different GPB bi-metallic suspensions. -
FIG. 28 c shows a graph of particle radius versus frequency for bi-metallic nanoparticles made according to Example 16. -
FIG. 29 a shows a representative TEM photomicrograph of the dried constituents made according to Example 17. -
FIG. 29 b is a representative EDS spectra corresponding toFIG. 29 a. -
FIG. 29 c shows a representative TEM photomicrograph of the dried constituents made according to Example 17. -
FIG. 29 d is a representative EDS spectra corresponding toFIG. 29 c. -
FIGS. 29 e, 29 f and 29 g are Scanning Transmission Electron Microscopy images of nanocrystals in a GPB-040 suspension. -
FIGS. 29 h and 29 i are representative XPS spectra corresponding to Example 17. -
FIG. 30 is a UV-Vis spectrograph of GPB-040 made according to Example 17. -
FIGS. 31 a and 31 b are schematic representations of the dialysis procedure used in Example 18; andFIG. 31 c is a schematic representation of a TFF apparatus. -
FIGS. 32 a -32 ad are graphical depictions of anti-cancer activity of two suspensions (NE10214 and a bi-metallic nanocrystal suspension, GPB-032). -
FIGS. 33 a and 33 b show the results of the cancer xenograft tests set forth in Example 20a. -
FIGS. 34 a and 34 b show the results of the cancer xenograft tests set forth in Example 20b. -
FIGS. 35 a and 35 b show the results of the cancer xenograft tests set forth in Example 20c. -
FIGS. 36 a and 36 b show the results of the cancer xenograft tests set forth in Example 20d. -
FIGS. 37 a and 37 b show the results of the cancer xenograft tests set forth in Example 20e. -
FIGS. 38 a and 38 b show the results of the cancer xenograft tests set forth in Example 20f. -
FIGS. 39 a and 39 b represent the liquid consumption amount and weight gain for the mice set forth in Example 21. -
FIGS. 40 a and 40 b are graphs depicting the amount of absorbance of GPB-11 and various protein binders. -
FIG. 40 c shows an AFS photomicrograph of DNA binding to nanocrystals of GPB-11. - New aqueous-based bi-metallic nanocrystal suspensions are manufactured from a combination of gold and platinum donor electrode materials, such bi-metallic nanocrystals including nanocrystalline surfaces that can be substantially free from organic or other impurities or films. Specifically, the surfaces of the bi-metallic nanocrystals are “clean” relative to those surfaces of similar chemical composition nanoparticles made using: (1) chemical reduction processes that require chemical reductants and/or surfactants and/or various salt compounds as parts of the raw materials used to form bi-metallic-based nanoparticles from transition metal ions contained in raw material solution; and (2) other processes (including, sonoelectrochemistry, gamma-ray radiation, x-ray radiation, laser irradiation, electron accelorators, etc.) which use, for example, a variety of reductants or chlorine-based (or salt-based) raw materials (e.g., metal salts).
- The new bi-metallic nanocrystals of gold and platinum are produced via novel electrochemical manufacturing procedures, described in detail herein. The new electrochemical manufacturing procedures do not require the addition of chemical reductants and/or surfactants (e.g., organic compounds) or other agents, to be added to reduce metal ions and/or stabilize the formed bi-metallic nanocrystals. Further, the processes do not require the addition of raw materials which contain both metal ions (which are reduced to form metal nanoparticles) and associated ions or species which counterbalance the electrical charge of the positively charged metal ion(s). Such added reductants, stabilizers and non-metal ion portions of raw materials are undesirable when they are typically carried along in, or on, the particles, or are undesirably adhered to at least a portion of the surface of the chemically reduced particles and/or remain as ions in the suspension. It is now understood that certain nanocrystal performance requirements can not be met with such impurities located on or bonded to the surface and such impurities need to be subsequently stripped or removed using various undesirable processes, which process themselves can affect the surface of the nanoparticles (e.g., plasma etching).
- In a preferred embodiment, a first set of electrochemical steps of the process involves the in situ creation of platinum species (e.g., raw materials) from a platinum metal source. The platinum species is created in water which contains a “process enhancer” or “processing enhancer” (typically an inorganic material or carbonate or such) which does not significantly bind to the formed nanocrystals in suspension, but rather facilitates removal of metal ions from a donor platinum metal electrode source, and/or assists in nucleation/growth during electrochemical-stimulated nanocrystal growth processes. More specifically, the process enhancer serves important roles in the process including providing charged ions in the electrochemical solution to permit metal ions to be in solution and/or to cause the nanocrystals to be grown. The process enhancer is critically a compound(s) which remains in solution, and/or does not form a coating (e.g., an organic coating), and/or does not adversely affect the performance of the formed nanocrystals or the formed suspension(s) (e.g., is inert), and/or can be destroyed, evaporated, removed or otherwise lost during one or more steps of the electrochemical process. A preferred process enhancer is sodium bicarbonate. Examples of other process enhancers are sodium carbonate, sodium hydroxide, potassium bicarbonate, potassium carbonate, potassium hydroxide, trisodium phosphate, disodium phosphate, monosodium phosphate, potassium phosphates or the like and combinations thereof. Another particularly preferred processing enhancer is a mixture of sodium bicarbonate and potassium hydroxide.
- Desirable concentration ranges for the processing enhancer in the first step of the process include typically 0.01-20 grams/gallon (0.0026-2.1730 mg/ml), more typically, 0.1-7.5 grams/gallon (0.0264-1.9813 mg/ml) and most typically, 0.5-2.0 grams/gallon (0.13210-0.5283 mg/ml).
- Further, desirable concentrations of the platinum species made in the first electrochemical steps of the process range from about 0.5 ppm to about 20 ppm and most typically about 1-8 ppm, and even more typically about 0.5-4 ppm. The result of the first set of electrochemical steps is a platform species in water. The platinum species can be predominantly nanocrystals or a mixture of nanocrystals and platinum ions. In a preferred embodiment, the platinum species is predominantly ions and the platinum ions—water material is used in a second set of electrochemical steps to form bi-metallic Au—Pt nanocrystals in suspension.
- Specifically, in a preferred embodiment, a second set of steps of the electrochemical process involves the nucleation and growth of bi-metallic nanocrystals, such growth including: (1) mixtures of two metals, (2) alloys of two metals and/or (3) heteroaggregates (e.g., composites) of two metals. For example, the platinum species and water output from the first steps of the preferred embodiment (note that electrochemical processing enhancer used during the first electrochemical processing is also present) act as raw material input into the second electrochemical processing steps of a preferred embodiment. Depending on the particular concentrations and type of formed platinum species, processing enhancer(s) components, raw material and run conditions of the electrochemical processes (including devices used), one or more of the aforementioned bi-metallic nanocrystalline components can be produced as stable nanocrystals in the aqueous suspension during the second set of electrochemical processing steps.
- Because the grown bi-metallic nanocrystals have “bare” or “clean” surfaces of gold and/or platinum metal (e.g., in the zero oxidation state) bi-metallic nanocrystal surfaces are highly catalytic or are highly biocatalytic (as well as highly bioavailable). The bi-metallic nanocrystals are essentially surrounded by a water-based jacket comprising, for example, water species which are made available due to, for example, lysing of the water which occurs in one or more steps of a preferred embodiment. The lysed species may include hydrated electrons, OH−, H*, H3O, H2O2, etc. However, without wishing to be bound by any particular theory or explanation, OH− groups (e.g., from either lysed water or processing enhancer) may locate themselves around the formed bi-metallic crystals and create an electrostatic interaction therewith. These clean surface features provide novel and enhanced performance in a variety of industrial and medical applications and/or can result in decreased general undesirable toxicity in medical applications because no undesirable toxins or poisons are present on the surfaces due to the manufacturing process.
- In a preferred embodiment, the nanocrystals are not dried before use but instead are directly used in the liquid they were formed in (i.e., forming a suspension). Alternatively, the formed suspensions can be formed into a concentrate or a reconstituted concentrate thereof. It appears that completely removing these crystals from their suspension liquid (e.g., completely drying) may, in certain cases, adversely affect the surface properties of the crystals, (e.g., partial oxidation may occur, the stabilizing groups may be irreparably damaged, etc.) and/or may adversely affect the ability to rehydrate the crystals. For example, if the initially formed water jacket includes OH− which assist in electrostatic interactions, then changing the OH− coordination may upset the stability of the suspension.
- However, it has been discovered that a certain concentration process utilizing a dialysis procedure can be used. The dialysis procedure involves placement of the formed bi-metallic nanocrystal suspension inside of a dialysis bag. A polyethylene solution is located on the outside of the dialysis bag (e.g., the dialysis bag can be placed with a suitable container housing polyethylene glycol (PEG)) permits water to be removed from the formed bi-metallic nanocrystal suspension by osmotic pressure without comprising the stability of the nanocrystals in suspension. Further, if certain ionic constituents remain in the liquid which suspends the nanocrystals, some or all of such ionic constituents can be removed from such liquid, if desired, so long as such removal does not adversely affect the stability and/or performance of the bi-metallic nanocrystals or nanocrystal suspension.
- Further, for some medical-based products, it may be optimal to use sterile pharmaceutical grade water (e.g., USP) or the like in addition to the aforementioned process enhancers used in the manufacturing processes. In some cases, the water could be even more pure than USP by using reverse osmosis and/or ionic filtration means.
- Alternatively, in another embodiment, the bi-metallic nanocrystals may be dried in situ into/onto, for example, an electrode or substrate which takes part in another reaction such as another electrochemical, chemical or catalytic process. For example, the bi-metallic nanocrystals made according to this invention can also be used for industrial applications where metal reactivity is important (e.g., catalytic and/or electrochemical processes) but where pharmaceutical grade products/ingredients are not required. When prepared for non-pharmaceutical uses, the bi-metallic nanocrystals can be made in a wider variety of solvents and with a wider variety of process enhancers, as discussed herein, depending on the specifc application. However, the clean aspects of the bi-metallic nanocrystal surfaces should be preserved to achieve superior performance.
- In another preferred embodiment of the invention, the electrochemical process steps of the invention can be controlled so as to result in more than one type of bi-metallic nanocrystal being present in the resultant suspension. For example, mixtures of platinum and gold nanocrystals may exist in suspension, alloys of platinum and gold nanocrystals may exist in suspension and/or nanocrystal heteroaggregates of platinum and gold may also exist in suspension.
- According to the processes herein, the bi-metallic nanocrystals can be grown in a manner that provides unique and identifiable surface characteristics such as spatially extended low index, crystal planes {111}, {110} and/or {100} and groups of such planes (and their equivalents). Such crystal planes can show different and desirable catalytic performances. A variety of crystalline shapes can be found in bi-metallic nanoparticle suspensions made according to embodiments disclosed herein. Further, the surfaces of bi-metallic nanocrystals grown should be highly active due to their crystalline condition (e.g., surface defects) as well as being clean.
- Any desired average size of bi-metallic nanocrystals below 100 nm can be achieved. The most desirable nanocrystalline size ranges include those having an average crystal size (as measured and determined by specific techniques disclosed in detail herein) that is predominantly less than 100 nm, and more typically less than 50 nm, even more typically less than 30 nm, and in many of the preferred embodiments disclosed herein, the mode for the nanocrystal size distribution is less than 20 nm and within an even more preferable range of 8-18 nm. However, for some applications, the techniques of the invention can be used to manufacture much larger particles.
- Resulting bi-metallic nanocrystalline suspensions or colloids can be provided that have or are adjusted to have target pH ranges. When prepared with, for example, a sodium bicarbonate or other “basic” (e.g., one where the OH− concentration is caused to be relatively high) process enhancer, in the amounts disclosed in detail herein, the pH range is typically 8-11, which can be adjusted as desired. Still further, the use of certain processing enhancers can result in even higher pH ranges, such as a pH of about 9-12 or even 10.3-12.0.
- The nature and/or amount of the surface charge (i.e., positive or negative) on formed bi-metallic nanocrystals can have a large influence on the behavior and/or effects of the nanocrystal/suspension or colloid (or the concentrated nanocrystals). For example, for biomedical applications, protein coronas such as albumin coronas and/or transferrin coronas formed in vivo can be influenced by surface charge or surface characteristics (e.g., including impurities or residual components present from processing techniques) of a nanoparticle. Such coronas dictate the biological identity of the nanoparticle and thus direct biologic availability.
- Such surface charges are commonly referred to as “zeta potential”. It is known that the larger the zeta potential (either positive or negative), the greater the stability of the nanoparticles in the solution (i.e., the suspension is more stable). By controlling the nature and/or amount of the surface charges of formed nanoparticles or nanocrystals, the performance of such nanoparticle suspensions can be controlled in biological and non-biological applications.
- Zeta potential is known as a measure of the electo-kinetic potential in colloidal systems and is also referred to as surface charge on particles. Zeta potential is the potential difference that exists between the stationary layer of fluid and the fluid within which the particle is dispersed. A zeta potential is often measured in millivolts (i.e., mV). The zeta potential value of approximately 20-25 mV is an arbitrary value that has been chosen to determine whether or not a dispersed particle is stable in a dispersion medium. Thus, when reference is made herein to “zeta potential”, it should be understood that the zeta potential referred to is a description or quantification of the magnitude of the electrical charge present at the double layer.
- The zeta potential is calculated from the electrophoretic mobility by the Henry equation:
-
- where z is the zeta potential, UE is the electrophoretic mobility, ε is a dielectric constant, η is a viscosity, f(ka) is Henry's function. For Smoluchowski approximation f(ka)=1.5.
- Zeta potentials (“ZP”) for the bi-metallic nanocrystals prepared according the methods herein typically have a ZP of at least −20 mV, more typically at least about −30 mV, even more typically, at least about −40 mV and even more typically at least about −50 mV.
- Further, another important aspect of the preferred embodiments is that the raw material metal ions are produced by the donor electrode metals of Pt and Au (e.g., sacrificial or donor electrodes) due to the processing conditions of the preferred embodiments. This “top-down” first set of electrochemical steps means that materials typically used to make metal-based nanoparticles in other techniques, such as metal salts (e.g., Pt salts, Au salts, etc.) are not required to be used in the embodiments disclosed herein. Thus, other constituents (which can be undesirable) of the metal salts, such as Cl− or various chlorine-based materials, do not occur, or are not a required part of a product made according to the preferred embodiments herein. In other words, for example, the other constituents that comprise various metal-based raw material salts do not need to be present in the bi-metallic nanocrystal suspensions discussed herein (e.g., bi-metallic suspensions can be chlorine or chloride-free). Of course, it should be noted that the presence of chlorine-based materials dissolved in the suspension, and were not required or essential to the nanoparticle production process, are contemplated as being within the metes and bounds of this disclosure.
- A set of novel process steps is provided to produce these unique bi-metallic nanocrystals. The process steps involve the creation of the bi-metallic nanocrystals in water. In a preferred embodiment, the water contains an added “process enhancer” which does not significantly bind to the formed nanocrystals, but rather facilitates nucleation/crystal growth during the electrochemical-stimulated growth process. The process enhancer serves important roles in the process including providing charged ions in the electrochemical solution to permit the crystals to be grown. These novel electrochemical processes can occur in either a batch, semi-continuous or continuous process. These processes result in controlled bi-metallic nanocrystalline concentrations of gold and platinum, controlled bi-metallic nanocrystal sizes and controlled bi-metallic nanocrystal size ranges. Novel manufacturing assemblies are provided to produce these bi-metallic nanocrystals. In another embodiment, metallic-based constituents, such as desirable metallic ions, can be included separately or combined with bi-metallic nanocrystal suspensions.
- In one preferred embodiment, the bi-metallic nanocrystal suspensions or colloids are made or grown by electrochemical techniques in either a batch, semi-continuous or continuous process, wherein the amount, average particle size, crystal plane(s) and/or particle shape(s) and/or particle shape distributions are controlled and/or optimized to achieve high biological activity and low cellular/biologic toxicity (e.g., a high therapeutic index). Desirable average crystal sizes include a variety of different ranges, but the most desirable ranges include average crystal sizes that are predominantly less than 100 nm and more typically, for many uses, less than 50 nm and even more typically for a variety of, for example, oral uses, less than 30 nm, and in many of the preferred embodiments disclosed herein, the mode for the nanocrystal size distribution is less than 20 nm and within an even more preferable range of 2-18 nm, as measured by a zetasizer (as described in more detail herein). Further, the particles desirably contain crystal planes, such desirable (and often highly reactive) crystal planes, include crystals having {111}, {110} and/or {100} facets, as well as defects, which can result in superior interactions such as catalytic.
- Further, by following the inventive electrochemical manufacturing processes of the invention, these bi-metallic nanocrystals can be alloys, or can be combined with other metals in liquids such that metal “coatings” may occur on other metals to form composites or heteroaggregates or alternatively, mixtures of metal-based nanocrystals can be made.
- Still further, bi-metallic nanocrystal suspensions or colloids of the present invention can be mixed or combined with other metallic-based solutions or colloids to form novel solutions or colloid mixtures (e.g., in this instance, distinct metal species can still be discerned).
- Methods for making novel metallic-based nanocrystal suspensions or colloids according to the invention relate generally to novel methods and novel devices for the continuous, semi-continuous and batch manufacture of a variety of constituents in a liquid including micron-sized particles, nanocrystals, ionic species and aqueous-based compositions of the same, including, nanocrystal/liquid(s), solution(s), colloid(s) or suspension(s). The constituents and bi-metallic nanocrystals produced can comprise a variety of possible compositions, concentrations, sizes, crystal planes (e.g., spatially extended low index crystal planes) and/or shapes, which together can cause the inventive compositions to exhibit a variety of novel and interesting physical, catalytic, biocatalytic and/or biophysical properties. The liquid(s) used and created/modified during the process can play an important role in the manufacturing of, and/or the functioning of the constituents (e.g., nanocrystals) independently or synergistically with the liquids which contain them. The particles (e.g., nanocrystals) are caused to be present (e.g., created and/or the liquid is predisposed to their presence (e.g., conditioned)) in at least one liquid (e.g., water) by, for example, typically utilizing at least one adjustable plasma (e.g., created by at least one AC and/or DC power source), which adjustable plasma communicates with at least a portion of a surface of the liquid. However, effective constituent (e.g., nanocrystals) suspensions or colloids can be achieved without the use of such plasmas as well.
- Gold and platinum-based electrodes of various composition(s) and/or unique configurations or arrangements are preferred for use in the formation of the adjustable plasma(s). Utilization of at least one subsequent and/or substantially simultaneous adjustable electrochemical processing technique is also preferred. Gold and platinum-based electrodes are preferred for use in the electrochemical processing technique(s). Electric fields, magnetic fields, electromagnetic fields, electrochemistry, pH, zeta potential, chemical/crystal constituents present, etc., are just some of the variables that can be positively affected by the adjustable plasma(s) and/or adjustable electrochemical processing technique(s) of the invention. Multiple adjustable plasmas and/or adjustable electrochemical techniques are preferred in many embodiments of the invention to achieve many of the processing advantages of the present invention, as well as many of the novel bi-metallic nanocrystals and bi-metallic nanocrystal compositions which result from practicing the teachings of the preferred embodiments to make an almost limitless set of inventive aqueous solutions, suspensions and/or colloids.
- In the continuous process preferred embodiments of the invention, at least one liquid, for example water, flows into, through and out of at least one first trough member and such liquid is processed, conditioned, modified and/or effected by said at least one adjustable plasma and/or said at least one adjustable electrochemical technique. The results of the continuous processing in the first trough member include new constituents in the liquid, such as ionic constituents, nanocrystals (e.g., platinum-based nanocrystals) of novel and/or controllable size, hydrodynamic radius, concentration, crystal sizes and crystal size ranges, zeta potential, pH and/or properties, such platinum nanocrystal/ion/liquid mixture being produced in an efficient and economical manner.
- Further, in a preferred embodiment, a first set of steps of the process involves the in situ creation of platinum species (e.g., raw materials) from a platinum metal source. The platinum species is created in water which contains a “process enhancer” or “processing enhancer” (typically an inorganic material or carbonate or such) which does not significantly bind to the formed nanocrystals in suspension, but rather facilitates removal of metal ions from a donor metal source, and/or assists in nucleation/growth during electrochemical-stimulated nanocrystal growth processes. More specifically, the process enhancer serves important roles in the process including providing charged ions in the electrochemical solution to permit the nanocrystals to be grown. The process enhancer is critically a compound(s) which remains in solution, and/or does not form a coating (e.g., an organic coating), and/or does not adversely affect the performance of the formed nanocrystals or the formed suspension(s) (e.g., is inert), and/or can be destroyed, evaporated, removed or otherwise lost during one or more steps of the electrochemical process. A preferred process enhancer is sodium bicarbonate. Examples of other process enhancers are sodium carbonate, potassium bicarbonate, potassium carbonate, trisodium phosphate, disodium phosphate, monosodium phosphate, potassium phosphates or the like and combinations thereof. Another particularly preferred processing enhancer is a mixture of sodium bicarbonate and potassium hydroxide.
- Desirable concentration ranges for the processing enhancer include typically 0.01-20 grams/gallon (0.0026-2.1730 mg/ml), more typically, 0.1-7.5 grams/gallon (0.0264-1.9813 mg/ml) and most typically, 0.5-2.0 grams/gallon (0.13210-0.5283 mg/ml).
- In a preferred embodiment, a second set of steps of the process involves the nucleation and growth of bi-metallic-based nanocrystals, such growth being: (1) mixtures of two metals, (2) alloys of two metals and/or (3) heteroaggregates of two metals. For example, the aqueous output from the first steps of the preferred embodiment containing water, platinum species resulting from the first steps of the process, and processing enhancer used during the first set of steps, acts as raw material input into the second electrochemical steps of a preferred embodiment. Depending on the particular concentrations of platinum species, processing enhancer(s) constituent(s) and run conditions of the electrochemical processes (including devices used), one or more of the aforementioned bi-metallic nanocrystalline components can be produced as stable bi-metallic nanocrystals in the aqueous suspension during the second set of steps.
- Certain processing enhancers may dissociate into positive ions (cations) and negative ions (anions). The anions and/or cations, depending on a variety of factors including liquid composition, concentration of ions, change state of ions, applied fields, frequency of applied fields, waveform of the applied filed, temperature, pH, zeta potential, etc., will navigate or move toward oppositely charged electrodes. When said ions are located at or near such electrodes, the ions may take part in one or more reactions with the electrode(s) and/or other constituent(s) located or created at or near such electrode(s). Sometimes ions may react with one or more materials in the electrode. Such reactions may be desirable in some cases or undesirable in others. Further, sometimes ions present in a solution between electrodes may not react to form a product, but rather may influence material in the electrode (or near the electrode) to form metallic nano-crystals that are “grown” from material provided by the donor electrode. For example, certain metal ions may enter the liquid 3 from the
electrode 5 and be caused to come together (e.g., nucleate) to form constituents (e.g., ions, nanocrystals, etc.) within theliquid 3. - Further, it is important to select a process enhancer that will not negatively impact performance such as, for example, impart negative performance or, for example, toxicity to the bi-metallic nanocrystal, or to the liquid that the crystal is suspended in, to maximize acceptability for various commercial uses (e.g., pharmaceutical, catalytic, medical diagnostic, etc). For example, for certain applications, chlorine ions or chlorides or chlorine-based materials may be undesired if such species create, for example, gold chloride salts, which may be undesirable for several reasons (e.g., may affect toxicity, stability, etc.).
- Additionally, certain processing enhancers that involve hydroxyl groups OH− (e.g., which are part of the processing enhancer or result from addition of processing enhancers to the liquid 3) can also be desirable. In this regard, desirable processing enhancers of NaOH, KOH and NaHCO3 (and mixtures of the same) are specifically disclosed as being desirable in some preferred embodiments herein.
- Further, depending upon the specific formed products, drying, concentrating and/or freeze drying can also be utilized to remove at least a portion of, or substantially all of, the suspending liquid, resulting in, for example, partially or substantially completely dehydrated bi-metallic nanocrystals. If such nanocrystals are ultimately located on a substrate (e.g., a catalysis substrate or an electrode) complete drying may be required. If solutions, suspensions or colloids are completely dehydrated, the metal-based species, in some cases, should be capable of being rehydrated by the addition of liquid (e.g., of similar or different composition than that which was removed). However, not all compositions/colloids of the present invention can be completely dehydrated without adversely affecting performance of the composition/colloid. For example, many nanocrystals formed in a liquid tend to clump or stick together (or adhere to surfaces) when dried. If such clumping is not reversible during a subsequent rehydration step, dehydration should be avoided. However, for a variety of applications such clumping may be acceptable. Further, when drying on a substrate, such clumping may be avoided.
- In general, it is possible to concentrate, several fold, certain solutions, suspensions or colloids of bi-metallic nanocrystals made according to the invention, without destabilizing the composition. For example, without wishing to be bound, if the initially formed water jacket includes OH− which assist in electrostatic interactions, then changing the OH− coordination in any way may upset the stability of the suspension.
- However, it has been discovered that a certain concentration process utilizing a dialysis procedure can be used. The dialysis procedure involves placement of the formed bi-metallic nanocrystal suspension inside of a dialysis bag. A polyethylene solution is located on the outside of the dialysis bag (e.g., the dialysis bag can be placed with a suitable container holding polyethylene glycol (PEG)) and water can be removed from the formed bi-metallic nanocrystal suspension by osmotic pressure without comprising the stability of the nanocrystals in suspension. Further, if certain ionic constituents remain in the liquid which suspends the nanocrystals, some or all of such ionic constituents can be removed from such liquid, so long as such removal does not adversely affect the stability and/or performance of the bi-metallic nanocrystals or nanocrystal suspension.
- While the following discussion is believed to be complete, the reader is also directed to a related application, International Publication No. WO/2011/006007 published on 13 Jan. 2011, the subject matter of which is expressly incorporated herein by reference.
- One important aspect of the invention involves the creation of at least one adjustable plasma, which adjustable plasma is located between at least one electrode positioned adjacent to (e.g., above) at least a portion of the surface of a liquid (e.g., water) and at least a portion of the surface of the liquid itself. The liquid is placed into electrical communication with at least one second electrode (or a plurality of second electrodes) causing the surface of the liquid to function as an electrode, thus taking part in the formation of the adjustable plasma. This configuration has certain characteristics similar to a dielectric barrier discharge configuration, except that the surface of the liquid is an active electrode participant in this configuration.
- Each adjustable plasma utilized can be located between the at least one electrode located above a surface of the liquid and a surface of the liquid due to at least one electrically conductive electrode being located somewhere within (e.g., at least partially within) the liquid. At least one power source (in a preferred embodiment, at least one source of volts and amps such as a transformer or power source) is connected electrically between the at least one electrode located above the surface of the liquid and the at least one electrode contacting the surface of the liquid (e.g., located at least partially, or substantially completely, within the liquid). The electrode(s) may be of any suitable composition (however, platinum and gold are preferred) and suitable physical configuration (e.g., size and shape) which results in the creation of a desirable plasma between the electrode(s) located above the surface of the liquid and at least a portion of the surface of the liquid itself.
- The applied power (e.g., voltage and amperage) between the electrode(s) (e.g., including the surface of the liquid functioning as at least one electrode for forming the plasma) can be generated by any suitable source (e.g., voltage from a transformer) including both AC and DC sources and variants and combinations thereof. Generally, the electrode or electrode combination located within (e.g., at least partially below the surface of the liquid) takes part in the creation of a plasma by providing voltage and current to the liquid or solution. However, the adjustable plasma is actually located between at least a portion of the electrode(s) located above the surface of the liquid (e.g., at a tip or point thereof) and one or more portions or areas of the liquid surface itself. In this regard, the adjustable plasma can be created between the aforementioned electrodes (i.e., those located above at least a portion of the surface of the liquid and a portion of the liquid surface itself) when a breakdown voltage of the gas or vapor around and/or between the electrode(s) and the surface of the liquid is achieved or maintained.
- In one embodiment of the invention, the liquid comprises water (or water containing certain processing enhancer(s)), and the gas between the surface of the water and the electrode(s) above the surface of the water (i.e., that gas or atmosphere that takes part in the formation of the adjustable plasma) comprises air. The air can be controlled to contain various different water content(s) or a desired humidity which can result in different compositions, concentrations, crystal size distributions and/or crystal shape distributions of constituents (e.g., nanocrystals) being produced according to the present invention (e.g., different amounts of certain constituents in the adjustable plasma and/or in the solution or suspension can be a function of the water content in the air located above the surface of the liquid) as well as different processing times required to obtain certain concentrations of various constituents in the liquid, etc.
- The breakdown electric field at standard pressures and temperatures for dry air is about 3 MV/m or about 30 kV/cm. Thus, when the local electric field around, for example, a metallic point exceeds about 30 kV/cm, a plasma can be generated in dry air. Equation (1) gives the empirical relationship between the breakdown electric field “Ec” and the distance “d” (in meters) between two electrodes:
-
- Of course, the breakdown electric field “Ec” will vary as a function of the properties and composition of the gas or vapor located between electrodes. In this regard, in one preferred embodiment where water (or water containing a processing enhancer) is the liquid, significant amounts of water vapor can be inherently present in the air between the “electrodes” (i.e., between the at least one electrode located above the surface of the water and the water surface itself which is functioning as one electrode for plasma formation) and such water vapor should have an effect on at least the breakdown electric field required to create a plasma therebetween. Further, a higher concentration of water vapor can be caused to be present locally in and around the created plasma due to the interaction of the adjustable plasma with the surface of the water. The amount of “humidity” present in and around the created plasma can be controlled or adjusted by a variety of techniques discussed in greater detail later herein. Likewise, certain components present in any liquid can form at least a portion of the constituents forming the adjustable plasma located between the surface of the liquid and the electrode(s) located adjacent (e.g., along) the surface of the liquid. The constituents in the adjustable plasma, as well as the physical properties of the plasma per se, can have a dramatic influence on the liquid, as well as on certain of the processing techniques (discussed in greater detail later herein).
- The electric field strengths created at and near the electrodes are typically at a maximum at a surface of an electrode and typically decrease with increasing distance therefrom. In cases involving the creation of an adjustable plasma between a surface of the liquid and the at least one electrode(s) located adjacent to (e.g., above) the liquid, a portion of the volume of gas between the electrode(s) located above a surface of a liquid and at least a portion of the liquid surface itself can contain a sufficient breakdown electric field to create the adjustable plasma. These created electric fields can influence, for example, behavior of the adjustable plasma, behavior of the liquid (e.g., influence the crystal state of the liquid) behavior of constituents in the liquid, etc.
- In this regard,
FIG. 1 shows one embodiment of apoint source electrode 1 having a triangular cross-sectional shape located a distance “x” above thesurface 2 of a liquid 3 flowing, for example, in the direction “F”. Anadjustable plasma 4 can be generated between the tip orpoint 9 of theelectrode 1 and thesurface 2 of theliquid 3 when anappropriate power source 10 is connected between thepoint source electrode 1 and theelectrode 5, which electrode 5 communicates with the liquid 3 (e.g., is at least partially below thesurface 2 of the liquid 3). - The
adjustable plasma region 4, created in the embodiment shown inFIG. 1 can typically have a shape corresponding to a cone-like structure or an ellipsoid-like structure, for at least a portion of the process, and in some embodiments of the invention, can maintain such shape (e.g., cone-like shape) for substantially all of the process. The volume, intensity, constituents (e.g., composition), activity, precise locations, etc., of the adjustable plasma(s) 4 will vary depending on a number of factors including, but not limited to, the distance “x”, the physical and/or chemical composition of the electrode 1, the shape of the electrode 1, the power source 10 (e.g., DC, AC, rectified AC, the applied polarity of DC and/or rectified AC, AC or DC waveform, RF, etc.), the power applied by the power source (e.g., the volts applied, which is typically 1000-5000 Volts, and more typically 1000-1500 Volts, the amps applied, electron velocity, etc.) the frequency and/or magnitude of the electric and/or magnetic fields created by the power source applied or ambient, electric, magnetic or electromagnetic fields, acoustic fields, the composition of the naturally occurring or supplied gas or atmosphere (e.g., air, nitrogen, helium, oxygen, ozone, reducing atmospheres, etc.) between and/or around the electrode 1 and the surface 2 of the liquid 3, temperature, pressure, volume, flow rate of the liquid 3 in the direction “F”, spectral characteristics, composition of the liquid 3, conductivity of the liquid 3, cross-sectional area (e.g., volume) of the liquid near and around the electrodes 1 and 5, (e.g., the amount of time (i.e., dwell time) the liquid 3 is permitted to interact with the adjustable plasma 4 and the intensity of such interactions), the presence of atmosphere flow (e.g., air flow) at or near the surface 2 of the liquid 3 (e.g., fan(s) or atmospheric movement means provided) etc., (discussed in more detail later herein). - The composition of the electrode(s) 1 involved in the creation of the adjustable plasma(s) 4 of
FIG. 1 , in one preferred embodiment of the invention, are metal-based compositions (e.g., metals such as gold, platinum and/or alloys or mixtures thereof, etc.), but theelectrodes plasma 4 in, for example, air above thesurface 2 of a liquid 3 (e.g., water) will, typically, produce at least some ozone, as well as amounts of nitrogen oxide and other components. These produced components can be controlled and may be helpful or harmful to the formation and/or performance of the resultant constituents in the liquid (e.g., nanocrystals) and/or, nanocrystal suspensions or colloids produced and may need to be controlled by a variety of different techniques. As shown inFIG. 1 , theadjustable plasma 4 actually contacts thesurface 2 of theliquid 3. In this embodiment of the invention, material (e.g., metal) from theelectrode 1 may comprise a portion of the adjustable plasma 4 (e.g., and thus be part of the emission spectrum of the plasma) and may be caused, for example, to be “sputtered” onto and/or into the liquid 3 (e.g., water). Accordingly, when metal(s) are used as the electrode(s) 1, a variety of constituents can be formed in the electrical plasma, resulting in certain constituents becoming part of the processing liquid 3 (e.g., water), including, but not limited to, elementary metal(s), metal ions, Lewis acids, Bronsted-Lowry acids, metal oxides, metal nitrides, metal hydrides, metal hydrates and/or metal carbides, etc., can be found in the liquid 3 (e.g., for at least a portion of the process and may be capable of being involved in simultaneous/subsequent reactions), depending upon the particular set of operating conditions associated with theadjustable plasma 4 and/or subsequent electrochemical processing operations. Such constituents may be transiently present in theprocessing liquid 3 or may be semi-permanent or permanent. If such constituents are transient or semi-permanent, then the timing of subsequent reactions (e.g., electrochemical reactions) with such formed constituents can influence final products produced. If such constituents are permanent, they should not adversely affect the desired performance of the active ingredient nanocrystals. - Further, depending on, for example, electric, magnetic and/or electromagnetic field strength in and around the
liquid 3 and the volume ofliquid 3 exposed to such fields, the physical and chemical construction of the electrode(s) 1 and 5, atmosphere (naturally occurring or supplied), liquid composition, greater or lesser amounts of electrode(s) materials(s) (e.g., metal(s) or derivatives of metals) may be found in theliquid 3. In certain situations, the material(s) (e.g., metal(s) or metal(s) composite(s)) or constituents (e.g., Lewis acids, Bronsted-Lowry acids, etc.) found in the liquid 3 (permanently or transiently), or in theplasma 4, may have very desirable effects, in which case relatively large amounts of such materials will be desirable; whereas in other cases, certain materials found in the liquid 3 (e.g., by-products) may have undesirable effects, and thus minimal amounts of such materials may be desired in the liquid-based final product. Accordingly, electrode composition can play an important role in the materials that are formed according to the embodiments disclosed herein. The interplay between these components of the invention are discussed in greater detail later herein. - Still further, the electrode(s) 1 and 5 may be of similar chemical composition (e.g., have the same chemical element as their primary constituent) and/or mechanical configuration or completely different compositions (e.g., have different chemical elements as their primary constituent) in order to achieve various compositions and/or structures of liquids and/or specific effects discussed later herein.
- The distance “y” between the electrode(s) 1 and 5; or 1 and 1 (shown later herein) or 5 and 5 (shown later herein) is one important aspect of the invention. In general, when working with power sources capable of generating a plasma under the operating condition, the location of the smallest distance “y” between the closest portions of the electrode(s) used in the present invention should be greater than the distance “x” in order to prevent an undesirable arc or formation of an unwanted corona or plasma occurring between the electrode (e.g., the electrode(s) 1 and the electrode(s) 5) (unless some type of electrical insulation is provided therebetween). Features of the invention relating to electrode design, electrode location and electrode interactions between a variety of electrodes are discussed in greater detail later herein.
- The power applied through the
power source 10 may be any suitable power which creates a desirableadjustable plasma 4 under all of the process conditions of the present invention. In one preferred mode of the invention, an alternating current from a step-up transformer is utilized. Preferred transformer(s) 60 (see e.g.,FIGS. 7 a-7 b ) for use in various embodiments disclosed herein, have deliberately poor output voltage regulation made possible by the use of magnetic shunts in thetransformer 60. Thesetransformers 60 are known as neon sign transformers. This configuration limits current flow into the electrode(s) 1/5. With a large change in output load voltage, thetransformer 60 maintains output load current within a relatively narrow range. - The
transformer 60 is rated for its secondary open circuit voltage and secondary short circuit current. Open circuit voltage (OCV) appears at the output terminals of thetransformer 60 only when no electrical connection is present. Likewise, short circuit current is only drawn from the output terminals if a short is placed across those terminals (in which case the output voltage equals zero). However, when a load is connected across these same terminals, the output voltage of thetransformer 60 should fall somewhere between zero and the rated OCV. In fact, if thetransformer 60 is loaded properly, that voltage will be about half the rated OCV. - The
transformer 60 is known as a Balanced Mid-Point Referenced Design (e.g., also formerly known as balanced midpoint grounded). This is most commonly found in mid to higher voltage rated transformers and most 60 mA transformers. This is the only type transformer acceptable in a “mid-point return wired” system. The “balanced”transformer 60 has oneprimary coil 601 with twosecondary coils 603, one on each side of the primary coil 601 (as shown generally in the schematic view inFIG. 7 b ). Thistransformer 60 can in many ways perform like two transformers. Just as the unbalanced midpoint referenced core and coil, one end of eachsecondary coil 603 is attached to thecore 602 and subsequently to the transformer enclosure and the other end of the eachsecondary coil 603 is attached to an output lead or terminal. Thus, with no connector present, an unloaded 15,000 volt transformer of this type, will measure about 7,500 volts from each secondary terminal to the transformer enclosure but will measure about 15,000 volts between the two output terminals. Theseexemplary transformers 60 were utilized to form theplasmas 4 disclosed in the Examples herein. However, other suitable transformers (or power sources) should also be understood as falling within the metes and bounds of the invention. However, a different power supply 501AC (discussed elsewhere herein) is utilized for theelectrodes 5/5′ in most of the other examples disclosed herein. - In further reference to the configurations shown in
FIG. 1 ,electrode holders electrode holders portions portions portions electrode holders member 8 can be made of any suitable material which prevents undesirable electrical events (e.g., arcing, melting, etc.) from occurring, as well as any material which is structurally and environmentally suitable for practicing the present invention. Typical materials include structural plastics such as polycarbonates, plexiglass (poly (methyl methacrylate), polystyrene, acrylics, and the like. Additional suitable materials for use with the present invention are discussed in greater detail elsewhere herein. - Preferred techniques for automatically raising and/or lowering the
electrodes power source 10 can be connected in any convenient electrical manner to theelectrodes wires electrode holders portions portions electrodes -
FIG. 2 shows another schematic of a preferred embodiment of the invention, wherein acontrol device 20 is connected to theelectrodes control device 20 remotely (e.g., upon command from another device or component) raises and/or lowers theelectrodes surface 2 of theliquid 3. Thecontrol device 20 is discussed in more detail later herein. In this one preferred aspect of the invention, theelectrodes FIG. 2 ) containing an appropriate software control program. Accordingly, the embodiment shown inFIG. 1 should be considered to be a manually controlled apparatus for use with the techniques of the present invention, whereas the embodiment shown inFIG. 2 should be considered to include an automatic apparatus orassembly 20 which can remotely raise and lower theelectrodes FIG. 2 preferred embodiments of the invention can also employ computer monitoring and computer control of the distance “x” of thetips 9 of the electrodes 1 (andtips 9′ of the electrodes 5) away from thesurface 2; or computer monitoring and/or controlling the rate(s) which theelectrode 5 is advanced into/through theliquid 3. Thus, the appropriate commands for raising and/or lowering theelectrodes FIG. 2 ). -
FIGS. 3 a-3 e show perspective views of various desirable electrode configurations for theelectrode 1 shown inFIGS. 1-2 (as well as in other Figures and embodiments discussed later herein). The electrode configurations shown inFIGS. 3 a-3 e are representative of a number of different configurations that are useful in various embodiments of the present invention. Criteria for appropriate electrode selection for theelectrode 1 include, but are not limited to the following conditions: the need for a very well defined tip orpoint 9, composition, mechanical limitations, the ability to make shapes from the material comprising theelectrode 1, conditioning (e.g., heat treating or annealing) of the material comprising theelectrode 1, convenience, the constituents introduced into theplasma 4, the influence upon theliquid 3, etc. In this regard, a small mass of material comprising theelectrodes 1 shown in, for example,FIGS. 1-2 may, upon creation of theadjustable plasmas 4 according to the present invention (discussed in greater detail later herein), rise to operating temperatures where the size and or shape of the electrode(s) 1 can be adversely affected. In this regard, for example, if theelectrode 1 was of relatively small mass (e.g., if the electrode(s) 1 was made of gold and weighed about 0.5 gram or less) and included a very fine point as thetip 9, then it is possible that under certain sets of conditions used in various embodiments herein that a fine point (e.g., a thin wire having a diameter of only a few millimeters and exposed to a few hundred to a few thousand volts; or a triangular-shaped piece of metal) would be incapable of functioning as the electrode 1 (e.g., theelectrode 1 could deform undesirably or melt), absent some type of additional interactions (e.g., internal cooling means or external cooling means such as a fan, etc.). Accordingly, the composition of (e.g., the material comprising) the electrode(s) 1 may affect possible suitable electrode physical shape due to, for example, melting points, pressure sensitivities, environmental reactions (e.g., the local environment of theadjustable plasma 4 could cause undesirable chemical, mechanical and/or electrochemical erosion of the electrode(s)), etc. - Moreover, it should be understood that in alternative preferred embodiments of the invention, well defined sharp points are not always required for the
tip 9. In this regard, theelectrode 1 shown inFIG. 3 e comprises arounded tip 9. It should be noted that partially rounded or arc-shaped electrodes can also function as theelectrode 1 because theadjustable plasma 4, which is created in the inventive embodiments shown herein (see, for example,FIGS. 1-2 ), can be created from rounded electrodes or electrodes with sharper or more pointed features. During the practice of the inventive techniques of the present invention, such adjustable plasmas can be positioned or can be located along various points of theelectrode 1 shown inFIG. 3 e . In this regard,FIG. 4 shows a variety of points “a-g” which correspond to initiatingpoints 9 for theplasmas 4 a-4 g which occur between theelectrode 1 and thesurface 2 of theliquid 3. Accordingly, it should be understood that a variety of sizes and shapes corresponding toelectrode 1 can be utilized in accordance with the teachings of the present invention. Still further, it should be noted that thetips electrodes electrode tips - The electrode configurations shown generally in
FIGS. 1 and 2 can create different results (e.g., different conditioning effects for the fluid 3, different pH's in the fluid 3, different nanocrystals sizes and size distribution, different nanocrystal shapes and nanocrystal shape distributions, and/or amounts of constituents (e.g., nanocrystal matter and/or metal ions from the donor electrode(s)) found in the fluid 3, different functioning of the fluid/nanocrystal combinations (e.g., different biologic/biocatalytic effects), different zeta potentials, etc.) as a function of a variety of features including the electrode orientation and position relative to the fluid flow direction “F”, cross-sectional shape and size of the trough member 30 (or 30 a′ and/or 30 b′), and/or amount of the liquid 3 within the trough member 30 and/or rate of flow of the liquid 3 within the trough member 30 and in/around the electrodes 5 a/5 b, the thickness of the electrodes, the number of electrode pairs provided and their positioning in the trough member 30 relative to each other as well as their depth into the liquid 3 (i.e., amount of contact with the liquid 3), the rate of movement of the electrodes into/through the liquid 3 (which maintains or adjusts the surface profile or shape if the electrodes), the power applied to the electrode pairs, etc. Further, the electrode compositions, size, specific shape(s), number of different types of electrodes provided, voltage applied, amperage applied and/or achieved within theliquid 3, AC source (and AC source frequency and AC waveform shape, duty cycle, etc.), DC source, RF source (and RF source frequency, duty cycle, etc.), electrode polarity, etc., can all influence the properties of the liquid 3 (and/or the nanocrystals formed or contained in the liquid 3) as the liquid 3 contacts, interacts with and/or flows past theseelectrodes -
FIGS. 5 a-5 e show cross-sectional views of the liquid containingtrough member 30 used in preferred embodiments herein. The distance “S” and “S′” for the preferred embodiment shown in each ofFIGS. 5 a-5 e measures, for example, between about 0.25″ and about 6″ (about 0.6 cm-15 cm). The distance “M” ranges from about 0.25″ to about 6″ (about 0.6 cm-15 cm). The distance “R” ranges from about ½″ to about 7″ (about 1.2 cm to about 17.8 cm). All of these embodiments (as well as additional configurations that represent alternative embodiments are within the metes and bounds of this inventive disclosure) can be utilized in combination with the other inventive aspects of the invention. It should be noted that the amount ofliquid 3 contained within each of the liquid containing trough members 30 (or 30 a′ and/or 30 b′) is a function not only of the depth “d”, but also a function of the actual cross-section. Briefly, the amount ofliquid 3 present in and around the electrode(s) 1 and 5 can influence one or more effects of theadjustable plasma 4 upon theliquid 3 as well as the electrochemical interaction(s) of theelectrode 5 with theliquid 3. Further, the flow rate of the liquid 3 in and around the electrode(s) 1 and 5 can also influence many of properties of the nanocrystals formed in the resulting colloids or suspensions. These effects include not onlyadjustable plasma 4 conditioning effects (e.g., interactions of the plasma electric and magnetic fields, interactions of the electromagnetic radiation of the plasma, creation of various chemical species (e.g., Lewis acids, Bronsted-Lowry acids) within the liquid, pH changes, temperature variations of the liquid (e.g., slower liquid flow can result in higher liquid temperatures and/or longer contact or dwell time with or around theelectrodes 1/5 which can also desirably influence final products produced, such as size/shape of the formed nanocrystals, etc.) upon theliquid 3, but also the concentration or interaction of theadjustable plasma 4 with theliquid 3. Similarly, the influence of many aspects of theelectrode 5 on the liquid 3 (e.g., electrochemical interactions, temperature, etc.) is also, at least partially, a function of the amount of liquid juxtaposed to the electrode(s) 5. All of these factors can influence a balance which exists between nucleation and growth of the nanocrystals grown in theliquid 3, resulting in, for example, particle size and size range control and/or particle shape and shape range control. - Also, the initial temperature of the liquid 3 input into the trough member 30 (or 30 a′ and/or 30 b′) can also affect a variety of properties of products produced according to the disclosure herein. For example, different temperatures of the liquid 3 can affect nanocrystal size(s) and nanocrystal shape(s), concentration or amounts of various formed constituents (e.g., transient, semi-permanent or permanent constituents), ionic control of the liquid, pH, zeta potential, etc. Likewise, temperature controls along at least a portion of, or substantially all of, the trough member 30 (or 30 a′ and/or 30 b′) can have desirable effects. For example, by providing localized cooling, resultant properties of products formed (e.g., nanocrystal size(s) and/or nanocrystal shape(s)) can be controlled.
Preferable liquid 3 temperatures during the processing thereof are between freezing and boiling points, more typically, between room temperature and boiling points, and even more typically, between about 40-98 degrees C., and more typically, between about 50-98 degrees C. Such temperature can be controlled by, for example, conventional means for cooling located at or near various portions of the processing apparatus. - Further, certain processing enhancers may also be added to or mixed with the liquid(s) 3. The processing enhancers include both solids and liquids (and gases in some cases). The processing enhancer(s) may provide certain processing advantages and/or desirable final product characteristics. Some portion of the processing enhancer(s) may function, influence as or become part of, for example, desirable seed crystals (or promote desirable seed crystals, or be involved in the creation of a nucleation site) and/or crystal plane growth promoters/preventers in the electrochemical growth processes of the invention; or may simply function as a current or power regulator in the electrochemical processes of the invention. Such processing enhancers may also desirably affect current and/or voltage conditions between
electrodes 1/5 and/or 5/5. - A preferred processing enhancer is sodium bicarbonate. Examples of other process enhancers are sodium carbonate, potassium bicarbonate, potassium carbonate, trisodium phosphate, disodium phosphate, monosodium phosphate, potassium hydroxide, potassium phosphates or the like and combinations thereof. Another particularly preferred processing enhancer is a mixture of sodium bicarbonate and potassium hydroxide. Still other process enhancers to make bi-metallic nanocrystals for medical applications under certain conditions may be any material that assists in the electrochemical growth processes described herein; and any material is not substantially incorporated into or onto the surface of the gold nanocrystasl; and does not impart toxicity to the nanocrystals or to the suspension containing the nanocrystals. Processing enhancers may assist in one or more of the electrochemical reactions disclosed herein; and/or may assist in achieving one or more desirable properties in products formed according to the teachings herein. Preferably, such processing enhancers do not contain Cl− or chlorides or chlorine-based materials which are required by other processing techniques.
- For example, certain processing enhancers may dissociate into positive ions (cations) and negative ions (anions). The anions and/or cations, depending on a variety of factors including liquid composition, concentration of ions, applied fields, frequency of applied fields, waveform of the applied filed, temperature, pH, zeta potential, etc., will navigate or move toward oppositely charged electrodes. When said ions are located at or near such electrodes, the ions may take part in one or more intermediate reactions with the electrode(s) and/or other constituent(s) located at or near such electrode(s). Sometimes ions may react with one or more materials in the electrode and cause metallic ions to be produced in the liquid. Specifically, sometimes ions present in a solution between electrodes may influence material in the electrode (or near the electrode) to form metallic nano-crystals that are “grown” from material provided by the electrode. For example, certain metal ions may enter the liquid 3 from the
electrode 5 and be caused to come together (e.g., nucleate) to form constituents (e.g., ions, nanocrystals, etc.) within theliquid 3. Such ions can then be used as a raw material for the growth of bi-metallic nanocrystals. - The presence of certain nanocrystalline shapes (or shape distributions) containing specific spatially extended low index crystal planes can cause different reactions (e.g., different catalytic, electrochemical, biocatalytic and/or biophysical reactions and/or cause different biological signaling pathways to be active/inactive relative to the absence of such shaped nanoparticles) and/or different reactions selectively to occur under substantially identical conditions. Such differences in performance may be due to differing surface plasmon resonances and/or intensity of such resonances. Thus, by controlling amount (e.g., concentration), nanocrystal sizes, the presence or absence of certain extended growth crystal planes, and/or nanocrystalline shapes or shape distribution(s), certain reactions (e.g., catalytic, electrochemical, biological reactions and/or biological signaling pathways) can be desirably influenced and/or controlled. Such control can result in the prevention and/or treatment of a variety of different diseases or indications that are a function of certain biologic reactions and/or signaling pathways, as well as control of a number of non-biological reaction pathways.
- Further, certain processing enhancers may also include materials that may function as charge carriers, but may themselves not be ions. Specifically, metallic-based particles, either introduced or formed in situ (e.g., heterogeneous or homogenous nucleation/growth) by the electrochemical processing techniques disclosed herein, can also function as charge carriers, crystal nucleators and/or growth promoters, which may result in the formation of a variety of different crystalline shapes (e.g., hexagonal plates, octahedrons, techahedrons, pentagonal bi-pyramids (decahedrons), etc.). Once again, the presence of particular particle crystal sizes, extended crystal planes and/or shapes or shape distributions of such crystals, can desirably influence certain reactions (e.g., binding to a particular protein or protein homologue and/or affecting a particular biological signaling pathway such as an inflammatory pathway or a proteasomal pathway) to occur.
- For example, in reference to
FIGS. 9 and 10 a-10 d, platinum species that are formed in afirst trough member 30 a′/30 b′ are caused to flow into asecond trough member 30 a′/30 b′ and take part in the formation of bi-metallic nanocrystals therein. More specifically, a first set of electrochemical reactions occur in a water containing a suitable processing enhancer to create a modified water-processing enhancer solution/suspension, which then serves as a raw material supply for a second set of electrochemical reactions that occur in asecond trough member 30 a′/30 b′. In some cases, the two separate trough members are kept as separate members and the output of the first trough member is allowed to cool before being input into the second trough member. However, in another embodiment, the two trough members can be an integral unit, with or without cooling means located between the twoidentifiable portions 30 a′/30 b′. - Further, since the processing enhancers of the present invention do not contemplate those traditional organic-based molecules used in traditional reduction chemistry techniques, the lack of such chemical reductant (or added surfactant) means that the surfaces of the grown nanocrystals on the invention are very “clean” relative to nanoparticles that are formed by traditional reduction chemistry approaches. It should be understood that when the term “clean” is used with regard to nanocrystal surfaces or when the phrase “substantially free from organic impurities or films” (or a similar phrase) is used, what is meant is that the formed nanocrystals do not have chemical constituents adhered or attached to their surfaces which (1) alter the functioning of the nanocrystal and/or (2) form a layer, surface or film which covers a significant portion (e.g., at least 25% of the crystal, or more typically, at least 50% of the crystal). In preferred embodiments, the nanocrystal surfaces are completely free of any organic contaminants or reactants which materially change their functionality. It should be further understood that incidental components that are caused to adhere to nanocrystals of the invention and do not adversely or materially affect the functioning of the inventive nanocrystals, should still be considered to be within the metes and bounds of the invention.
- The lack of added chemicals (e.g., organics or chlorine-based materials) permits the growth of the metal atoms and also does not adversely affect the performance of the nanocrystals (e.g., in catalysis reactions or in biological reactions, in vivo it affects the protein corona formed around the nanoparticles/nanocrystals in, for example, serum and/or reduces toxic compounds introduced into cells or or an organism). For example, but without wishing to be bound by any particular theory or explanation, in biological reactions, protein corona formation can control location of a nanoparticle/nanocrystal in vivo, as well as control protein folding of proteins at or near the nanoparticle/nanocrystal surfaces. Such differences in performance may be due to such factors including, but not limited to, surface charge, surface plasmon resonance, epitaxial effects, surface double layers, zones of influence, toxic surface contaminents and others. Such novel shapes also affect, for example, catalysis.
- Still further, once a seed crystal occurs in the process and/or a set of extended crystal planes begins to grow (e.g., homogenous nucleation) or a seed crystal is separately provided (e.g., heterogenous nucleation) the amount of time that a formed particle (e.g., a metal atom) is permitted to dwell at or near one or more electrodes in an electrochemical process can result in the size of bi-metallic nanocrystals increasing as a function of time (e.g., metal atoms can assemble into metal nanocrystals and, if unimpeded by certain organic constituents in the liquid, they can grow into a variety of shapes and sizes). The amount of time that crystal nucleation/growth conditions are present can control the shape(s) and sizes(s) of grown bi-metallic nanocrystals. Accordingly, dwell time at/around electrodes, liquid flow rate(s), trough cross-sectional shape(s), etc, all contribute to nanocrystal growth conditions, as discussed elsewhere herein.
- In many of the preferred embodiments herein, one or more AC sources are utilized (e.g., transformer(s) 60 and power supply 501AC). The rate of change from “+” polarity on one electrode to “−” polarity on the same electrode is known as Hertz, Hz, frequency, or cycles per second. In the United States, the standard output frequency is 60 Hz, while in Europe it is predominantly 50 Hz. As shown in the Examples herein, the frequency can also influence size and/or shape and/or presence of nanocrystals and/or ions formed according to the electrochemical techniques disclosed herein. Preferable frequencies are 5-1000 Hz, more typically, 20-500 Hz, even more typically, 40-200 Hz, and even more typically, 50-100 Hz. For example, and without wishing to be bound by any particular theory or explanation, nucleated or growing crystals can first have attractive forces exerted on them (or on crystal growth constituents, such as ions or atoms, taking part in forming the crystal(s)) due to, for example, unlike charges attracting and then repulsive forces being exerted on such constituents (e.g., due to like charges repelling). These factors also clearly play a large role in nucleation and/or crystal growth of the novel nanocrystals formed by affecting particle size and/or shapes; as well as permitting the crystals to be formed without the need for reductants or surfactants (i.e., that needed to be added to take part in the prior art reduction chemistry techniques) causing the nanocrystal surfaces to be free of such added chemical species. The lack of organic-based coatings on the surface of grown nanocrystals alters (and in some cases controls) their biological function. Further, when water is used as the liquid, hydrolysis can occur at the electrodes, resulting in gas production and the production of other lysis products of water including hydrated electrons, OH−, H*, H3O, H2O2, etc. Such lysis products also may assist in the crystal growth processes disclosed herein and/or assist in the stabilization of the bi-metallic nanocrystals in the suspension.
- Moreover, the particular waveform that is used for a specific frequency also affects nanocrystal growth conditions, and thus effects nanocrystal size(s) and/or shape(s). While the U.S. uses a standard AC frequency of 60 Hz, it also uses a standard waveform of a “sine” wave. As shown in the Examples herein, changing the waveform from a sine wave to a square wave or a triangular wave also affects nanocrystal crystallization conditions and thus affects resultant nanocrystal size(s) and shape(s). Preferred waveforms include sine waves, square waves and triangular waves, however hybrid waveforms should be considered to be within the metes and bounds of the invention.
- Still further, the voltage applied in the novel electrochemical techniques disclosed herein can also affect nanocrystalline size(s) and shape(s). A preferred voltage range is 20-2000 Volts, a more preferred voltage range is 50-1000 Volts and an even more preferred voltage range is 100-300 Volts. In addition to voltage, the amperages used with these voltages typically are 0.1-10 Amps, a more preferred amperage range is 0.1-5 Amps and an even more preferred amperage range is 0.4-1 Amps per electrode set under the processing parameters disclosed herein.
- Still further, the “duty cycle” used for each waveform applied in the novel electrochemical techniques disclosed herein can also affect nanocrystalline size(s) and shape(s). In this regard, without wishing to be bound by any particular theory or explanation, the amount of time that an electrode is positively biased can result in a first set of reactions, while a different set of reactions can occur when the electrode is negatively biased. By adjusting the amount of time that the electrodes are positively or negatively biased, size(s) and/or shape(s) of grown nanocrystals can be controlled. Further, the rate at which an electrode converts to + or − is also a function of waveform shape and also influences nanocrystal size(s) and/or shape(s).
- Temperature can also play an important role. In some of the preferred embodiments disclosed herein, the boiling point temperature of the water is approached in at least a portion of the processing vessel where nanocrystals are nucleated and grown. For example, output water temperature in the continuous processing Examples herein ranges from about 60° C.-99° C. However, as discussed elsewhere herein, different temperature ranges are also desirable. Temperature can influence resultant product (e.g., size and/or shape of nanocrystals) as well as the amount of resultant product (i.e., ppm level of nanocrystals in the suspension or colloid). For example, while it is possible to cool the liquid 3 in the
trough member 30 by a variety of known techniques (as disclosed in some of the Examples herein), many of the Examples herein do not cool the liquid 3, resulting in evaporation of a portion of the liquid 3 during processing thereof. - It should be understood that a variety of different shapes and/or cross-sections can exist for the trough member 30 (or 30 a′ and/or 30 b′), any one of which can produce desirable results as a function of a variety of design and production considerations. For example, one or more constituents produced in the portion(s) 30 a′, or 30 b′ could be transient (e.g., a seed crystal or nucleation point) and/or semi permanent (e.g., grown nanocrystals present in a colloid). If such constituent(s) produced, for example, in
portion 30 a′ is to be desirably and controllably reacted with one or more constituents produced in, for example,portion 30 b′, then a final product (e.g., properties of a final product) which results from such mixing could be a function of when constituents formed in theportions 30 a′ and 30 b′ are mixed together. Further, transient constituents formed in afirst trough member 30 a′/30 b′ can also affect subsequent bi-metallic nanocrystal formation in asecond trough member 30 a′/30 b′. Thus, the amount of time that lapses between the production of a first aqueous product in a first trough member and wherein such first product becomes a raw material in a second trough member can also influence the bi-metallic nanocrystal suspension formed. Thus, the temperature of liquids entering and exiting can be monitored/controlled to maximize certain desirable processing conditions and/or desirable properties of final products and/or minimize certain undesirable products. Still further, processing enhancers may be selectively utilized in one or more of the portions of the different trough members. -
FIG. 6 shows a schematic view of the general apparatus utilized in accordance with the teachings of some of the preferred embodiments of the present invention. In particular, thisFIG. 6 shows a side schematic view of thetrough member 30 containing a liquid 3 therein. On the top of thetrough member 30 rests a plurality ofcontrol devices 20 a-20 d which are, in this embodiment, removably attached thereto. Thecontrol devices 20 a-20 d may of course be permanently fixed in position when practicing various embodiments of the invention. The precise number of control devices 20 (and corresponding electrode(s) 1 and/or 5 as well as the configuration(s) of such electrodes) and the positioning or location of the control devices 20 (andcorresponding electrodes 1 and/or 5) are a function of various preferred embodiments of the invention discussed in greater detail elsewhere herein. However, in general, an input liquid 3 (for example water or purified water containing a process enhancer) is provided to a liquid transport means 40 (e.g., a liquid pump, gravity or liquid pumping means for pumping the liquid 3) such as aperistaltic pump 40 for pumping theliquid 3 into thetrough member 30 at a first-end 31 thereof. The liquid transport means 40 may include any means for movingliquids 3 including, but not limited to a gravity-fed or hydrostatic means, a pumping means, a regulating or valve means, etc. However, the liquid transport means 40 should be capable of reliably and/or controllably introducing known amounts of the liquid 3 into thetrough member 30. The amount of time that theliquid 3 is contained within the trough member 30 (e.g., at or around one or more electrode(s) 1/5) also influences the products produced (e.g., the sizes(s) and/or shapes(s) of the grown nanocrystals). - Once the
liquid 3 is provided into thetrough member 30, means for continually moving theliquid 3 within thetrough member 30 may or may not be required. However, a simple means for continually moving theliquid 3 includes thetrough member 30 being situated on a slight angle θ (e.g., less than a degree to a few degrees for alow viscosity fluid 3 such as water) relative to the support surface upon which thetrough member 30 is located. For example, a difference in vertical height of less than one inch between aninlet portion 31 and anoutlet portion 32, spaced apart by about 6 feet (about 1.8 meters) relative to the support surface may be all that is required, so long as the viscosity of theliquid 3 is not too high (e.g., any viscosity around the viscosity of water can be controlled by gravity flow once such fluids are contained or located within the trough member 30). The need for a greater angle θ could be a result of processing aliquid 3 having a viscosity higher than water; the need for the liquid 3 to transit thetrough 30 at a faster rate, etc. Further, when viscosities of the liquid 3 increase such that gravity alone is insufficient, other phenomena such as specific uses of hydrostatic head pressure or hydrostatic pressure can also be utilized to achieve desirable fluid flow. Further, additional means for moving theliquid 3 along thetrough member 30 could also be provided inside thetrough member 30. Such means for moving the fluid include mechanical means such as paddles, fans, propellers, augers, etc., acoustic means such as transducers, thermal means such as heaters and/or chillers (which may have additional processing benefits), etc., are also desirable for use with the present invention. -
FIG. 6 also shows a storage tank orstorage vessel 41 at theend 32 of thetrough member 30.Such storage vessel 41 can be any acceptable vessel and/or pumping means made of one or more materials which, for example, do not negatively interact with the liquid 3 (or constituents contained therein) produced within thetrough member 30. Acceptable materials include, but are not limited to plastics such as high density polyethylene (HDPE), glass, metal(s) (such a certain grades of stainless steel), etc. Moreover, while astorage tank 41 is shown in this embodiment, thetank 41 should be understood as including a means for distributing or directly bottling or packaging thefluid 3 processed in thetrough member 30. - The electrode control devices shown generally in, for example,
FIGS. 2 and 6 are shown in greater detail inFIG. 8 c . In particular,FIG. 8 c shows a perspective view of thecontrol device 20.FIG. 8 c shows abase portion 25 is provided, said base portion having atop portion 25′ and abottom portion 25″. Thebase portion 25 is made of a suitable rigid plastic material including, but not limited to, materials made from structural plastics, resins, polyurethane, polypropylene, nylon, teflon, polyvinyl, etc. A dividingwall 27 is provided between two electrode adjustment assemblies. The dividingwall 27 can be made of similar or different material from that material comprising thebase portion 25. Two servo-step motors 21 a and 21 b are fixed to thesurface 25′ of thebase portion 25. Thestep motors 21 a, 21 b could be any step motor capable of slightly moving (e.g., on a 360 degree basis, slightly less than or slightly more than 1 degree) such that a circumferential movement of the step motors 21 a/21 b results in a vertical raising or lowering of anelectrode component 23 a is the drivewheel connected to the output shaft 231 a of the drive motor 21 a such that when the drive shaft 231 a rotates, circumferential movement of thewheel 23 a is created. Further, aslave wheel 24 a is caused to press against and toward thedrivewheel 23 a such that frictional contact exists therebetween. Thedrivewheel 23 a and/orslavewheel 24 a may include a notch or groove on an outer portion thereof to assist in accommodating theelectrodes drivewheel 23 a by a spring 285 located between the portions 241 a and 261 a attached to theslave wheel 24 a. In particular, a coiled spring 285 can be located around the portion of the axis 262 a that extends out from the block 261 a. Springs should be of sufficient tension so as to result in a reasonable frictional force between the drivewheel 24 a and the slavewheel 24 a such that when the shaft 231 a rotates a determined amount, theelectrode assemblies base portion 25. Such rotational or circumferential movement of the drivewheel 23 a results in a direct transfer of vertical directional changes in theelectrodes - The drive motors 21 a/21 b can be any suitable drive motor which is capable of small rotations (e.g., slightly below 1°/360° or slightly above 1°/360°) such that small rotational changes in the drive shaft 231 a are translated into small vertical changes in the electrode assemblies. A preferred drive motor includes a drive motor manufactured by RMS Technologies model 1MC17-S04 step motor, which is a DC-powered step motor. This step motors 21 a/21 b include an RS-232
connection 22 a/22 b, respectively, which permits the step motors to be driven by a remote control apparatus such as a computer or a controller. - The
portions base portion 25 relative to thetrough member 30. Theportions base portion 25. The portions 274 a/274 b and 275 a/275 b can also be made of the same, similar or different material from thebase portion 25. However, these portions should be electrically insulating in that they house various wire components associated with delivering voltage and current to theelectrode assemblies 1 a/1 b, 5 a/5 b, etc. - With regard to the size of the
control device 20 shown inFIG. 8 c , length and width can be any dimension which accommodates the size of the step motors 21 a/21 b, and the width of thetrough member 30. In this regard, length should be at least as long as thetrough member 30 is wide, and typically slightly longer (e.g., 10-30%). The width needs to be wide enough to house the step motors 21 a/21 b and not be so wide as to unnecessarily underutilize longitudinal space along the length of thetrough member 30. In one preferred embodiment of the invention, the length is about 7 inches (about 19 millimeters) and the width is about 4 inches (about 10.5 millimeters). The thickness of thebase member 25 is any thickness sufficient which provides structural, electrical and mechanical rigidity for thebase member 25 and should be of the order of about ¾″-¾″ (about 6 mm-19 mm). While these dimensions are not critical, the dimensions give an understanding of size generally of certain components of one preferred embodiment of the invention. - Further, the base member 25 (and the components mounted thereto), can be covered by a suitable cover (not shown) to insulate electrically, as well as creating a local protective environment for all of the components attached to the
base member 25. Such cover can be made of any suitable material which provides appropriate safety and operational flexibility. Exemplary materials include plastics similar to that used for other portions of thetrough member 30 and/or thecontrol device 20 and are typically transparent. This cover member can also be made of the same type of materials used to make thebase portion 25. The cover can include through-holes which can be aligned with excess portions of, for example,electrodes 5, which can be connected to, for example, a spool of electrode wire (not shown in these drawings). - As shown in
FIG. 8 j , theportions wire power sources 60 or 501AC and theelectrodes 1/5. The servo-motor 21 a functions as discussed above, but two electrodes are driven by a single servo drive motor 21 a. Accordingly, a single drive motor 21 a can replace two drive motors in the case of the embodiment shown inFIG. 8 j . Further, by providing the electrical contact between thewires 1/5 and thepower sources 60/501AC, all electrical connections are provided on a top surface of (i.e., the surface further away from the liquid 3) resulting in certain design and production advantages. -
FIG. 8 c shows arefractory material component 29 a, 29 b. Thecomponent 29 is made of, for example, suitable refractory component, including, for example, aluminum oxide or the like. Therefractory component 29 may have a transverse through-hole therein which provides for electrical connections to the electrode(s) 1 and/or 5. Further a longitudinal through-hole is present along the length of therefractory component 29 such thatelectrode assemblies 1/5 can extend therethrough. -
FIG. 8 c specifically shows one electrode(s) 1 a as extending through a first refractory portion 29 a and one electrode(s) 5 a is shown as extending through a secondrefractory portion 29 b. Accordingly, each of the electrode assemblies expressly disclosed herein, as well as those referred to herein, can be utilized in combination with the preferred embodiments of the control device shown herein. - In order for the
control devices 20 to be actuated, two general processes need to occur. A first process involves electrically activating the electrode(s) 1 and/or 5 (e.g., applying power thereto from a preferred power source 10), and the second general process occurrence involves determining, for example, how much power (e.g., voltage and/or current) is applied to the electrode(s) and appropriately adjustingelectrode 1/5 height in response to such determinations (e.g., manually and/or automatically adjusting the height of theelectrodes 1/5); or adjusting the electrode height or simply moving the electrode into (e.g., progressively advancing the electrode(s) 5 through the liquid 3) or out of contact with theliquid 3, as a function of time. In the case of utilizing acontrol device 20, suitable instructions are communicated to thestep motor 21 through the RS-232ports control device 20, as well as the electrode activation process, are discussed herein. - A preferred embodiment of the invention utilizes the
automatic control devices 20 shown in various figures herein. Thestep motors 21 a and 21 b shown in, for example,FIG. 8 c . Theelectrodes 1/5 are monitored either by the electrical circuit diagrammed in each ofFIGS. 8 d-8 h (e.g., for electrode sets 1/5 that make aplasma 4 or for electrode sets 5/5); or are monitored by the electrical circuit diagrammed in each ofFIGS. 8 g and 8 i for electrode sets 5/5, in some embodiments herein. - In particular, in this embodiment, the electrical circuit of
FIG. 8 h is a voltage monitoring circuit. Specifically, voltage output from each of the output legs of thesecondary coil 603 in thetransformer 60 are monitored over the points “P-Q” and the points “P′-Q′”. Specifically, the resistor denoted by “RL” corresponds to the internal resistance of the multi-meter measuring device (not shown). The output voltages measured between the points “P-Q” and “P′-Q′” typically, for several preferred embodiments shown in the Examples later herein, range between about 200 volts and about 4,500 volts. However, higher and lower voltages can work with many of the embodiments disclosed herein. Desirable target voltages have been determined for each electrode set 1 and/or 5 at each position along atrough member 30 a′. Such desirable target voltages are achieved as actual applied voltages by, utilizing, for example, the circuit control shown inFIGS. 8 d, 8 e and 8 f . TheseFIGS. 8 d, 8 e and 8 f refer to sets of relays controlled by a Velleman K8056 circuit assembly (having a micro-chip PIC16F630-I/P). Eachtransformer 60 is connected electrically in a manner shown inFIG. 8 h . Eachtransformer 60 and associated measuring points “P-Q” and “P′-Q′” are connected to an individual relay. For example, the points “P-Q” correspond to relaynumber 501 inFIG. 8 d and the points “P′-Q”′ correspond to therelay 502 inFIG. 8 d . Accordingly, two relays are required for eachtransformer 60. Each relay, 501, 502, etc., sequentially interrogates a first output voltage from a first leg of asecondary coil 603 and then a second output voltage from a second leg of thesecondary coil 603; and such interrogation continues onto a first output voltage from a second transformer 60 b on a first leg of itssecondary coil 603, and then on to a second leg of thesecondary coil 603, and so on. - The computer or logic control for the disclosed interrogation voltage adjustment techniques are achieved by any conventional program or controller, including, for example, in a preferred embodiment, standard visual basic programming steps utilized in a PC. Such programming steps include interrogating, reading, comparing, and sending an appropriate actuation symbol (e.g., raise or lower an electrode relative to the
surface 2 of the liquid 3). Such techniques should be understood by an artisan of ordinary skill. - Further, in another preferred embodiment of the invention utilized in Example 1 for the electrode sets 5/5′, the
automatic control devices 20 are controlled by the electrical circuits ofFIGS. 8 e, 8 f, 8 g and 8 i . In particular, the electrical circuit ofFIG. 8 i is a voltage monitoring circuit used to measure current. In this case, voltage and current are the same numerical value due to choice of a resistor (discussed later herein). Specifically, voltage output from each power source 501AC is monitored over the points “P-Q” and the points “P′-Q′”. Specifically, the resistor denoted by “RL” corresponds to the internal resistance of the multi-meter measuring device (not shown). The output voltages measured between the points “P-Q” and “P′-Q”′ typically, for several preferred embodiments shown in the Examples later herein, range between about 0.05 volts and about 5 volts. However, higher and lower voltages can work with many of the embodiments disclosed herein. Desirable target voltages have been determined for each electrode set 5/5′ at each position along atrough member 30 b′. Such desirable target voltages are achieved as actual applied voltages by, utilizing, for example, the circuit control shown inFIGS. 8 e, 8 f, 8 g and 8 i . TheseFIG. 8 refer to sets of relays controlled by a Velleman K8056 circuit assembly (having a micro-chip PIC16F630-I/P). - In particular, the servo-
motor 21 is caused to rotate at a specific predetermined time in order to maintain adesirable electrode 5 profile. The servo-motor 21 responds by rotating a predetermined amount in a clockwise direction. Specifically the servo-motor 21 rotates a sufficient amount such that about 0.009 inches (0.229 mm) of theelectrode 5 is advanced toward and into the female receiver portion o5 (shown, for example inFIGS. 10 b and 11 a ). Thus, theelectrode 5 is progressively advanced through theliquid 3. In one preferred embodiment discussed herein,such electrode 5 movement occurs about every 4.3 minutes. Accordingly, the rate of vertical movement of eachelectrode 5 into the female receiver portion o5 is about 1 inch (about 1.9 cm) every 8 hours. Accordingly, a substantiallyconstant electrode 5 shape or profile is maintained by its constant or progressive advance into and through theliquid 3. Further, once the advancing end of theelectrode 5 reaches the longitudinal end of the female receiver portion o5, theelectrode 5 can be removed from the processing apparatus. Alternatively, an electrode collecting means for collecting the “used” portion of the electrode can be provided. - Such means for collecting the electrode(s) 5 include, but are not limited to, a winding or spooling device, and extended portion o5, a wire clipping or cutting device, etc. However, in order to achieve different current/voltage profiles (and thus a variety of different nanocrystal size(s) and/or shapes(s), other rates of electrode movement are also within the metes and bounds of this invention.
- Moreover, with specific reference to
FIGS. 8 e, 8 f, 8 g and 8 i , it should be noted that an interrogation procedure occurs sequentially by determining the voltage of each electrode, which in the embodiments herein, are equivalent to the amps because inFIG. 8 i the resistors Ra and Rb are approximately 1 ohm, accordingly, V=I. In other words, each power source 501AC is connected electrically in a manner shown inFIGS. 8 e, 8 f, 8 g and 8 i . Each power source 501AC and associated measuring points “P-Q” and “P′-Q′” are connected to two individual relays. For example, the points “P-Q” correspond to relaynumber FIG. 8 g and the points “P′-Q′” correspond to therelay FIG. 8 g . Accordingly, relays are required for each electrode set 5/5. Each relay, 501/501′ and 502/502′, etc., sequentially interrogates the output voltage from the power source 501AC and then a second voltage from the same power source 501AC, and so on. - The computer or logic control for the disclosed electrode height adjustment techniques are achieved by any conventional program or controller, including, for example, in a preferred embodiment, standard visual basic programming steps utilized in a PC. Such programming steps include reading and sending an appropriate actuation symbol to lower an electrode relative to the
surface 2 of theliquid 3. Such techniques should be understood by an artisan of ordinary skill. - For purposes of the present invention, the terms and expressions below, appearing in the Specification and Claims, are intended to have the following meanings:
- “Substantially clean”, as used herein should be understood when used to describe nanocrystal surfaces means that the nanocrystals do not have chemical constituents adhered or attached to their surfaces in such an amount that would materially alter the functioning of the nanocrystal in at least one of its significant properties of the metallic-based nanocrystals set forth in the Examples herein. Alternatively, the metallic-based nanocrystal does not have a layer, surface or film which covers a significant portion (e.g., at least 25% of the crystal, or in another embodiment at least 50% of the crystal). It also can mean that the nanocrystal surfaces are completely free of any organic contaminants which materially change their functionality over bare gold crystal surfaces. It should be understood that incidental components that are caused to adhere to nanocrystals of the invention and do not adversely or materially affect the functioning of the inventive nanocrystals, should still be considered to be within the metes and bounds of the invention. The term should also be understood to be a relative term referencing the lack of traditional organic-based molecules (i.e., those used in traditional reduction chemistry techniques) on the surfaces of the grown nanocrystals of the invention.
- As used herein, the term “processing-enhancer” or “processing-enhanced” or “process enhancer” means at least one material (e.g., solid, liquid and/or gas) and typically means an inorganic material, which material does not significantly bind to the formed nanocrystals, but rather facilitates nucleation/growth during an electrochemical-stimulated growth process. The material serves important roles in the process including providing charged ions in the electrochemical solution to permit the crystals to be grown. The process enhancer is critically a compound(s) which remains in solution, and/or does not form a coating (in one embodiment an organic coating), and/or does not adversely affect the formed nanocrystals or the formed suspension(s), and/or is destroyed, evaporated, or is otherwise lost during the electrochemical crystal growth process.
- The phrase “trough member” as used herein should be understood as meaning a large variety of fluid handling devices including, pipes, half pipes, channels or grooves existing in materials or objects, conduits, ducts, tubes, chutes, hoses and/or spouts, so long as such are compatible with the electrochemical processes disclosed herein.
- The following Examples serve to illustrate certain embodiments of the invention but should not to be construed as limiting the scope of the disclosure as defined in the appended claims.
- In general, this Example utilizes certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c, and 11 a. Alltrough members 30 a′ and 30 b′ in the aforementioned Figures were made from ⅛″ (about 3 mm) thick plexiglass, and ¼″ (about 6 mm) thick polycarbonate, respectively. The support structure 34 (not shown in many of the Figures but shown inFIG. 9 ) was also made from plexiglass which was about ¾″ thick (about 6-7 mm thick). Eachtrough member 30 a′ was integral withtrough member 30 b′. The cross-sectional shape of thetrough member 30 a′ used in this Example corresponded to that shape shown inFIG. 5 b (i.e., was a trapezoidal-shaped cross-section). Relevant dimensions for 30 a′ were “S,S′” which measured about 1.5″ (about 3.81 cm), “M” which measured about 2.5″ (about 6.35 cm), “R” measured about ¾″ (about 1.9 cm) and “d′” which measured about ½″ (about 1.3 cm). - Each
trough member portion 30 b′ had a cross-sectional shape corresponding toFIG. 5 a . The relevant dimensions fortrough member portion 30 b′ are reported in Table 1 as “M” (i.e., inside width of the trough at the entrance and exact portion of thetrough member 30 b′), “LT” (i.e., transverse length or flow length of thetrough member 30 b′), “S” (i.e., the height of thetrough member 30 b′), and “d″” (i.e., depth of the liquid 3″ within thetrough member 30 b′). The thickness of each sidewall portion oftrough 30 b′ also measured about ¼″ (about 6 mm) thick. - The
water 3 used in Example 1 as an input into thetrough member 30 a′ (and used in Examples 1-17 in combination with a processing enhancer) was produced by a Reverse Osmosis process and deionization process (referred to herein as de-ionized water). In essence, Reverse Osmosis (RO) is a pressure driven membrane separation process that separates species that are dissolved and/or suspended substances from the ground water. It is called “reverse” osmosis because pressure is applied to reverse the natural flow of osmosis (which seeks to balance the concentration of materials on both sides of the membrane). The applied pressure forces the water through the membrane leaving the contaminants on one side of the membrane and the purified water on the other. The reverse osmosis membrane utilized several thin layers or sheets of film that are bonded together and rolled in a spiral configuration around a plastic tube. (This is also known as a thin film composite or TFC membrane.) In addition to the removal of dissolved species, the RO membrane also separates out suspended materials including microorganisms that may be present in the water. After RO processing a mixed bed deionization filter was used. The total dissolved solvents (“TDS”) after both treatments was about 0.2 ppm, as measured by an Accumet® AR20 pH/conductivity meter. -
TABLE 1 Run ID: NE10214 Flow In (ml/min) 230 Rate: Out (ml/min) 220 Volts: Set # 1750 Set #'s 2-8 220 Set #'s 2-8 60 frequency, Hz PE/Concentration (mg/mL) 0.528 Wire Diameter (mm) 1.0 Contact “WL” (in/mm) 1/25.4 Electrode Separation “y” (in/mm) .25/6.4 Electrode Config. Figure 8b, 11a Produced Au PPM 6.6 Output Temp ° C. at 32 72 Dimensions Plasma 4 Figs. 9 Process Figures 10c M (in/mm) 1.5/38 L T (in/mm)36/914 d″ (in/mm) 1/25 S (in/mm) 1.5/38 Electrode Curr. (A) 0.71 Total Curr. Draw (A) 5 Hydrodynamic r (nm) 19.43 TEM Avg. Dia. (nm) 12.38 “c-c” (mm) 76 Set 1electrode # 1a “x” (in/mm) 0.25/6.4 electrode # 5a “c-c” (mm) 102 Set 2electrode # 5b “x” (in/mm) n/a electrode # 5b′ “c-c” (mm) 76 Set 3electrode # 5c electrode # 5c′ “c-c” (mm) 76 Set 4electrode # 5d electrode # 5d′ “c-c” (mm) 127 Set 5electrode # 5e electrode # 5e′ “c-c” (mm) 127 Set 6electrode # 5f electrode # 5f′ “c-c” (mm) 152 Set 7electrode # 5g electrode # 5g′ “c-c” (mm) 178 Set 8electrode # 5h electrode # 5h′ “c-c” (mm) 76 - Table 1 shows that the amount of processing enhancer (PE) (NaHCO3) that was added to purified water was about 0.53 mg/ml. It should be understood that other amounts of this processing enhancer also function within the metes and bounds of the invention. The purified water/NaHCO3 mixture was used as the
liquid 3 input intotrough member 30 a′. The depth “d′” of the liquid 3′ in thetrough member 30 a′ (i.e., where the plasma(s) 4 is formed) was about 7/16″ to about ½″ (about 11 mm to about 13 mm) at various points along thetrough member 30 a′. The depth “d′” was partially controlled through use of the dam 80 (shown inFIG. 9 ). Specifically, thedam 80 was provided near theoutput end 32 of thetrough member 30 a′ and assisted in creating the depth “d′” (shown inFIG. 5 b as “d”) to be about 7/6″-½″ (about 11-13 mm) in depth. The height of thedam 80 measured about ¾″ (about 6 mm) and the longitudinal length measured about ½″ (about 13 mm). The width was completely across the bottom dimension “R” of thetrough member 30 a′. Accordingly, the total volume ofliquid 3′ in thetrough member 30 a′ during operation thereof was about 2.14 in3 (about 35 ml) to about 0.89 in3 (about 14.58 ml). - The rate of flow of the liquid 3′ into the
trough member 30 a′ as well as intotrough member 30 b′, was about 230 ml/minute and the rate of flow out of thetrough member 30 b′ at thepoint 32 was about 220 ml/minute (i.e., due to evaporation). Other acceptable flow rates should be considered to be within the metes and bounds of the invention. - Such flow of
liquid 3′ was obtained by utilizing a Masterflex® L/S pump drive 40 rated at 0.1 horsepower, 10-600 rpm. The model number of theMasterflex® pump 40 was 7523-80. The pump drive had a pump head also made by Masterflex® known as Easy-Load Model No. 77201-60. In general terms, the head for thepump 40 is known as a peristaltic head. The precise settings on the pump was 230 milliliters per minute. Tygon® tubing having a diameter of ¼″ (i.e., size 06419-25) was placed into the peristaltic head. The tubing was made by Saint Gobain for Masterflex®. One end of the tubing was delivered to afirst end 31 of thetrough member 30′a. - Table 1 shows that there was a single electrode set 1 a/5 a. The power source for each electrode set 1/5 was an
AC transformer 60. Specifically,FIG. 7 a shows a source ofAC power 62 connected to atransformer 60. In addition, acapacitor 61 is provided so that, for example, loss factors in the circuit can be adjusted. The output of thetransformer 60 is connected to the electrode(s) 1/5 through thecontrol device 20. A preferred transformer for use with the present invention is one that uses alternating current flowing in aprimary coil 601 to establish an alternating magnetic flux in acore 602 that easily conducts the flux. - When a
secondary coil 603 is positioned near theprimary coil 601 andcore 602, this flux will link thesecondary coil 603 with theprimary coil 601. This linking of thesecondary coil 603 induces a voltage across the secondary terminals. The magnitude of the voltage at the secondary terminals is related directly to the ratio of the secondary coil turns to the primary coil turns. More turns on thesecondary coil 603 than theprimary coil 601 results in a step up in voltage, while fewer turns results in a step down in voltage. - Preferred transformer(s) 60 for use in these Examples have deliberately poor output voltage regulation made possible by the use of magnetic shunts in the
transformer 60. Thesetransformers 60 are known as neon sign transformers. This configuration limits current flow into the electrode(s) 1/5. With a large change in output load voltage, thetransformer 60 maintains output load current within a relatively narrow range. - The
transformer 60 is rated for its secondary open circuit voltage and secondary short circuit current. Open circuit voltage (OCV) appears at the output terminals of thetransformer 60 only when no electrical connection is present. Likewise, short circuit current is only drawn from the output terminals if a short is placed across those terminals (in which case the output voltage equals zero). However, when a load is connected across these same terminals, the output voltage of thetransformer 60 should fall somewhere between zero and the rated OCV. In fact, if thetransformer 60 is loaded properly, that voltage will be about half the rated OCV. - The
transformer 60 is known as a Balanced Mid-Point Referenced Design (e.g., also formerly known as balanced midpoint grounded). This is most commonly found in mid to higher voltage rated transformers and most 60 mA transformers. This is the only type transformer acceptable in a “mid-point return wired” system. The “balanced”transformer 60 has oneprimary coil 601 with twosecondary coils 603, one on each side of the primary coil 601 (as shown generally in the schematic view inFIG. 7 bg). Thistransformer 60 can in many ways perform like two transformers. Just as the unbalanced midpoint referenced core and coil, one end of eachsecondary coil 603 is attached to thecore 602 and subsequently to the transformer enclosure and the other end of the eachsecondary coil 603 is attached to an output lead or terminal. Thus, with no connector present, an unloaded 15,000 volt transformer of this type, will measure about 7,500 volts from each secondary terminal to the transformer enclosure but will measure about 15,000 volts between the two output terminals. - In alternating current (AC) circuits possessing a line power factor of 1 (or 100%), the voltage and current each start at zero, rise to a crest, fall to zero, go to a negative crest and back up to zero. This completes one cycle of a typical sine wave. This happens 60 times per second in a typical US application. Thus, such a voltage or current has a characteristic “frequency” of 60 cycles per second (or 60 Hertz) power. Power factor relates to the position of the voltage waveform relative to the current waveform. When both waveforms pass through zero together and their crests are together, they are in phase and the power factor is 1, or 100%.
FIG. 7 c shows two waveforms “V” (voltage) and “C” (current) that are in phase with each other and have a power factor of 1 or 100%; whereasFIG. 7 d shows two waveforms “V” (voltage) and “C” (current) that are out of phase with each other and have a power factor of about 60%; both waveforms do not pass through zero at the same time, etc. The waveforms are out of phase and their power factor is less than 100%. - The normal power factor of most
such transformers 60 is largely due to the effect of themagnetic shunts 604 and thesecondary coil 603, which effectively add an inductor into the output of the transformer's 60 circuit to limit current to theelectrodes 1/5. The power factor can be increased to a higher power factor by the use of capacitor(s) 61 placed across theprimary coil 601 of the transformer, 60 which brings the input voltage and current waves more into phase. - The unloaded voltage of any
transformer 60 to be used in the present invention is important, as well as the internal structure thereof. Desirable unloaded transformers for use in the present invention include those that are around 9,000 volts, 10,000 volts, 12,000 volts and 15,000 volts. However, these particular unloaded volt transformer measurements should not be viewed as limiting the scope acceptable power sources as additional embodiments. A specific desirable transformer for use in these Examples is made by Franceformer, Catalog No. 9060-P-E which operates at: primarily 120 volts, 60 Hz; and secondary 9,000 volts, 60 mA. - Accordingly, each
transformer assembly 60 a-60 h (and/or 60 a′-60 h′; and/or 60 a″-60 h″) can be the same transformer, or can be a combination of different transformers (as well as different polarities). The choice of transformer, power factor, capacitor(s) 61, polarity, electrode designs, electrode location, electrode composition, cross-sectional shape(s) of thetrough member 30 a′, local or global electrode composition, atmosphere(s), local orglobal liquid 3 flow rate(s),liquid 3′ local components, volume ofliquid 3′ locally subjected to various fields in thetrough member 30 a′, neighboring (e.g., both upstream and downstream) electrode sets, local field concentrations, the use and/or position and/or composition of any membrane used in the trough member, etc., are all factors which influence processing conditions as well as composition and/or volume of constituents produced in the liquid 3′, nanocrystals and nanocrystal/suspensions or colloids made according to the various embodiments disclosed herein. Accordingly, a plethora of embodiments can be practiced according to the detailed disclosure presented herein. - The wires used to attach
electrode 1 to thetransformer 60 were, for Examples 1-3, 99.95% (3N5) gold wire, having a diameter of about 1 mm. Theplasma 4 was created with anelectrode 1 similar in shape to that shown inFIG. 3 e , and weighed about 9.2 grams. This electrode was 99.95% pure gold. Theother electrode 5 a measured about 1 mm thick gold wire (99.95%) and having about 9 mm submerged in the liquid 3′. - As shown in
FIGS. 10 b and 11 a , the output from thetrough member 30 a′ was the conditionedliquid 3′ and this conditioned liquid 3′ flowed directly into asecond trough member 30 b′. Thesecond trough member 30 b′, shown inFIGS. 10 b and 11 a had measurements as reported in Table 1. Thistrough member 30 b′ contained about 885 ml ofliquid 3″. Table 1 reports the electrode configuration, as shown inFIGS. 8 b and 11 a , which means seven sets ofelectrodes 5/5′ (shown inFIG. 8 b ) were positioned as shown inFIG. 11 a (i.e., perpendicular to the flow direction of the liquid 3″). Each of the electrode sets 5/5′ comprised 99.99% pure gold wire measuring about 1.0 mm in diameter, as reported in Table 1. The length of eachwire electrode 5 that was in contact with the liquid 3″ (reported as “WL” in Table 1) measured about 1″ (about 25.4 mm). Other orientations fit within the metes and bounds of this disclosure. - The AC power source (or transformer) 501AC, illustrated in
FIG. 13 , was used as the power supply for examples contained herein. Thistransformer 501 AC was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of about 2 kVA. With regard toFIGS. 10 a-10 d and 11 a-11 b , each separate electrode set 5/5′ (e.g.,Set 2, Set 3-Set 8 or Set 9) were electrically connected to the power supply 501AC as shown inFIG. 10 a . Specifically, power supply 501AC was electrically connected to each electrode set, according to the wiring diagram show inFIG. 10 a . Table 1 refers to each of the electrode sets by “Set #” (e.g., “Set 1” through “Set 8”). Each electrode of the 1/5 or 5/5 electrode sets was set to operate at a specific voltage. The voltages listed in Table 1 are the voltages used for each electrode set. The distance “c-c” (with reference toFIG. 6 ) from the centerline of each electrode set to the adjacent electrode set is also reported. Further, the distance “x” associated with eachelectrode 1 utilized is also reported. For theelectrode 5, no distance “x” is reported. Other relevant parameters are also reported in Table 1. All materials for theelectrodes 1/5 were obtained from Hi-Rel having an address of 23 Lewis Street, Fort Erie, Ontario, Canada, L2A 2P6. With reference toFIGS. 10 b, 10 c and 11 a , eachelectrode 5/5′ was first placed into contact with the liquid 3″ such that it just entered the female receiver tube o5. After a certain amount of process time, gold metal was removed from eachwire electrode 5 which caused theelectrode 5 to thin (i.e., become smaller in diameter) which changed, for example, current density and/or the rate at which gold nanoparticles were formed. Accordingly, theelectrodes 5 were moved toward the female receiver tubes o5 resulting in fresh andthicker electrodes 5 entering the liquid 3″ at a top surface portion thereof. In essence, an erosion profile or tapering effect was formed on theelectrodes 5 after some amount of processing time has passed (i.e., portions of the wire near the surface of the liquid 3″ were typically thicker than portions near the female receiver tubes o5), and such wire electrode profile or tapering can remain essentially constant throughout a production process, if desired, resulting in essentially identical product being produced at any point in time after an initial pre-equilibrium phase during a production run allowing, for example, the process to be cGMP under current FDA guidelines and/or be ISO 9000 compliant as well. - The
electrodes 5/5 were actuated or moved at a rate of about 1 inch per 8 hours. Samples were collected only from the equilibrium phase. The pre-equilibrium phase occurs because, for example, the concentration of nanocrystals produced in theliquid 3″ increases as a function of time until the concentration reaches equilibrium conditions (e.g., substantially constant nucleation and growth conditions within the apparatus), which equilibrium conditions remain substantially constant through the remainder of the processing due to the control processes disclosed herein. - The eight
electrode sets 1/5 and 5/5 were all connected to controldevices 20 through 20 g which automatically adjusted the height of, for example, eachelectrode 1/5 or 5/5 in each electrode set. Two female receiver tubes o5 a/o5 a′-o5 g/o5 g′ were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. Each female receiver tube o5 was made of polycarbonate and had an inside diameter of about ⅛ inch (about 3.2 mm) and was fixed in place by a solvent adhesive to the bottom portion of thetrough member 30 b′. Holes in the bottom of thetrough member 30 b′ permitted the outside diameter of each tube o5 to be fixed therein such that one end of the tube o5 was flush with the surface of the bottom portion of thetrough 30 b′. The bottom portion of the tube o5 is sealed. The inside diameters of the tubes o5 effectively prevented any significant quantities ofliquid 3″ from entering into the female receiver tube o5. However, some liquid may flow into the inside of one or more of the female receiver tubes o5. The length or vertical height of each female receiver tube o5 used in this Example was about 6 inches (about 15.24 cm) however, shorter or longer lengths fall within the metes and bounds of this disclosure. Further, while the female receiver tubes o5 are shown as being subsequently straight, such tubes could be curved in a J-shaped or U-shaped manner such that their openings away from thetrough member 30 b′ could be above the top surface of theliquid 3,” if desired. - The run described in this example utilize the following processing enhancer, Specifically, about 2.0 grams/gallon (i.e., about 0.528 g/liter) of sodium hydrogen carbonate (“soda”), having a chemical formula of NaHCO3, was added to and mixed with the
water 3. The soda was obtained from Alfa Aesar and the soda had a formula weight of 84.01 and a density of about 2.159 g/cm3. - In particular, a sine wave AC frequency at 60 Hz was utilized to make nanocrystal suspensions or colloids and/or ion solutions in accordance with the teachings herein. The AC power source 501AC utilized a Chroma 61604 programmable AC source. The applied voltage was about 220 volts. The applied current was between about 4.5 amps and about 5.5 amps.
- Table 1 summarizes key processing parameters used in conjunction with
FIGS. 9 and 10 c. Also, Table 1 discloses: 1) “Produced Au PPM” (e.g., gold nanocrystal concentrations); 2) “TEM Average Diameter” which is the mode, corresponding to the crystal diameter that occurs most frequently, determined by the TEM analysis; and 3) “Hydrodynamic radius” as measured by the Zetasizer ZS-90. These physical characterizations were performed as discussed elsewhere herein. - Specifically, TEM samples were prepared by utilizing a Formvar coated grid stabilized with carbon having a mesh size of 200. The grids were first pretreated by a plasma treatment under vacuum. The grids were placed on a microscope slide lined with a rectangular piece of filter paper and then placed into a Denton Vacuum apparatus with the necessary plasma generator accessory installed. The vacuum was maintained at 75 mTorr and the plasma was initiated and run for about 30 seconds. Upon completion, the system was vented and the grids removed. The grids were stable up to 7-10 days depending upon humidity conditions, but in all instances were used within 12 hours.
- Approximately 1 μL of each inventive nanocrystal suspension was placed onto each grid and was allowed to air dry at room temperature for 20-30 minutes, or until the droplet evaporated. Upon complete evaporation, the grids were placed onto a holder plate until TEM analysis was performed.
- A Philips/
FEI Tecnai 12 Transmission Electron Microscope was used to interrogate all prepared samples. The instrument was run at an accelerating voltage of 100 keV. After alignment of the beam, the samples were examined at various magnifications up to and including 630,000×. Images were collected via the attached Olympus Megaview III side-mounted camera that transmitted the images directly to a PC equipped with iTEM and Tecnai User Interface software which provided for both control over the camera and the TEM instrument, respectively. -
FIG. 11 c shows a representative TEM photomicrograph corresponding to dried solution NE10214 comprised of gold nanocrystals, dried from suspension, made according to this example.FIG. 11 d corresponds to the measured TEM size distribution used to calculate the TEM average diameter and referenced in Table 1. - The pH measurements were made by using an Accumet® AR20 pH/conductivity meter wherein the pH probe was placed into a 50 mL vial containing the samples of interest and allowed to stabilize. Three separate pH measurements were then taken and averaged per sample. NE10214 had a pH of about 8.94.
- Energy absorption spectra were obtained for the samples by using UV-VIS spectroscopy. This information was acquired using a
Thermofisher Evolution 201 UV-VIS spectrometer equipped with a double beam Czerny-Turner monochromator system and dual silicon photodiodes. Instrumentation was provided to support measurement of low-concentration liquid samples using one of a number of fuzed-quartz sample holders or “cuvettes.” Data was acquired over the wavelength range between about 300-900 nm with the following parameters: bandwidth of 1 nm, data pitch of 0.5 nm. A xenon flash lamp was the primary energy source. The optical pathway of the spectrometer was arranged to allow the energy beam to pass through the center of each sample cuvette. Sample preparation was limited to filling and capping the cuvettes and then physically placing the samples into the cuvette holder, within the fully enclosed sample compartment of the spectrometer. Optical absorption of energy of each sample was determined. Data output was measured and displayed as Absorbance Units (per Beer-Lambert's Law) versus wavelength. -
FIG. 11 e shows UV-Vis spectral patterns for the suspension/colloid NE10214, for the wavelength range of about 350 nm-900 nm. - Specifically, dynamic light scattering (DLS) measurements were performed on Zetasizer Nano ZS-90 DLS instrument. In DLS, as the laser light hits small particles and/or organized water structures around the small particles (smaller than the wavelength), the light scatters in all directions, resulting in a time-dependent fluctuation in the scattering intensity. Intensity fluctuations are due to the Brownian motion of the scattering particles/water structure combination and contain information about the crystal size distribution.
- The instrument was allowed to warm up for at least 30 min prior to the experiments. The measurements were made using square glass cell with 1 cm pathlength, PCS8501. The following procedure was used:
-
- 1. First, 1 ml of DI water was added into the cell using 1 ml micropipette, then water was poured out of the cell to a waste beaker and the rest of the water was shaken off the cell measuring cavity. This step was repeated two more times to thoroughly rinse the cell.
- 2. 1 ml of the sample was added into the cell using 1 ml micropipette. After that all liquid was removed out of the cell with the same pipette using the same pipette tip and expelled into the waste beaker. 1 ml of the sample was added again using the same tip.
- 3. The cell with the sample was placed into a temperature controlled cell block of the Zetasizer instrument with engraved letter facing forward. A new experiment in Zetasizer software was opened. The measurement was started 1 min after the temperature equilibrated and the laser power attenuated to the proper value. The results were saved after all runs were over.
- 4. The cell was taken out of the instrument and the sample was removed out of the cell using the same pipette and the tip used if
step 2. - 5.
Steps 2 to 4 were repeated two more times for each sample. - 6. For a new sample, a new pipette tip for 1 ml pipette was taken to avoid contamination with previous sample and
steps 1 through 5 were repeated.
Data collection and processing was performed with Zetasizor software, version 6.20. The following parameters were used for all the experiments: Run Duration—2o; Experiments—10; Solvent—water, 0 mmol; Viscosity—0.8872 cP; Refractive Index—1.333; block temperature—+25° C. After data for each experiment were saved, the results were viewed on “Records View” page of the software. Particle size distribution (i.e., hydrodynamic radii) was analyzed in “Intensity PSD” graph. Dynamic light scattering techniques were utilized to obtain an indication of crystal sizes (e.g., hydrodynamic radii) produced according to this example. Hydrodynamic radius is reported in Table 1 as 19.43 nm.
- The AAS values were obtained from a Perkin Elmer AAnalyst 400 Spectrometer system. Atomic absorption spectroscopy is used to determine concentration of species, reported in “ppm” (parts per million).
-
-
- The technique of flame atomic absorption spectroscopy requires a liquid sample to be aspirated, aerosolized and mixed with combustible gases, such as acetylene and air. The mixture is ignited in a flame whose temperature ranges from about 2100 to about 2400 degrees C. During combustion, atoms of the element of interest in the sample are reduced to free, unexcited ground state atoms, which absorb light at characteristic wavelengths. The characteristic wavelengths are element specific and are accurate to 0.01-0.1 nm. To provide element specific wavelengths, a light beam from a hollow cathode lamp (HCL), whose cathode is made of the element being determined, is passed through the flame. A photodetector detects the amount of reduction of the light intensity due to absorption by the analyte. A monochromator is used in front of the photodetector to reduce background ambient light and to select the specific wavelength from the HCL required for detection. In addition, a deuterium arc lamp corrects for background absorbance caused by non-atomic species in the atom cloud.
-
-
- 10 mL of sample, 0.6 mL of 36% v/v hydrochloric acid and 0.15 mL of 50% v/v nitric acid are mixed together in a glass vial and incubated for about 10 minutes in 70 degree C. water bath. If gold concentration in the suspension is expected to be above 10 ppm a sample is diluted with DI water before addition of the acids to bring final gold concentration in the range of 1 to 10 ppm. For example, for a gold concentration around 100 ppm, 0.5 mL of sample is diluted with 9.5 mL of DI water before the addition of acids. Aliquoting is performed with adjustable micropipettes and the exact amount of sample, DI water and acids is measured by an Ohaus PA313 microbalance. The weights of components are used to correct measured concentration for dilution by DI water and acids.
- Each sample is prepared in triplicate and after incubation in water bath is allowed to cool down to room temperature before measurements are made.
-
-
- The following settings are used for Perkin Elmer AAnalyst 400 Spectrometer system:
- a) Burner head: 10 cm single-slot type, aligned in three axes according to the manufacture procedure to obtain maximum absorbance with a 2 ppm Cu standard.
- b) Nebulizer: plastic with a spacer in front of the impact bead.
- c) Gas flow: oxidant (air) flow rate about 12 L/min, fuel (acetylene) flow rate about 1.9 mL/min.
- d) Lamp/monochromator: Au hollow cathode lamp, 10 mA operating current, 1.8/1.35 mm slits, 242.8 nm wavelength, background correction (deuterium lamp) is on.
-
-
- a) Run the Au lamp and the flame for approximately 30 minutes to warm up the system.
- b) Calibrate the instrument with 1 ppm, 4 ppm and 10 ppm Au standards in a matrix of 3.7% v/v hydrochloric acid. Use 3.7% v/v hydrochloric acid as a blank.
- c) Verify calibration scale by measuring 4 ppm standard as a sample. The measured concentration should be between 3.88 ppm and 4.12 ppm. Repeat step b) if outside that range.
- d) Measure three replicas of a sample. If the standard deviation between replicas is higher than 5%, repeat measurement, otherwise proceed to the next sample.
- e) Perform verification step c) after measuring six samples or more often. If verification fails, perform steps b) and c) and remeasure all the samples measured after the last successful verification.
-
-
- Measured concentration value for each replica is corrected for dilution by water and acid to calculate actual sample concentration. The reported Au ppm value is the average of three corrected values for individual replica.
- Table 1 references the AAS concentration result as “Produced Au PPM”, with a corresponding value of 6.6 ppm
- This Example utilized a batch process according to the present invention.
FIG. 12 a shows the apparatus used to condition theliquid 3. Once conditioned, theliquid 3′ was processed in the apparatus shown inFIG. 12 c - The amount of NaHCO3 processing enhancer used was about 0.375 grams/gallon (i.e., about 0.10 g/L) to about 3.0 grams/gallon (i.e., about 0.79 g/L). The amount of KOH processing enhancer used was about 0.95 grams/gallon (i.e., about 0.25 g/L). The amount of KBr processing enhancer used was about 4.6 grams/gallon (i.e., about 1.22 g/L). The amount of Na3PO4 processing enhancer used was about 3.94 grams/gallon (i.e., about 1.04 g/L). The amount of KH2PO4 processing enhancer was about 3.24 grams/gallon (i.e., about 0.86 g/L). The amount of time that the
water 3 with processing enhancer was exposed to theplasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown inFIG. 12 c. - The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - A second and different transformer was electrically connected to the
electrodes 5 a/5 b shown inFIG. 12 c . This transformer was an by AC power source having a voltage range of 0-300V, a frequency range of 47-400 Hz and a maximum power rating of 1 kVA. The applied voltage ranged between about 58 volts and about 300 volts. The diameter of the platinum wire electrodes was either about 0.5 mm or 1 mm. - Another power supply was utilized for those processes with frequency between 1 and 5 Hz, inclusive. The
electrodes FIG. 12 e . The power supply for the amplifier is set forth inFIG. 12 f . The power amplifier was driven by an external function generator connected to the input pins in the amplifier. - The amount of platinum nanoparticles produced in the suspensions varied between about 10 ppm and about 25 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. The sizes of the nanoparticles made according to this Example are fully discussed in Tables 2 and 3 herein.
- Transmission electron microscopy (TEM) sample preparation was identical to the methods described earlier although interrogation was performed on a Philips EM 420 TEM equipped with a SIS Megaview III CCD digital camera. The TEM micrographs show that the particles have an average diameter of less than 10 nm.
-
FIG. 14 shows a representative TEM Photomicrograph of platinum nanocrystals, dried from suspension GRPt-621, made according to this example. -
TABLE 2 Potential, Container Liquid Diameter, Peak to Frequency Volume Volume pH, GZA WL 5a & 5b pH, GRPt Peak (V) (Hz) t (min) (mL) (mL) Processing Enhancer Liquid (min) (cm) (mm) ppm Final 601 76 1 60 600 400 2.0 g/gal NaHCO3 ** 8.6 30 2 0.5 13.3 9.1 602 100 1 94 600 400 2.0 g/gal NaHCO3 ** 8.6 30 2 0.5 16.8 9.3 603 69.6 1 182 600 450 2.0 g/gal NaHCO3 ** 8.6 30 2.9 0.5 24.5 9.2 605 128 1 11 600 400 2.0 g/gal NaHCO3 ** 8.6 30 4 0.5 11.6 8.7 6a 58.4 1 14 10 5 0.75 g/gal NaHCO3 8.6 30 2 0.5 18.7 606 128 1 32 600 400 0.75 g/gal NaHCO3 8.6 30 4 0.5 17.9 8.6 607 128 1 51 600 400 0.375 g/gal NaHCO3 8.6 30 4 0.5 16.3 8.2 611 130 1 51 600 400 0.375 g/gal NaHCO3 8.6 30 2 0.5 12.8 7.8 612 130 1 56 600 400 0.375 g/gal NaHCO3 8.6 30 2 0.5 15.8 8.1 613 130 1 40 600 400 0.375 g/gal NaHCO3 8.6 30 2 0.5 12.8 7.9 614a 128 5 24 600 400 3 g/gal NaHCO3 8.6 30 3.2 1 11.1 9.0 614b 128 1 24 600 400 3 g/gal NaHCO3 8.6 30 3.2 1 12.6 9.4 614c 128 0.5 29 600 400 3 g/gal NaHCO3 8.6 30 3.2 1 10.5 9.4 614di 128 3 24 600 400 3 g/gal NaHCO3 8.6 30 3.2 1 12.1 9.1 615a 130 1 (square) 23 600 400 3.24 g/gal KH2PO4 4.9 n/a 3.2 1 10.3 5.1 615b 130 1 26 600 400 3.24 g/gal KH2PO4 4.9 n/a 3.2 1 10.4 4.9 616 130 1 (square) 16 600 400 3 g/gal NaHCO3 8.6 n/a 3.2 1 16.8 9.5 619 104 1 25 600 400 3.94 g/gal Na3PO4 ** 11.4 n/a 3.2 1 12.7 11.5 620 130 2 20 150 100 0.95 g/gal KOH ** 11.7 n/a 3.2 1 16.7 11.6 621 104 2 24 150 100 4.6 g/gal KBr ** 6.3 n/a 3.2 1 23.7 9.4 622 90 2 41 150 100 1:1 4.6 g/gal KBr: 11.2 n/a 3.2 1 24.5 11.2 0.95 g/gal KOH ** -
TABLE 3 Container Liquid Diameter, Frequency Volume Volume Processing pH, GZA WL 5a & 5b pH, Lot Number Voltage (Hz) t (min) (mL) (mL) Enhancer Liquid (min) (cm) (mm) ppm Final CAC-002-1 100 1 35 1000 800 4 g/gal NaHCO3 8.5 30 1.9 1 22.9 n/m CAC-001-2 100 1 35 1000 800 4 g/gal NaHCO3 8.5 30 1.9 1 10.5 n/m CAC-003-2 170 1 35 1000 800 3 g/gal NaHCO3 8.5 30 1.9 1 9.3 n/m CAC-003-3 230 1 35 1000 800 2 g/gal NaHCO3 8.5 30 1.9 1 9.7 n/m CAC-003-6 300 1 35 1000 800 1 g/gal NaHCO3 8.5 30 1.9 1 7.9 n/m CAC-001-3 100 7 35 1000 800 4 g/gal NaHCO3 8.5 30 1.9 1 11.4 n/m CAC-002-4 100 15 35 1000 800 4 g/gal NaHCO3 8.5 30 1.9 1 10.4 n/m 071210-1 100 47 35 1000 800 4 g/gal NaHCO3 8.5 30 1.9 1 6.9 n/m 071210-2 100 60 35 1000 800 4 g/gal NaHCO3 8.5 30 1.9 1 7.2 n/m CAC-003-1 170 60 35 1000 800 3 g/gal NaHCO3 8.5 30 1.9 1 6.5 n/m CAC-003-4 230 60 35 1000 800 2 g/gal NaHCO3 8.5 30 1.9 1 9.2 n/m CAC-003-5 300 60 35 1000 800 1 g/gal NaHCO3 8.5 30 1.9 1 8.4 n/m 070110-3 100 100 35 1000 800 4 g/gal NaHCO3 8.5 30 1.9 1 6.6 n/m 071310-4 100 200 35 1000 800 4 g/gal NaHCO3 8.5 30 1.9 1 7.6 n/m - This Example utilized a batch process according to the present invention.
FIG. 12 a shows the apparatus used to condition theliquid 3. Once conditioned, theliquid 3′ was processed in the apparatus shown inFIG. 12 d. - The amount of KBr processing enhancer used was about 4.6 grams/gallon (i.e., about 1.2 grams/Liter) or about 1.4 g/gal (i.e., about 0.4 g/L). The amount of Na3PO4 processing enhancer used was about 1.9 grams/gallon (i.e., about 0.5 g/L). The amount of time that the
water 3 with each processing enhancer was exposed to theplasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown inFIG. 12 d. - The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - A power supply (shown in
FIG. 12 f ) was utilized to apply a sinusoidal voltage with a frequency of about 2.5 Hz to theelectrodes FIG. 12 e . The distance between the electrodes was fixed in all suspensions at approximately 7 mm. The amplifier was driven by an external function generator connected to the input pins in the amplifier. - The amount of platinum-based nanoparticles and/or platinum based ions produced in the suspensions was measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. Suspensions PRX37-01 and PRX37-02 show that for a given conductivity of
water 3, and a given voltage applied at a fixed distance toelectrodes - The average hydrodynamic radii of the formed particles in water were analyzed with the dynamic light scattering technique discussed elsewhere herein. The hydrodynamic radius is not reported (NR) for formulation PRX37-02 because the transmission amount reported in the DLS device was 100%, indicating a high presence of dissolved platinum species (e.g., ions).
- Transmission electron microscopy (TEM) sample preparation was identical to the methods described earlier although interrogation was performed on a Philips EM 420 TEM equipped with a SIS Megaview III CCD digital camera. PRX37-03 was the only formulation analyzed by TEM. The TEM micrographs show that the particles in suspension in formulation PRX37-03 had an average diameter of approximately 7 nm. The distribution of particle size is shown in
FIG. 15 b .FIG. 15 a shows a representative TEM Photomicrograph of platinum nanocrystals, dried from suspension PRX37-03, made according to this Example 3. Table 4 is included to show the relevant processing conditions used as well as certain resultant physical properties of the formulation PRX37. -
TABLE 4 PRX37 01 02 03 Potential, Peak 50 50 75 to Peak (V) Frequency 2.5 2.5 2.5 (Hz) t (min) 1250 1320 1370 Liquid Volume 3800 3800 3800 (mL) Processing 4.6 g/gal 1.9 g/gal 1.4 g/gal Enhancer KBr Na3PO4, KBr 1.4 g/gal KBr GZA (min) 30 30 30 pH, Liquid 3.8 11.3 3.8 Conductivity 1.6 1.6 0.7 (mS/cm) WL (cm) 3.8 3.8 3.8 Diameter, 5a 0.05 0.05 0.05 & 5b (cm) rhydro (nm) 15 NR 9 (global max.) rTEM (nm) NM NM 7 (global max.) ppm 40.3 22.5 22.1 pH, Final 4.3 11.2 4.0 - In general, this Example utilizes certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 d and 11 b. The AC power source (or transformer) 501AC, illustrated inFIG. 13 , was used as the power supply for the examples contained herein, while the function generator 501FG was sometimes used (as disclosed herein) to drive the AC power source 501AC. Thistransformer 501 AC was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of about 2 kVA. The precise electrical connections are discussed elsewhere herein.Control devices 20, as illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively. However, due to the short run times in each “Run ID,” there was no need to actuate thecontrol devices 20. Thus, theends 9′ of theelectrodes trough member 30 b′. - The amount of NaHCO3(Fisher Scientific, Cat #S631-3) processing enhancer used was about 2.5 grams/gallon (i.e., about 0.67 g/L) to about 3.5 grams/gallon (i.e., about 0.93 g/L). The amount of KHCO3 processing enhancer used was about 2.31 grams/gallon (i.e., about 0.61 g/L). The amount of NaOH processing enhancer used was about 0.70 grams/gallon (i.e., about 0.19 g/L). The amount of KOH processing enhancer used was about 0.72 grams/gallon (i.e., about 0.19 g/L). The amount of NaBr processing enhancer was about 2.18 grams/gallon (i.e., about 0.58 g/L). The amount of KBr processing enhancer was about 2.04 grams/gallon (i.e., about 0.54 g/L). The amount of Na2PO4 processing enhancer was about 1.08 grams/gallon (i.e., about 0.29 g/L). The amount of NaCl processing enhancer was about 1.27 grams/gallon (i.e., about 0.34 g/L). The amount of CaCl2 processing enhancer was about 1.16 grams/gallon (i.e., about 0.31 g/L).
- The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - The AC power source 501AC utilized a Chroma 61604 programmable unit. In particular, sine wave AC frequencies at 5 Hz and 80 Hz were utilized to make nanocrystal suspensions or colloids and/or ions, in accordance with the teachings herein. The applied voltage was about 175 volts. Additionally, the function generator 501FG provided sine waves at frequencies less than 15 Hz to the AC power source 501AC, which subsequently amplified the input signal to about 175 volts at different frequencies. The applied current varied between about 3.0 amps and about 6.5 amps.
- Transmission electron microscopy (TEM) sample preparation methods were identical to the methods described earlier herein, although the interrogations were performed on a
FEI Tecnai 12 TEM equipped with a SIS Megaview III CCD digital camera. The TEM micrographs show that the formed particles have an average diameter of less than 10 nm.FIG. 16 shows a representative TEM Photomicrograph of platinum nanocrystals, dried from suspension PB-13, made according to this Example 4. - The amount of platinum nanoparticles or ions produced in the formulations varied between about 1.0 ppm and about 15 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Tables 5-8 summarize key processing parameters used in conjunction with
FIGS. 9 a and 10 d . Also, Tables 5-8 disclose: 1) resultant “ppm” (e.g., platinum nanocrystal/ion concentrations.) - Note, while two different chlorine-based processing enhancers were used to make platinum species in water, a variety of issues exist when making gold-based nanocrystal suspensions which render them less than desirable for Au—Pt nanocrystal suspensions.
-
TABLE 5 Run ID: PB-09 PB-10/PB-13 PB-16 PB-17 PB-18 PB-19 Flow In (ml/min) 220 220 220 220 220 220 Rate: Out (ml/min) 200 200 200 200 200 200 Volts: Set # 1750 750 750 750 750 750 Set #'s 2-8 175 175 175 175 175 175 Set #'s 2-8 frequency, Hz 80 5 80 5 80 5 PE/Concentration (mg/ml) NaHCO3/0.67 NaHCO3/0.67 KHCO3/0.61 KHCO3/0.61 K2CO3/0.33 K2CO3/0.33 Wire Diameter (mm) 1.0 1.0 1.0 1.0 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b 8b 8b 8b 8b Produced Pt PPM 8.1 11.8 2.3 5.9 2.4 7.0 Output Temp ° C. at 32 70 70 65 63 66 64 Dimensions Plasma 4 Figs. 9 9 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm)36/914 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 S (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 Electrode Curr. (A) 0.72 0.67 0.67 0.61 0.67 0.60 Total Curr. Draw (A) 5.00 n/m 4.64 4.78 4.70 4.79 “c-c” (mm) 76 76 76 76 76 76 Set electrode # 1a 1a 1a 1a 1a 1a 1 “x” (in/mm) 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 electrode # 5a 5a 5a 5a n/a 5a “c-c” (mm) 102 102 102 102 102 102 Set electrode # 5b 5b 5b 5b 5b 5b 2 “x” (in/mm) n/a n/a n/a n/a n/a n/a electrode # 5b′ 5b′ 5b′ 5b′ 5b′ 5b′ “c-c” (mm) 76 76 76 76 76 76 Set electrode # 5c 5c 5c 5c 5c 5c 3 electrode # 5c′ 5c′ 5c′ 5c′ 5c′ 5c′ “c-c” (mm) 76 76 76 76 76 76 Set electrode # 5d 5d 5d 5d 5d 5d 4 electrode # 5d′ 5d′ 5d′ 5d′ 5d′ 5d′ “c-c” (mm) 127 127 127 127 127 127 Set electrode # 5e 5e 5e 5e 5e 5e 5 electrode # 5e′ 5e′ 5e′ 5e′ 5e′ 5e′ “c-c” (mm) 127 127 127 127 127 127 Set electrode # 5f 5f 5f 5f 5f 5f 6 electrode # 5f′ 5f′ 5f′ 5f′ 5f′ 5f′ “c-c” (mm) 152 152 152 152 152 152 Set electrode # 5g 5g 5g 5g 5g 5g 7 electrode # 5g′ 5g′ 5g′ 5g′ 5g′ 5g′ “c-c” (mm) 178 178 178 178 178 178 Set electrode # 5h 5h 5h 5h 5h 5h 8 electrode # 5h′ 5h′ 5h′ 5h′ 5h′ 5h′ “c-c” (mm) 76 76 76 76 76 76 -
TABLE 6 Run ID: PB-20 PB-21 PB-23 PB-24 PB-25 PB-26 Flow In (ml/min) 220 220 220 220 220 220 Rate: Out (ml/min) 200 200 200 200 200 200 Volts: Set # 1750 750 750 750 750 750 Set #'s 2-8 175 175 175 175 175 175 Set #'s 2-8 frequency, Hz 80 5 80 5 80 5 PE/Concentration(mg/ml) Na2CO3/0.30 Na2CO3/0.30 NaOH/0.19 NaOH/0.19 KOH/0.19 KOH/0.19 Wire Diameter (mm) 1.0 1.0 1.0 1.0 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b 8b 8b 8b 8b Produced Pt PPM 2.4 7.0 1.1 3.6 1.4 3.9 Output Temp ° C. at 32 68 66 60 60 63 60 Dimensions Plasma 4 Figs. 9 9 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm)36/914 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 S (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 Electrode Curr. (A) 0.73 0.63 0.55 0.51 0.53 0.51 Total Curr. Draw (A) 5.09 4.95 3.83 3.67 4.11 3.63 “c-c” (mm) 76 76 76 76 76 76 Set electrode # 1a 1a 1a 1a 1a 1a 1 “x” (in/mm) 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 electrode # 5a 5a 5a 5a n/a 5a “c-c” (mm) 102 102 102 102 102 102 Set electrode # 5b 5b 5b 5b 5b 5b 2 “x” (in/mm) n/a n/a n/a n/a n/a n/a electrode # 5b′ 5b′ 5b′ 5b′ 5b′ 5b′ “c-c” (mm) 76 76 76 76 76 76 Set electrode # 5c 5c 5c 5c 5c 5c 3 electrode # 5c′ 5c′ 5c′ 5c′ 5c′ 5c′ “c-c” (mm) 76 76 76 76 76 76 Set electrode # 5d 5d 5d 5d 5d 5d 4 electrode # 5d′ 5d′ 5d′ 5d′ 5d′ 5d′ “c-c” (mm) 127 127 127 127 127 127 Set electrode # 5e 5e 5e 5e 5e 5e 5 electrode # 5e′ 5e′ 5e′ 5e′ 5e′ 5e′ “c-c” (mm) 127 127 127 127 127 127 Set electrode # 5f 5f 5f 5f 5f 5f 6 electrode # 5f′ 5f′ 5f′ 5f′ 5f′ 5f′ “c-c” (mm) 152 152 152 152 152 152 Set electrode # 5g 5g 5g 5g 5g 5g 7 electrode # 5g′ 5g′ 5g′ 5g′ 5g′ 5g′ “c-c” (mm) 178 178 178 178 178 178 Set electrode # 5h 5h 5h 5h 5h 5h 8 electrode # 5h′ 5h′ 5h′ 5h′ 5h′ 5h′ “c-c” (mm) 76 76 76 76 76 76 -
TABLE 7 Run ID: PB-27 PB-28 PB-32 PB-33 PB-34 PB-35 Flow In (ml/min) 220 220 220 220 220 220 Rate: Out (ml/min) 200 200 200 200 200 200 Volts: Set # 1750 750 750 750 750 750 Set #'s 2-8 175 175 175 175 175 175 Set #'s 2-8 frequency, Hz 80 5 80 5 80 5 PE/Concentration(mg/ml) NaBr/0.58 NaBr/0.58 KBr/0.54 KBr/0.54 Na2PO4/0.29 KOH/0.29 Wire Diameter (mm) 1.0 1.0 1.0 1.0 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b 8b 8b 8b 8b Produced Pt PPM 2.5 9.9 2.2 7.1 1.6 4.1 Output Temp ° C. at 32 68 70.5 61.5 64 61 61 Dimensions Plasma 4 Figs. 9 9 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm)36/914 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 S (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 Electrode Curr. (A) 0.70 0.73 0.70 0.68 0.47 0.55 Total Curr. Draw (A) 4.88 5.31 3.95 4.14 4.03 4.43 “c-c” (mm) 76 76 76 76 76 76 Set electrode # 1a 1a 1a 1a 1a 1a 1 “x” (in/mm) 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 electrode # 5a 5a 5a 5a n/a 5a “c-c” (mm) 102 102 102 102 102 102 Set electrode # 5b 5b 5b 5b 5b 5b 2 “x” (in/mm) n/a n/a n/a n/a n/a n/a electrode # 5b′ 5b′ 5b′ 5b′ 5b′ 5b′ “c-c” (mm) 76 76 76 76 76 76 Set electrode # 5c 5c 5c 5c 5c 5c 3 electrode # 5c′ 5c′ 5c′ 5c′ 5c′ 5c′ “c-c” (mm) 76 76 76 76 76 76 Set electrode # 5d 5d 5d 5d 5d 5d 4 electrode # 5d′ 5d′ 5d′ 5d′ 5d′ 5d′ “c-c” (mm) 127 127 127 127 127 127 Set electrode # 5e 5e 5e 5e 5e 5e 5 electrode # 5e′ 5e′ 5e′ 5e′ 5e′ 5e′ “c-c” (mm) 127 127 127 127 127 127 Set electrode # 5f 5f 5f 5f 5f 5f 6 electrode # 5f′ 5f′ 5f′ 5f′ 5f′ 5f′ “c-c” (mm) 152 152 152 152 152 152 Set electrode # 5g 5g 5g 5g 5g 5g 7 electrode # 5g′ 5g′ 5g′ 5g′ 5g′ 5g′ “c-c” (mm) 178 178 178 178 178 178 Set electrode # 5h 5h 5h 5h 5h 5h 8 electrode # 5h′ 5h′ 5h′ 5h′ 5h′ 5h′ “c-c” (mm) 76 76 76 76 76 76 -
TABLE 8 Run ID: PB-40 PB-41 PB-42 PB-43 Flow In (ml/min) 220 220 220 220 Rate: Out (ml/min) 200 200 200 200 Volts: Set # 1750 750 750 750 Set #'s 2-8 175 175 175 175 Set #'s 2-8 frequency, Hz 80 5 80 5 PE/Concentration (mg/ml) NaCl/0.34 NaCl/0.34 CaCl2/0.31 CaCl2/0.31 Wire Diameter (mm) 1.0 1.0 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b 8b 8b Produced Pt PPM 1.5 10.2 2.0 2.0 Output Temp ° C. at 32 69 70.5 72 72 Dimensions Plasma 4 Figs. 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm)36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 S (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 Electrode Curr. (A) 0.72 0.72 0.77 0.73 Total Curr. Draw (A) 5.00 6.08 5.36 5.77 “c-c” (mm) 76 76 76 76 Set 1 electrode # 1a 1a 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 electrode # 5a 5a 5a 5a “c-c” (mm) 102 102 102 102 Set 2 electrode # 5b 5b 5b 5b “x” (in/mm) n/a n/a n/a n/a electrode # 5b′ 5b′ 5b′ 5b′ “c-c” (mm) 76 76 76 76 Set 3 electrode # 5c 5c 5c 5c electrode # 5c′ 5c′ 5c′ 5c′ “c-c” (mm) 76 76 76 76 Set 4 electrode # 5d 5d 5d 5d electrode # 5d′ 5d′ 5d′ 5d′ “c-c” (mm) 127 127 127 127 Set 5 electrode # 5e 5e 5e 5e electrode # 5e′ 5e′ 5e′ 5e′ “c-c” (mm) 127 127 127 127 Set 6 electrode # 5f 5f 5f 5f electrode # 5f′ 5f′ 5f′ 5f′ “c-c” (mm) 152 152 152 152 Set 7 electrode # 5g 5g 5g 5g electrode # 5g′ 5g′ 5g′ 5g′ “c-c” (mm) 178 178 178 178 Set 8 electrode # 5h 5h 5h 5h electrode # 5h′ 5h′ 5h′ 5h′ “c-c” (mm) 76 76 76 76 - In general, this Example utilizes certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 d and 11 b. The AC power source (or transformer) 501AC, illustrated inFIG. 13 , was used as the power supply for the examples contained herein, while the function generator 501FG was sometimes used (as disclosed herein) to drive the AC power source 501AC. Thistransformer 501 AC was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of about 2 kVA. The precise electrical connections are discussed elsewhere herein.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively. However, due to the short run times in each “Run ID,” there was no need to actuate thecontrol devices 20. Thus, theends 9′ of theelectrodes trough member 30 b′. Each run in this example utilized about 2.5 g/gallon of NaHCO3 as a processing enhancer and a liquid flow rate of about 220 ml/min. - Moreover, to show the effect of different frequencies on the process and/or products formulated, varying sine wave frequencies were utilized. In particular, sine wave AC frequencies as low as about 1 Hz and as high as about 200 Hz were utilized to make nanocrystal suspensions or colloids and/or ions, in accordance with the teachings herein. The AC power source 501AC utilized a Chroma 61604 programmable AC source. The applied voltage was about 175 volts with a corresponding sine wave at six different frequencies of about 15, 40, 60, 80, 100 and 200 Hz. Additionally, the function generator 501FG provided sine waves at frequencies less than 15 Hz to the power supply 501AC which subsequently amplified the input signal to about 175V at different frequencies, namely 1 Hz and 5 Hz. The applied current varied between about 4.5 amps and 6.0 amps.
- The amount of platinum nanoparticles and/or ions produced in the formulations varied between about 7.0 ppm and about 15 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Tables 9-10 summarize key processing parameters used in conjunction with
FIGS. 9 and 10 d. Also, Tables 9-10 disclose: 1) resultant “ppm” (i.e., platinum concentrations.) - Energy absorption spectra were obtained for the samples by using UV-VIS spectroscopy methods as outlined elsewhere herein.
FIG. 17 contains the UV-Vis data collected for the samples above, specifically displaying the 265 nm-750 nm range. -
TABLE 9 Run ID: PB-01 PB-02 PB-03 PB-04 PB-05 PB-06 Flow In (ml/min) 220 220 220 220 220 220 Rate: Out (ml/min) 184 200 200 200 200 200 Volts: Set # 1750 750 750 750 750 750 Set #'s 2-8 175 175 175 175 175 175 Set #'s 2-8 frequency, Hz 60 40 15 1 5 80 PE: NaHCO3 (mg/ml) 0.67 0.67 0.67 0.67 0.67 0.67 Wire Diameter (mm) 1.0 1.0 1.0 1.0 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b 8b 8b 8b 8b Produced Pt PPM 9.7 8.6 8.7 12.1 14.6 7.7 Output Temp ° C. at 32 72 72 72 71 72 71 Dimensions Plasma 4 Figs. 9 9 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 L T (in/mm)36/914 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 1/25 S (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 Electrode Curr. (A) 0.77 0.77 0.76 0.32 0.71 0.75 Total Curr. Draw (A) 5.43 5.40 5.33 n/m n/m n/m “c-c” (mm) 76 76 76 76 76 76 Set electrode # 1a 1a 1a 1a 1a 1a 1 “x” (in/mm) 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 electrode # 5a 5a 5a 5a n/a 5a “c-c” (mm) 102 102 102 102 102 102 Set electrode # 5b 5b 5b 5b 5b 5b 2 “x” (in/mm) n/a n/a n/a n/a n/a n/a electrode # 5b′ 5b′ 5b′ 5b′ 5b′ 5b′ “c-c” (mm) 76 76 76 76 76 76 Set electrode # 5c 5c 5c 5c 5c 5c 3 electrode # 5c′ 5c′ 5c′ 5c′ 5c′ 5c′ “c-c” (mm) 76 76 76 76 76 76 Set electrode # 5d 5d 5d 5d 5d 5d 4 electrode # 5d′ 5d′ 5d′ 5d′ 5d′ 5d′ “c-c” (mm) 127 127 127 127 127 127 Set electrode # 5e 5e 5e 5e 5e 5e 5 electrode # 5e′ 5e′ 5e′ 5e′ 5e′ 5e′ “c-c” (mm) 127 127 127 127 127 127 Set electrode # 5f 5f 5f 5f 5f 5f 6 electrode # 5f′ 5f′ 5f′ 5f′ 5f′ 5f′ “c-c” (mm) 152 152 152 152 152 152 Set electrode # 5g 5g 5g 5g 5g 5g 7 electrode # 5g′ 5g′ 5g′ 5g′ 5g′ 5g′ “c-c” (mm) 178 178 178 178 178 178 Set electrode # 5h 5h 5h 5h 5h 5h 8 electrode # 5h′ 5h′ 5h′ 5h′ 5h′ 5h′ “c-c” (mm) 76 76 76 76 76 76 -
TABLE 10 Run ID: PB-07 PB-08 Flow In (ml/min) 220 220 Rate: Out (ml/min) 200 200 Volts: Set # 1750 750 Set #'s 2-8 175 175 Set #'s 2-8 100 200 frequency, Hz PE: NaHCO3 (mg/ml) 0.67 0.67 Wire Diameter (mm) 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b Produced Pt PPM 9.7 8.6 Output Temp ° C. at 32 71 71 Dimensions Plasma 4 Figs. 9 9 Process Figures 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 L T (in/mm)36/914 36/914 d (in/mm) 1/25 1/25 S (in/mm) 1.5/38 1.5/38 Electrode Curr. (A) 0.76 0.77 Total Curr. Draw (A) 5.24 5.33 “c-c” (mm) 76 76 Set 1electrode # 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 electrode # 5a 5a “c-c” (mm) 102 102 Set 2electrode # 5b 5b “x” (in/mm) n/a n/a electrode # 5b′ 5b′ “c-c” (mm) 76 76 Set 3electrode # 5c 5c electrode # 5c′ 5c′ “c-c” (mm) 76 76 Set 4electrode # 5d 5d electrode # 5d′ 5d′ “c-c” (mm) 127 127 Set 5electrode # 5e 5e electrode # 5e′ 5e′ “c-c” (mm) 127 127 Set 6electrode # 5f 5f electrode # 5f′ 5f′ “c-c” (mm) 152 152 Set 7electrode # 5g 5g electrode # 5g′ 5g′ “c-c” (mm) 178 178 Set 8electrode # 5h 5h electrode # 5h′ 5h′ “c-c” (mm) 76 76 - This Example utilizes a batch process according to the present invention.
FIG. 12 a shows the apparatus used to condition theliquid 3. Once conditioned, theliquid 3′ was processed in the apparatus shown inFIG. 12 c or 12 d, for platinum ions/particles and bi-metallic nanocrystals, respectively. The overall process of creating a bi-metallic nanocrystal suspension is described below and is summarized in Table 11. - Initially, platinum ions and/or particles were created in water by the following process. Approximately 4.0 grams/gallon (i.e., about 1.06 mg/mL) of processing enhancer baking soda (i.e., NaHCO3) was added to about 1 gallon of de-ionized water. The amount of time that the
water 3 with processing enhancer was exposed to theplasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown inFIG. 12 c. - The applied voltage for each
plasma 4 created atelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. Note that in Table 11 (and elsewhere herein) the reference to “GZA” is synonomous with creation ofplasma 4. - A second and different transformer was electrically connected to the
electrodes 5 a/5 b shown inFIG. 12 c . This transformer was a hy AC power source having a voltage range of about 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA. The applied voltage was about 100 volts with a frequency of about 60 hertz for approximately a 2-hour operating time. The diameter of the platinum wire electrodes was 1 mm. The length of the platinum wires was about 51 mm. - Subsequently, the platinum species and water formulation (raw material) prepared above was mixed with an equal amount of conditioned water, which conditioned
water 3′ was achieved with aplatinum electrode 1 creating aplasma 4 for about 30 minutes, and processing enhancer NaHCO3 0.5 g/gallon (0.132 mg/mL) NaHCO3) at a ratio of 1:1 to a total volume of about 800 mL. The liquid 3′ was then processed via the apparatus inFIG. 12 d with gold electrodes (99.99%, about 0.5 mm diameter and a length of about 6.25 in (15.88 cm) for about 40 minutes, with a hy AC power source having an applied voltage of about 160 volts and about 47 hertz. The hydrodynamic radius of the bi-metallic nanocrystals made was about 14.7 nm as measured by ViscoTek. The suspension contained about 16.1 ppm of Au and about 2.1 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. -
FIG. 18 shows a representative TEM Photomicrograph of the bi-metallic nanocrystal suspension dried from formulation 110910-4, which was made by techniques equivalent to those discussed elsewhere herein. - Energy absorption spectra was obtained for this sample (111710-a) using Uv-Vis spectroscopy methods as outlined elsewhere herein.
FIG. 12 g contains the UV-Vis data collected for this sample (111710-a), specifically displaying the 350-900 nm range. -
TABLE 11 Component 1Pretreatment-GZA Run Volume NaHCO3 time ID (mL) (grams) (hrs) 110910-2 3785 4 0.5 Pt ion treatment (Pt wires, 99.99%) Length Wire Volume Voltage Frequency Time of Wire Diameter (mL) (V) (Hz) (hrs) (in/cm) (mm) 3785 100 60 2 2.01/5.1 1 Component 2Pretreatment-Pt GZA Run Volume NaHCO3 time ID (mL) (grams) (hrs) N/A 3785 0.5 0.5 Composite Mix Mixture of Component 1 & 2Comp. Comp. Run 1 Vol. 2 Vol. Volume ID (mL) (mL) (mL) 111710-9 400 400 800 Gold Nanoparticle Treatment (Au wires, 99.99%) Length Wire Voltage Frequency Time Current of Wire Diameter (V) (Hz) (hrs) (A) (in/cm) (mm) 160 47 0.67 1.28 6.25/15.88 0.5 - Specifically, dynamic light scattering (DLS) measurements were performed on Viscotek 802 DLS instrument. In DLS, as the laser light hits small particles and/or organized water structures around the small particles (smaller than the wavelength), the light scatters in all directions, resulting in a time-dependent fluctuation in the scattering intensity. Intensity fluctuations are due to the Brownian motion of the scattering particles/water structure combination and contain information about the crystal size distribution.
- The instrument was allowed to warm up for at least 30 min prior to the experiments. The measurements were made using 12 μl quartz cell. The following procedure was used:
-
- 7. First, 1 ml of DI water was added into the cell using 1 ml micropipette, then water was poured out of the cell to a waste beaker and the rest of the water was shaken off the cell measuring cavity. This step was repeated two more times to thoroughly rinse the cell.
- 8. 100 μl of the sample was added into the cell using 200 μl micropipette. After that all liquid was removed out of the cell with the same pipette using the same pipette tip and expelled into the waste beaker. 100 μl of the sample was added again using the same tip.
- 9. The cell with the sample was placed into a temperature controlled cell block of the Viscotek instrument with frosted side of the cell facing left. A new experiment in Viscotek OmniSIZE software was opened. The measurement was started 1 min after the temperature equilibrated and the laser power attenuated to the proper value. The results were saved after all runs were over.
- 10. The cell was taken out of the instrument and the sample was removed out of the cell using the same pipette and the tip used if
step 2. - 11.
Steps 2 to 4 were repeated two more times for each sample. - 12. For a new sample, a new pipette tip for 200 μl pipette was taken to avoid contamination with previous sample and
steps 1 through 5 were repeated.
- Data collection and processing was performed with OmniSIZE software,
version - It should be noted that the dynamic light scattering particle size information is different from the TEM measured histograms because dynamic light scattering uses algorithms that assume the nanocrystals are all spheres (which they are not) as well as measures the hydrodynamic radius (e.g., the nanocrystal's influence on the water is also detected and reported in addition to the actual physical radii of the particles). Accordingly, it is not surprising that there is a difference in the reported particle sizes between those reported in the TEM histogram data and those reported in the dynamic light scattering data, just as in the other Examples included herein.
- This Example utilizes a batch process according to the present invention.
FIG. 12 a shows the apparatus used to condition theliquid 3. Once conditioned, theliquid 3′ was processed in the apparatus shown inFIG. 12 c or 12 d, for platinum ions/particles and bi-metallic nanocrystals, respectively. The overall process of creating a bi-metallic nanocrystal suspension is described below and is summarized in Table 12. - Initially, platinum ions and/or particles were created in water by the following process. Approximately 4.0 grams/gallon (i.e., about 1.06 mg/mL) of processing enhancer baking soda (i.e., NaHCO3) was added to about 1 gallon of de-ionized water. The amount of time that the
water 3 with processing enhancer was exposed to theplasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown inFIG. 12 c . Note that in Table 12 (and elsewhere herein) the reference to “GZA” is synonomous with creation ofplasma 4. - The applied voltage for each
plasma 4 created atelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - A second and different transformer was electrically connected to the
electrodes 5 a/5 b shown inFIG. 12 c . This transformer was a hy AC power source having a voltage range of 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA. The applied voltage was about 100 volts with a frequency of about 60 hertz for approximately a 2-hour operating time. The diameter of the platinum wire electrodes was about 1 mm. - Subsequently, the platinum species and water formulation (raw material) prepared above was mixed with about 6.29 mM NaHCO3 at a ratio of about 3:1 to create a total volume of about 3785 mL. This liquid 3′ was then processed via the apparatus shown in
FIG. 12 d with gold electrodes (99.99%, 0.5 mm) for about 90 minutes, with a hy AC power source having an applied voltage of about 200 volts and about 60 hertz. The hydrodynamic radius of the bi-metallic nanocrystals made was about 15.4 nm as measured by ViscoTek. The suspension contained about 5.6 ppm of Au and about 1.6 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. -
FIG. 19 shows a representative TEM Photomicrograph of the bi-metallic nanocrystal suspension dried from formulation 101910-6, which was obtained by techniques equivalent to those disclosed elsewhere herein. -
TABLE 12 Component 1Pretreatment-GZA Run Volume NaHCO3 time ID (mL) (grams) (hrs) 102910 3785 4 0.5 Pt ion treatment (Pt wires, 99.99%) Length Wire Volume Voltage Frequency Time of Wire Diameter (mL) (V) (Hz) (hrs) (in/cm) (mm) 3785 100 60 2 2.01/5.1 1 Component 22 g NaHCO3 (No GZA) Run Volume NaHCO3 time ID (mL) (grams) (hrs) N/A 3785 2.0 N/A Composite Mix Mixture of Component 1 & 2Comp. Comp. Run 1 Vol. 2 Vol. Volume ID (mL) (mL) (mL) 110810 946 2839 3785 Gold Nanoparticle Treatment (Au wires, 99.99%) Length Wire Voltage Frequency Time Current of Wire Diameter (V) (Hz) (hrs) (A) (in/cm) (mm) 200 60 1.5 1.07 6.25/15.88 0.5 - This Example utilizes a batch process according to the present invention.
FIG. 12 a shows the apparatus used to condition theliquid 3. Once conditioned, theliquid 3′ was processed in the apparatus shown inFIG. 12 c or 12 d, for platinum ions/particles and bi-metallic nanocrystals, respectively. The overall process of creating a bi-metallic nanocrystal suspension is described below and is summarized in Table 13. - Initially, platinum ions and/or particles were created in water by the following process. Approximately 0.580 grams/gallon (i.e., about 0.153 mg/mL) of processing enhancer potassium hydroxide (i.e., KOH) was added to about 1 gallon of de-ionized water. The amount of time that the
water 3 with processing enhancer was exposed to theplasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown inFIG. 12 c. - The applied voltage for each
plasma 4 created atelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. Note that in Table 13 (and elsewhere herein) the reference to “GZA” is synonomous with creation ofplasma 4. - A second and different transformer was electrically connected to the
electrodes 5 a/5 b shown inFIG. 12 c . This transformer was a hy AC power source having a voltage range of about 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA. The applied voltage was about 260 volts with a frequency of about 60 hertz for approximately a 2-hour operating time. The diameter of the platinum wire electrodes was about 1 mm. The length of the platinum wires was about 51 mm (2.01 inch/5.1 cm). - Subsequently, the platinum species and water formulation (raw material) prepared above was further processed as described below. The liquid 3′ was then processed via the apparatus in
FIG. 12 d with gold electrodes (99.99%, about 0.5 mm diameter and about 6.25 inches (15.88 cm) total length for about 2 hours, with a hy AC power source having an applied voltage of about 180 volts and about 47 hertz. The hydrodynamic radius of the gold/platinum material made was about 12.5 nm as measured by ViscoTek. The suspension contained about 8.0 ppm of Au and about 1.8 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. -
FIG. 20 shows a representative TEM Photomicrograph of the bi-metallic nanocrystal suspension dried from formulation ID #122310A, made according to this Example 8. -
TABLE 13 Component 1Pretreatment-GZA Run Volume KOH time ID (mL) (grams) (hrs) 122210-2 3785 0.580 0.5 Pt ion treatment (Pt wires, 99.99%) Length Wire Volume Voltage Frequency Time of Wire Diameter (mL) (V) (Hz) (hrs) (in/cm) (mm) 3785 260 60 2 2.01/5.1 1 Component 2 N/A Run Volume NaHCO3 time ID (mL) (grams) (hrs) N/A N/A N/A N/A Composite Mix Mixture of Component 1 & 2Comp. Comp. Run 1 Vol. 2 Vol. Volume ID (mL) (mL) (mL) 122310A 3785 0 3785 Gold Nanoparticle Treatment (Au wires, 99.99%) Length Wire Voltage Frequency Time Current of Wire Diameter (V) (Hz) (hrs) (A) (in/cm) (mm) 180 47 2.0 0.717 6.25/15.88 0.5 - This Example utilizes a batch process according to the present invention.
FIG. 12 a shows the apparatus used to condition theliquid 3. Once conditioned, theliquid 3′ was processed in the apparatus shown inFIG. 12 c or 12 d, for platinum ions/nanocrystal and for gold nanocrystals, respectively. The overall process of creating the individual nanocrystal suspensions and thus mixing them together to form a bi-metallic nanoparticle suspension is described below and is summarized in Table 14. - Initially, platinum ions and/or particles were created in water by the following process. Approximately 4.0 grams/gallon (i.e., about 1.06 mg/mL) of processing enhancer baking soda (i.e., NaHCO3) was added to about 1 gallon of de-ionized water. The amount of time that the
water 3 with processing enhancer was exposed to theplasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown inFIG. 12 c. - The applied voltage for each
plasma 4 created atelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - A second and different transformer was electrically connected to the
electrodes 5 a/5 b shown inFIG. 12 c . This transformer was a hy AC power source having a voltage range of about 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA. The applied voltage was about 130 volts with a frequency of about 60 hertz for approximately a 30-minute operating time. The diameter of the platinum wire electrodes was about 1 mm. The length of the platinum wires was about 51 mm. The platinum species and water material was set aside. - A separate suspension of gold nanocrystals was prepared as follows. Approximately 1.0 gram/gallon (i.e., about 0.264 mg/mL) of processing enhancer baking soda (i.e., NaHCO3) was added to about 1 gallon of de-ionized water. The amount of time that the
water 3 with processing enhancer was exposed to theplasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown inFIG. 12 c. - The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - A second and different transformer was electrically connected to the
electrodes 5 a/5 b shown inFIG. 12 d . This transformer was a hy AC power source having a voltage range of about 0-300V, a frequency range of about 47-400 Hz and a maximum power rating of about 1 kVA. The applied voltage was about 300 volts with a frequency of about 60 hertz for approximately a 30-minute operating time. The diameter of the gold wire electrodes was about 0.5 mm. The length of the gold wire was about 159 mm. - Subsequently, the separately prepared Pt and Au water-based materials Pt formulation and Au formulation prepared above were mixed together in the presence of a hydrogen peroxide catalyst (H2O2, Alfa Aesar Cat #L14000) and then studied. Specifically, about 300 mL of Pt formulation 062810 and about 700 mL of Au formulation 061610 were combined and approximately 250 μL of H2O2 0.8 v/v % was added. The measured hydrodynamic radius of the combined formulations was about 35 nm as measured by ViscoTek. The resulting suspension contained about 8.0 ppm of Au and about 1.8 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- A comparison of this suspension to a previously discussed bi-metallic nanoparticle suspension was then performed. Specifically, high resolution analysis and energy dispersive x-ray analysis indicated that the resultant colloids or suspensions had little to no platinum physically present between the formed gold nanocrystals, as shown in representative
FIGS. 23 a-23 b and in representative EDSFIGS. 24 a -24 b. - In contrast, sample 111710-9, made substantially identically to sample 112210-1 as described in Example 6, had identifiable platinum present on the formed bi-metallic nanocrystals. The measured hydrodynamic radius of the bi-metallic nanocrystals was about 14.7 nm as measured by ViscoTek. The suspension contained about 16.1 ppm of Au and about 2.1 ppm of Pt as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. Representative
FIGS. 21 a-21 b illustrate the structures formed when prepared as described above. It is evident through energy dispersive analysis that platinum is present at detectable concentrations, as indicated by representativeFIGS. 22 a -22 b. - TEM samples were prepared by utilizing a lacey Formvar/carbon-coated copper grid having a mesh size of 200. Approximately 1-3 μL of each inventive nanocrystal suspension, colloid and/or solution was placed onto each grid and was allowed to air dry at room temperature for about 20-30 minutes, or until the droplet evaporated. Upon complete evaporation, the grids were placed onto a holder plate until TEM analysis was performed.
- A Philips CM300 FEG High Resolution Transmission Electron Microscope, equipped with an Oxford thin window light element detector and
Emispec ES vision 4 processor, was used to interrogate all prepared samples. The instrument was run at an accelerating voltage of about 297 kV. After alignment of the electron beam, the prepared samples were examined at various magnifications up to and including 800,000×. Images were collected via the integrated CCD camera mounted at the back of the Gatan Image Filter (GIF) which is linked directly to a PC equipped with Digital Micrograph Software and Emispec ES Vision 4.0 software. Images were collected at a beam spot size of 2 corresponding to a beam width setting selected on the instrument and energy dispersive x-ray spectra were collected at a spot size of between 3-5, which allowed for the maximum amount of electrons to be collected. To increase the signal to noise ratio further, the Philips double-tilt holder was rotated 10 degrees towards the detector. Finally, the beam was condensed down to the area of interest and then the detector valve was opened and subsequent collection began. -
TABLE 14 Component 1-Pt solution Pretreatment-Au GZA Run Volume NaHCO3 time ID (mL) (grams) (hrs) 062810 3785 4.0 0.5 Pt ion treatment (Pt wires, 99.99%) Length Wire Volume Voltage Frequency Time of Wire Diameter (mL) (V) (Hz) (hrs) (in/cm) (mm) 800 130 60 0.5 2.01/5.1 1 Component 2-Gold Solution Pretreatment-Au GZA Run Volume NaHCO3 time ID (mL) (grams) (hrs) 061610 800 1.0 0.5 Au Nanoparticle treatment (Au wires, 99.99%) Length Wire Volume Voltage Frequency Time of Wire Diameter (mL) (V) (Hz) (hrs) (in/cm) (mm) 800 300 60 0.5 6.25/15.88 0.5 Mixture Comp. Comp. H2O2 H2O2 Run 1 Vol. 2 (Au) Concentration Vol ID (Pt) (mL) Vol. (mL) (v/v %) (μL) MT-55-04 300 700 0.800 250 - In general, this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 d and 11 b. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for this example, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. The precise electrical connections are described elsewhere herein.Control devices 20, as illustrated inFIGS. 8 c and 8 j were connected to theelectrodes 1/5 and 5/5, respectively. However, due to the relatively short run times in each “Run ID,” there was no need to actuate thecontrol devices 20. Thus, theends 9′ of theelectrodes trough member 30 b′. - The amount of potassium hydroxide (Fisher Scientific, Cat #P250-500) processing enhancer used in Run ID “PB-53” was about 0.604 grams/gallon (i.e., about 0.16 mg/mL.). The feed electrodes were platinum wires (1 mm/0.040″ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - The AC power source 501AC utilized a Chroma 61604 programmable unit. In particular, sine wave AC frequencies at 80 Hz were utilized to make suspensions of Pt ions and/or Pt colloids, in accordance with the teachings herein. The applied voltage was 215 volts with an applied current between about 4.0 amps and about 5.0 amps.
- The resulting Pt-water-based material was then allowed to cool to approximately 50 degrees Celsius. At that point the Pt-water-based material was fed into another separate and different trough unit as described below.
- In general, this additional trough which utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a. Electrical device 501AC, illustrated inFIG. 13 was used as the power supply for examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found elsewhere herein.Control devices 20, illustrated inFIGS. 8 c and 8 j were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected tocontrol devices 20 and 20 i which automatically adjusted the height of, for example, eachelectrode 5/5 in each electrode set 5/5; had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - In particular, a sine wave AC frequency at 60 Hz was utilized to form the bi-metallic nanocrystalline suspension in accordance with the teachings herein. The platinum-water based material “PB-53,” as discussed above, was fed as a raw material via
pump 40 intoplasma trough section 30 a′ as illustrated inFIG. 10 c . The AC power source 501AC utilized a Chroma 61604 programmable AC source. The applied voltage was about 260 volts for approximately two minutes followed by about 220 volts for the duration of the run. The applied current varied between about 4 amps and about 5 amps. - Transmission electron microscopy (TEM) was used to examine the bi-metallic nanocrystals made according to this Example. In particular, TEM sample preparation was identical to the methods described earlier in the High Resolution TEM & EDS Section. The TEM micrographs show that the formed bi-metallic nanocrystals exist in some instances in a chain-like form of gold nanocrystals with platinum interconnects as evident in
FIGS. 25 a and 25 b dried from suspension GPB-0001, made according to this Example. - The total amount of platinum species and gold species contained within this bi-metallic nanocrystalline suspension was about 1.6 ppm and 7.7 ppm, respectively, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 15 summarizes key processing parameters used in conjunction with
FIGS. 9 and 10 b. Table 15 also discloses: 1) resultant “ppm” (i.e., atomic platinum and gold concentrations.) -
TABLE 15 GPB-001/ Run ID: PB-53 PGT-001 Feed: PE/Concentration(mg/ml) KOH/0.00156 PB-53 Input Temp ° C. at 32 23 45 Output Temp ° C. at 32 71 79 Flow In (ml/min) 215 230 Rate: Out (ml/min) 180 200 Volts: Set # 1 750 750 Set #'s 2-8 215 260: 0-2 min/220 Set #'s 2-8 80 60 frequency, Hz Wire Diameter (mm) 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b Produced Pt/Au PPM 1.6/NA 1.6/7.7 Dimensions Plasma 4 Figs. 9 9 Process Figures 10a, 10d 10c, 11a M (in/mm) 1.5/38 1.5/38 L T (in/mm)36/914 36/914 d (in/mm) 1/25 1/25 S (in/mm) 1.5/38 1.5/38 Electrode Curr. (A) 0.63 0.69 Total Curr. Draw (A) 4.40 4.40 “c-c” (mm) 76 76 Set 1electrode # 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 electrode # 5a 5a “c-c” (mm) 102 102 Set 2electrode # 5b 5b “x” (in/mm) n/a n/ a electrode # 5b′ 5b′ “c-c” (mm) 76 76 Set 3electrode # 5c 5c electrode # 5c′ 5c′ “c-c” (mm) 76 76 Set 4electrode # 5d 5d electrode # 5d′ 5d′ “c-c” (mm) 127 127 Set 5electrode # 5e 5e electrode # 5e′ 5e′ “c-c” (mm) 127 127 Set 6electrode # 5f 5f electrode # 5f′ 5f′ “c-c” (mm) 152 152 Set 7electrode # 5g 5g electrode # 5g′ 5g′ “c-c” (mm) 178 178 Set 8electrode # 5h 5h electrode # 5h′ 5h′ “c-c” (mm) 76 76 - This Example utilized a batch process according to the present invention.
FIG. 12 a shows the apparatus used to condition theliquid 3. Once conditioned, theliquid 3′ was processed in the apparatus shown inFIG. 12 c or 12 d, for platinum ions/particles and bi-metallic nanocrystals, respectively. The overall process created a bi-metallic nanocrystal suspension, as described below and summarized in Table 16. - Initially, platinum ions and/or particles were prepared by the following process. Approximately 0.580 grams/gallon (i.e., about 0.153 mg/mL) of processing enhancer potassium hydroxide (i.e., KOH) was added to 1 gallon of de-ionized water. The amount of time that the
water 3 with processing enhancer was exposed to theplasma 4 was about 30 minutes, prior to subsequent processing in the apparatus shown inFIG. 24 c. - The applied voltage for the
plasma 4 made by theelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. Note that in Table 16 (and elsewhere herein) the reference to “GZA” is synonomous with creation ofplasma 4. - A second and different transformer was electrically connected to the
electrodes 5 a/5 b shown inFIG. 12 c . This transformer was an hy AC power source having a voltage range of 0-300V, a frequency range of 47-400 Hz and a maximum power rating of 1 kVA. The applied voltage was about 100 volts with a frequency of 60 hertz for about 3 hours of operation. The diameter of the platinum wire electrodes was about 1 mm. - Subsequently, the platinum species and water material prepared above was further processed as described below. The platinum species and water material was then processed via the apparatus in
FIG. 12 d with gold electrodes (99.99%, 0.5 mm) for about 3 hours, with an hy AC power source having an applied voltage of about 180 volts and about 47 hertz. The average radius of the bi-metallic nanocrystals produced was about 14.6 nm as measured by ViscoTek. The suspension contained about 7.3 ppm of Au and about 1.2 ppm of Pt, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. -
FIGS. 26 a and 26 b show representative TEM Photomicrographs and energy-dispersive x-ray spectra of the formed bi-metallic nanocrystals, respectively, dried from suspension ID #PGB002, made according to this Example 11. -
TABLE 16 Component 1Pretreatment-GZA Run Volume KOH time ID (mL) (grams) (hrs) Pt011011 3785 0.580 0.5 Pt ion treatment (Pt wires, 99.99%) Length Wire Volume Voltage Frequency Time of Wire Diameter (mL) (V) (Hz) (hrs) (in/cm) (mm) 3785 100 60 3 2.01/5.1 1 Component 2 N/A Run Volume NaHCO3 time ID (mL) (grams) (hrs) N/A N/A N/A N/A Composite Mix Mixture of Component 1 & 2Comp. Comp. Run 1 Vol. 2 Vol. Volume ID (mL) (mL) (mL) Pt011011 3785 0 3785 Gold Nanoparticle Treatment (Au wires, 99.99%) Length Wire Voltage Frequency Time Current of Wire Diameter (V) (Hz) (hrs) (A) (in/cm) (mm) 180 47 3.0 N/A 6.25/15.88 0.5 - In general, this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 d and 11 b. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for this Example, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively. However, due to the short run times in each “Run ID,” there was no need to actuate thecontrol devices 20. Accordingly, in reference toFIGS. 3 c and 9 c , theends 9′ of theelectrodes trough member 30 b′. This example utilized about 3.5 g/gallon (i.e., about 0.925 mg/mL) of NaHCO3 as a processing enhancer and a flow rate of about 150 ml/min. - In particular, sine wave AC frequencies at 5 Hz were utilized to make Pt species in water in accordance with the teachings herein. The function generator 501FG provided sine waves at frequencies less than 15 Hz to power supply 501AC, Chroma 61604 programmable AC source, which subsequently amplified the input signal to about 150V. The applied current varied between about 5.0 amps to about 6.5 amps.
- The amount of platinum species produced in the water was about 15.9 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 17 summarizes key processing parameters used in conjunction with
FIGS. 9 and 10 d. Table 17 also discloses resultant “ppm” (i.e., atomic platinum nanocrystal concentrations.) -
TABLE 17 Run ID: PB56001 Flow In (ml/min) 150 Rate: Out (ml/min) 140 Volts: Set # 1750 Set #'s 2-8 150 Set #'s 2-8 5 frequency, Hz PE: NaHCO3 (mg/ml) 0.92 Wire Diameter (mm) 1.0 Contact “WL” (in/mm) 1/25 Electrode Separation “y” (in/mm) .25/6.4 Electrode Config. Figure 8b Produced Pt PPM 15.9 Output Temp ° C. at 32 79 Dimensions Plasma 4 Figs. 9 Process Figures 10a, 10d M (in/mm) 1.5/38 L T (in/mm)36/914 d (in/mm) 1/25 S (in/mm) 1.5/38 Electrode Curr. (A) 0.92 Total Curr. Draw (A) 5.75 “c-c” (mm) 76 Set 1electrode # 1a “x” (in/mm) 0.25/6.4 electrode # 5a “c-c” (mm) 102 Set 2electrode # 5b “x” (in/mm) n/a electrode # 5b′ “c-c” (mm) 76 Set 3electrode # 5c electrode # 5c′ “c-c” (mm) 76 Set 4electrode # 5d electrode # 5d′ “c-c” (mm) 127 Set 5electrode # 5e electrode # 5e′ “c-c” (mm) 127 Set 6electrode # 5f electrode # 5f′ “c-c” (mm) 152 Set 7electrode # 5g electrode # 5g′ “c-c” (mm) 178 Set 8electrode # 5h electrode # 5h′ “c-c” (mm) 76 - In general, this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 d and 11 b. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for this Example, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments.Control devices 20, illustrated inFIGS. 8 c and 8 j were connected to theelectrodes 1/5 and 5/5, respectively. However, due to the short run times in each “Run ID,” there was no need to actuate thecontrol devices 20. Accordingly, theends 9′ of theelectrodes trough member 30 b′. This example utilized about 2.5 g/gallon (i.e, about 0.661 mg/mL) of NaHCO3 as a processing enhancer and a flow rate of about 220 ml/min. - In particular, sine wave AC frequencies at 5 Hz were utilized to make Pt species in water in accordance with the teachings herein. The function generator 501FG provided sine waves at frequencies less than 15 Hz to power supply 501AC, Chroma 61604 programmable AC source, which subsequently amplified the input signal to about 175V. The applied current varied between about 4.0 amps to about 6.5 amps.
- The amount of platinum species produced in the water suspensions was about 7.8 ppm, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 18 summarizes key processing parameters used in conjunction with
FIGS. 9 and 10 d. Table 18 also discloses resultant “ppm” (i.e., atomic platinum nanocrystal concentrations.) -
TABLE 18 Run ID: PB57001 Flow In (ml/min) 220 Rate: Out (ml/min) 200 Volts: Set # 1750 Set #'s 2-8 175 Set #'s 2-8 5 frequency, Hz PE: NaHCO3 (mg/ml) 0.66 Wire Diameter (mm) 1.0 Contact “WL” (in/mm) 1/25 Electrode Separation “y” (in/mm) .25/6.4 Electrode Config. Figure 8b Produced Pt PPM 7.8 Output Temp ° C. at 32 61 Dimensions Plasma 4 Figs. 9 Process Figures 10a, 10d M (in/mm) 1.5/38 L T (in/mm)36/914 d (in/mm) 1/25 S (in/mm) 1.5/38 Electrode Curr. (A) 0.61 Total Curr. Draw (A) 4.58 “c-c” (mm) 76 Set 1electrode # 1a “x” (in/mm) 0.25/6.4 electrode # 5a “c-c” (mm) 102 Set 2electrode # 5b “x” (in/mm) n/a electrode # 5b′ “c-c” (mm) 76 Set 3electrode # 5c electrode # 5c′ “c-c” (mm) 76 Set 4electrode # 5d electrode # 5d′ “c-c” (mm) 127 Set 5electrode # 5e electrode # 5e′ “c-c” (mm) 127 Set 6electrode # 5f electrode # 5f′ “c-c” (mm) 152 Set 7electrode # 5g electrode # 5g′ “c-c” (mm) 178 Set 8electrode # 5h electrode # 5h′ “c-c” (mm) 76 - In general, this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for the examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected to controldevices 20 and 20 i which automatically adjusted the height of, for example, each electrode or 5/5 in each electrode set 5/5; had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - The amount of potassium hydroxide (Fisher Scientific, Cat #P250-500) processing enhancer used in Run ID “PB-106-2” was about 0.450 grams/gallon (i.e., about 0.119 mg/mL). In addition, the amount of sodium bicarbonate (Fisher Scientific, Cat #S631-3) used in Run ID “PB-106-2” was about 0.850 grams/gallon (i.e., about 0.22 mg/mL). The feed electrodes were platinum wires (1 mm/0.040″ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - The AC power source 501AC utilized a Chroma 61604 programmable unit. In particular, sine wave AC frequencies at 80 Hz were utilized to make at least one platinum species in water in accordance with the teachings herein. The applied voltage was about 215 volts with an applied current between about 4.0 amps and about 7.0 amps.
- The resulting platinum species in water material was then allowed to cool overnight to approximately 23 degrees Celsius. At that point the Pt-water-based material was fed into a second separate and different trough unit as described below.
- In general, this second trough utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected to controldevices 20 and 20 i which automatically adjusted the height of, for example, eachelectrode 5/5 in each electrode set 5/5 had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - In particular, a sine wave AC frequency at 60 Hz was utilized to make a gold nanocrystal suspension or colloid or ion, in accordance with the teachings herein. The platinum-water based material “PB-106-2,” as discussed above, was fed via
pump 40 intoplasma trough section 30 a′ as illustrated inFIG. 10 c . The AC power source 501AC utilized a Chroma 61604 programmable AC source. The applied voltage was about 260 volts for approximately two minutes followed by about 220 volts for the duration of the run. The applied current varied between about 4 amps and about 7 amps. - The total amount of platinum and gold contained within the bi-metallic nanocrystal suspension this material was about 3.0 ppm and 9.2 ppm, respectively, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 19 summarizes key processing parameters used in conjunction with
FIGS. 9 and 10 b. Table 19 also discloses: 1) resultant “ppm” (i.e., atomic platinum and gold concentrations.) -
TABLE 19 Run ID PB-106-2 GPB-032 Process NaHCOO3 (mg/mL) 0.225 PB-106-2 Enhancer KOH (mg/mL) 0.119 Input Temp ° C. at 32 24 24 Output Temp ° C. at 32 86 84 Flow Rate In (ml/min) 190 200 Out (ml/min) 175 180 Volts: Set # 1750 750 Set #'s 2-8 215 260: 0-2 min/220 Set #'s 2-8 80 60 frequency, Hz Wire Diameter (mm) 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b Produced Au/Pt PPM NA/3.0 9.2/3.0 Hydrodynamic Radius (nm) N/A 15.39 Zeta Potential (mV) N/A −53.0 Dimensions Plasma 4 Figs. 9 9 Process Figures 10c, 11a 10c, 11a M (in/mm) 1.5/38 1.5/38 LT (in/mm) 36/914 36/914 d (in/mm) 1/25 1/25 S (in/mm) 1.5/38 1.5/38 Total Curr. Draw (A) 6.34 6.53 “c-c” (mm) 76 76 Set 1electrode # 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 electrode # 5a 5a “c-c” (mm) 102 102 Set 2Electrode Pair # 5b & 5b’ 5b & 5b’ “c-c” (mm) 76 76 Set 3Electrode Pair # 5c & 5c’ 5c & 5c’ “c-c” (mm) 76 76 Set 4Electrode Pair # 5d & 5d’ 5d & 5d’ “c-c” (mm) 127 127 Set 5Electrode Pair # 5e & 5e’ 5e & 5e’ “c-c” (mm) 127 127 Set 6Electrode Pair # 5f & 5f’ 5f & 5f’ “c-c” (mm) 152 152 Set 7Electrode Pair # 5g & 5g’ 5g & 5g’ “c-c” (mm) 178 178 Set 8Electrode Pair # 5h & 5h’ 5h & 5h’ “c-c” (mm) 76 76 - In this Example, a Zeta-Sizer “Nano-ZS” produced by Malvern Instruments was utilized to determine zeta potential (the specifics of which are described earlier herein). For each measurement a 1 ml sample was filled into clear disposable zeta cell DTS1060C. Dispersion Technology Software, version 5.10 was used to run the Zeta-Sizer and to calculate the zeta potential. The following settings were used: dispersant—water, temperature—25° C., viscosity—0.8872 cP, refraction index—1.330, dielectric constant—78.5, approximation model—Smoluchowski. Three replications of 60 runs per individual replicate were performed for each sample. Energy absorption spectra was obtained for this sample (GPB-032) using Uv-Vis spectroscopy methods as outlined elsewhere herein.
FIG. 27 contains the UV-Vis data collected for this sample (GPB-032), specifically displaying the 350-900 nm range. - In general, this example utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected to controldevices 20 and 20 i which automatically adjusted the height of, for example, eachelectrode 5/5 in each electrode set 5/5 had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - The the amount of sodium bicarbonate (Fisher Scientific, Cat #S631-3) used in Run ID “PB-74” was about 2.5 grams/gallon (i.e., about 0.66 g/L). The feed electrodes were platinum wires (1 mm/0.040″ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - The AC power source 501AC utilized a Chroma 61604 programmable unit. In particular, sine wave AC frequencies at 80 Hz were utilized to make at least one platinum species in water, in accordance with the teachings herein. The applied voltage was 175 volts with an applied current between about 4.0 amps and about 7.0 amps.
- The resulting platinum species in water material was then allowed to cool overnight to approximately 23 degrees Celsius. At that point the Pt-water-based material was fed into a second, separate and different trough unit as described below.
- In general, this second trough utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected to controldevices 20 and 20 i which automatically adjusted the height of, for example, eachelectrode 5/5 in each electrode set 5/5 had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - In particular, a sine wave AC frequency at 60 Hz was utilized to make a gold nanocrystal suspension or colloid or ion, in accordance with the teachings herein. The platinum-water based material “PB-74,” as discussed above, was fed via
pump 40 intoplasma trough section 30 a′ as illustrated inFIG. 10 b . The AC power source 501AC utilized a Chroma 61604 programmable AC source. The applied voltage was initially set to 200 volts but was set to 165 volts due to the initial current reading falling out of the normal range, typically between 2.5 A-3.5 A. The applied current varied between about 4 amps and about 7 amps. - The total amount of atomic platinum and gold contained within the bi-metallic nanocrystal suspension was about 1.7 ppm and 7.8 ppm, respectively, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein. It should be noted that this particular Au—Pt bi-metallic nanocrystal suspension was not stable as it settled over a period of time no later than four months after production. Accordingly, under certain sets of processing conditions, sodim bicarbonate by itself, without the addition of KOH or other suitable processing enhancers does not promote the development of highly stable Au—Pt bi-metallic nanocrystal suspensions. However, these suspensions could be suitable for some purposes.
- Table 20 summarizes key processing parameters used in conjunction with
FIGS. 9 and 10 b. Table 20 also discloses: 1) resultant “ppm” (i.e., atomic platinum and gold concentrations.) and 2) “Hydrodynamic Radius” (nm). -
TABLE 20 Run ID PB-74 GPB-010 Process NaHCOO3 (mg/mL) 0.661 PB-74 Enhancer Input Temp ° C. at 32 24 24 Output Temp ° C. at 32 70 64 Flow Rate In (ml/min) 190 200 Volts: Set # 1750 750 Set #'s 2-8 175 165 Set #'s 2-8 80 60 frequency, Hz Wire Diameter (mm) 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b Produced Au/Pt PPM NA/1.7 7.8/1.7 Hydrodynamic Radius (nm) N/A 115 Dimensions Plasma 4 Figs. 9 9 Process Figures 10a, 10d 10c, 11a M (in/mm) 1.5/38 1.5/38 LT (in/mm) 36/914 36/914 d (in/mm) 1/25 1/25 S (in/mm) 1.5/38 1.5/38 Total Curr. Draw (A) 5.16 4.67 “c-c” (mm) 76 76 Set 1electrode # 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 electrode # 5a 5a “c-c” (mm) 102 102 Set 2Electrode Pair # 5b & 5b’ 5b & 5b’ “c-c” (mm) 76 76 Set 3Electrode Pair # 5c & 5c’ 5c & 5c’ “c-c” (mm) 76 76 Set 4Electrode Pair # 5d & 5d’ 5d & 5d’ “c-c” (mm) 127 127 Set 5Electrode Pair # 5e & 5e’ 5e & 5e’ “c-c” (mm) 127 127 Set 6Electrode Pair # 5f & 5f’ 5f & 5f’ “c-c” (mm) 152 152 Set 7Electrode Pair # 5g & 5g’ 5g & 5g’ “c-c” (mm) 178 178 Set 8Electrode Pair # 5h & 5h’ 5h & 5h’ “c-c” (mm) 76 76 - In general, this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected to controldevices 20 and 20 i which automatically adjusted the height of, for example, eachelectrode 5/5 in each electrode set 5/5 had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - The amount of potassium hydroxide (Fisher Scientific, Cat #P250-500) processing enhancer used in Run IDs “PB-83, 85, 87, and 88” was about 0.450 grams/gallon (i.e., about 0.12 mg/mL.). In addition, the amount of sodium bicarbonate (Fisher Scientific, Cat #S631-3) used in Run IDs “PB-83, 85, 87, and 88” was about 0.850 grams/gallon (i.e., about 0.22 mg/mL). The feed electrodes were platinum wires (1 mm/0.040″ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - The AC power source 501AC utilized a Chroma 61604 programmable unit. In particular, sine wave AC frequencies at 80 Hz were utilized to at least one platinum species in water in accordance with the teachings herein. The applied voltage was about 215 volts with an applied current between about 4.0 amps and about 7.0 amps.
- The resulting platinum species in water material was then allowed to cool overnight to approximately 23 degrees Celsius. At that point the Pt-water-based material was fed into a second, separate and different trough unit as described below.
- In general, this second trough utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected to controldevices 20 and 20 i which automatically adjusted the height of, for example, eachelectrode 5/5 in each electrode set 5/5 had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - In particular, a sine wave AC frequency at 5 Hz-200 Hz was utilized to make gold nanocrystal suspensions or colloids or ions, in accordance with the teachings herein. The platinum-water based material “PB-83, 85, 87, and 88,” as discussed above, was fed via
pump 40 intoplasma trough section 30 a′ as illustrated inFIG. 10 b . The AC power source 501AC utilized a Chroma 61604 programmable AC source. The applied voltage was about 260 volts for approximately two minutes followed by about 220 volts for the duration of the run. The applied current varied between about 4 amps and about 7 amps. - The total amount of atomic platinum and gold contained within the bi-metallic nanocrystal suspension are outlined in Tables 21a, 21b and 21c. Table 21a outlines the platinum run conditions used to form the platinum species in water and Tables 21b and 21c outline the run conditions used to form the Au—Pt bi-metallic nanocrystal suspensions.
- Table 21a summarizes key processing parameters used in conjunction with
FIGS. 9 and 10 c. Tables 21a, 21b and 21c also disclose: 1) Resultant “ppm” (i.e., atomic platinum and gold concentrations), 2) Hydrodynamic radius, and 3) Zeta Potential. - Energy absorption spectra was obtained for these samples (PGT024, PGT025, PGT026) using Uv-Vis spectroscopy methods as outlined elsewhere herein.
FIG. 28 a contains the UV-Vis data collected for thes samples (PGT024, PGT025, PGT026), specifically displaying the 350-900 nm range. - Energy absorption spectra was obtained for these samples (GPB-017, GPB-018, GPB-019, GPB-020, GPB-023) using Uv-Vis spectroscopy methods as outlined elsewhere herein.
FIG. 28 b contains the UV-Vis data collected for these samples (GPB-017, GPB-018, GPB-019, GPB-020, GPB-023), specifically displaying the 350-900 nm range. - A variety of Au—Pt bi-metallic nanocrystal suspensions were prepared at frequencies, as described in this Example, between the range of about 5 Hz-200 Hz. A representative comparison of particle size versus frequency is illustrated in
FIG. 28 c . -
TABLE 21a Run ID PB-83 PB-85 PB-87 PB-88 Process NaHCOO3 (mg/mL) 0.225 0.225 0.225 0.225 Enhancer KOH (mg/mL) 0.119 0.119 0.119 0.119 Input Temp ° C. at 32 23 25 25 24 Output Temp ° C. at 32 74 80 81 76 Flow Rate In (ml/min) 220 220 220 220 Volts: Set # 1750 750 750 750 Set #'s 2-8 215 215 215 215 Set #'s 2-8 80 80 80 80 frequency, Hz Wire Diameter (mm) 1.0 1.0 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b 8b 8b Produced Pt PPM 1.9 2.2 2.3 2.1 Dimensions Plasma 4 Figs. 9 9 9 9 Process Figures 10a, 10d 10a, 10d 10a, 10d 10a, 10d M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 LT (in/mm) 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 S (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 Total Curr. Draw (A) 5.12 5.52 5.87 5.45 “c-c” (mm) 76 76 76 76 Set 1 electrode # 1a 1a 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 electrode # 5a 5a 5a 5a “c-c” (mm) 102 102 102 102 Set 2 Electrode Pair # 5b & 5b’ 5b & 5b’ 5b & 5b’ 5b & 5b’ “c-c” (mm) 76 76 76 76 Set 3 Electrode Pair # 5c & 5c’ 5c & 5c’ 5c & 5c’ 5c & 5c’ “c-c” (mm) 76 76 76 76 Set 4 Electrode Pair # 5d & 5d’ 5d & 5d’ 5d & 5d’ 5d & 5d’ “c-c” (mm) 127 127 127 127 Set 5 Electrode Pair # 5e & 5e’ 5e & 5e’ 5e & 5e’ 5e & 5e’ “c-c” (mm) 127 127 127 127 Set 6 Electrode Pair # 5f & 5f’ 5f & 5f’ 5f & 5f’ 5f & 5f’ “c-c” (mm) 152 152 152 152 Set 7 Electrode Pair # 5g & 5g’ 5g & 5g’ 5g & 5g’ 5g & 5g’ “c-c” (mm) 178 178 178 178 Set 8 Electrode Pair # 5h & 5h’ 5h & 5h’ 5h & 5h’ 5h & 5h’ “c-c” (mm) 76 76 76 76 -
TABLE 21b Run ID GPB-017 GPB-018 GPB-019 GPB-020 GPB-021 Process NaHCOO3 (mg/mL) PB-83 PB-83 PB-83 PB-85 PB-85 Enhancer KOH (mg/mL) Input Temp ° C. at 32 25 25 25 27 27 Output Temp ° C. at 32 79 78 78 81 83 Flow Rate In (ml/min) 230 230 230 230 230 Volts: Set # 1750 750 750 750 750 Set #'s 2-8 220 220 220 260 V: 220 0-2 min/220 Set #'s 2-8 frequency, Hz 20 40 80 5 10 Wire Diameter (mm) 1.0 1.0 1.0 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 1/25 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 .25/6.4 .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b 8b 8b 8b Produced Au/Pt PPM 3.1/2.0 5.8/2.0 10.5/2.0 1.1/2.3 1.7/2.3 Hydrodynamic Radius (nm) 18.96 16.59 20.58 24.96 51 Zeta Potential (mV) −39.0 −38.0 −42.0 −45.0 −38.0 Dimensions Plasma 4 Figs. 9 9 9 9 9 Process Figures 10c, 11a 10c, 11a 10c, 11a 10c, 11a 10c, 11a M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 LT (in/mm) 36/914 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 1/25 S (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 1.5/38 Total Curr. Draw (A) 5.84 5.82 5.81 5.66 5.82 “c-c” (mm) 76 76 76 76 76 Set 1 electrode # 1a 1a 1a 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 electrode # 5a 5a 5a 5a 5a “c-c” (mm) 102 102 102 102 102 Set 2 Electrode Pair # 5b & 5b’ 5b & 5b’ 5b & 5b’ 5b & 5b’ 5b & 5b’ “c-c” (mm) 76 76 76 76 76 Set 3 Electrode Pair # 5c & 5c’ 5c & 5c’ 5c & 5c’ 5c & 5c’ 5c & 5c’ “c-c” (mm) 76 76 76 76 76 Set 4 Electrode Pair # 5d & 5d’ 5d & 5d’ 5d & 5d’ 5d & 5d’ 5d & 5d’ “c-c” (mm) 127 127 127 127 127 Set 5 Electrode Pair # 5e & 5e’ 5e & 5e’ 5e & 5e’ 5e & 5e’ 5e & 5e’ “c-c” (mm) 127 127 127 127 127 Set 6 Electrode Pair # 5f & 5f’ 5f & 5f’ 5f & 5f’ 5f & 5f’ 5f & 5f’ “c-c” (mm) 152 152 152 152 152 Set 7 Electrode Pair # 5g & 5g’ 5g & 5g’ 5g & 5g’ 5g & 5g’ 5g & 5g’ “c-c” (mm) 178 178 178 178 178 Set 8 Electrode Pair # 5h & 5h’ 5h & 5h’ 5h & 5h’ 5h & 5h’ 5h & 5h’ “c-c” (mm) 76 76 76 76 76 -
TABLE 21c Run ID GPB-023 PGT024 PGT025 PGT026 Process NaHCOO3 (mg/mL) PB-85 PB-87 PB-83 PB-85 Enhancer KOH (mg/mL) Input Temp ° C. at 32 27 27 25 25 Output Temp ° C. at 32 83 83 84 83 Flow Rate In (ml/min) 230 230 230 230 Volts: Set # 1750 750 750 750 Set #'s 2-8 220 260 V: 0-2 260 V: 0-2 220 min/220 min/220 Set #'s 2-8 200 60 30 100 frequency, Hz Wire Diameter (mm) 1.0 1.0 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b 8b 8b Produced Au/Pt PPM 12.3/2.3 8.5/2.7 4.8/2.6 12.2/2.5 Hydrodynamic Radius (nm) 41.31 19.17 17.43 28.84 Zeta Potential (mV) −44.0 −40.0 −56.0 −50.0 Dimensions Plasma 4 Figs. 9 9 9 9 Process Figures 10c, 11a 10c, 11a 10c, 11a 10c, 11a M (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 LT (in/mm) 36/914 36/914 36/914 36/914 d (in/mm) 1/25 1/25 1/25 1/25 S (in/mm) 1.5/38 1.5/38 1.5/38 1.5/38 Total Curr. Draw (A) 6.04 5.81 5.86 5.82 “c-c” (mm) 76 76 76 76 Set 1 electrode # 1a 1a 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 0.25/6.4 0.25/6.4 electrode # 5a 5a 5a 5a “c-c” (mm) 102 102 102 102 Set 2 Electrode Pair # 5b & 5b’ 5b & 5b’ 5b & 5b’ 5b & 5b’ “c-c” (mm) 76 76 76 76 Set 3 Electrode Pair # 5c & 5c’ 5c & 5c’ 5c & 5c’ 5c & 5c’ “c-c” (mm) 76 76 76 76 Set 4 Electrode Pair # 5d & 5d’ 5d & 5d’ 5d & 5d’ 5d & 5d’ “c-c” (mm) 127 127 127 127 Set 5 Electrode Pair # 5e & 5e’ 5e & 5e’ 5e & 5e’ 5e & 5e’ “c-c” (mm) 127 127 127 127 Set 6 Electrode Pair # 5f & 5f’ 5f & 5f’ 5f & 5f’ 5f & 5f’ “c-c” (mm) 152 152 152 152 Set 7 Electrode Pair # 5g & 5g’ 5g & 5g’ 5g & 5g’ 5g & 5g’ “c-c” (mm) 178 178 178 178 Set 8 Electrode Pair # 5h & 5h’ 5h & 5h’ 5h & 5h’ 5h & 5h’ “c-c” (mm) 76 76 76 76 - In general, this Example utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a to make Au—Pt bi-metallic nanocrystal suspensions. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.Control devices 20, illustrated inFIGS. 18 c and 18 j , were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected to controldevices 20 and 20 i which automatically adjusted the height of, for example, eachelectrode 5/5 in each electrode set 5/5 had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - The amount of potassium hydroxide (Fisher Scientific, Cat #P250-500) processing enhancer used in Run ID “PB-118” was about 0.450 grams/gallon (i.e., about 0.12 mg/mL.). In addition, the amount of sodium bicarbonate (Fisher Scientific, Cat #S631-3) used in Run ID “PB-118” was about 0.850 grams/gallon (i.e., about 0.22 mg/mL). The feed electrodes were platinum wires (1 mm/0.040″ dia.), 99.99%, obtained from Hi-Rel Alloys LTD (Ontario, Canada.)
- The applied voltage for each
plasma 4 made byelectrode 1 was about 750 volts. This voltage was achieved by a transformer 60 (i.e., the Balanced Mid-Point Referenced Design) discussed elsewhere herein. - The AC power source 501AC utilized a Chroma 61604 programmable unit. In particular, sine wave AC frequencies at 80 Hz were utilized to make at least one platinum species in water, in accordance with the teachings herein. The applied voltage was about 215 volts with an applied current between about 4.0 amps and about 7.0 amps.
- The resulting platinum species in water material was then allowed to cool overnight to approximately 23 degrees Celsius. At that point the Pt-water-based material was fed into a second, separate and different trough unit as described below.
- In general, this second trough utilized certain embodiments of the invention associated with the apparatuses generally shown in
FIGS. 9, 10 c and 11 a. Electrical device 501AC, illustrated inFIG. 13 , was used as the power supply for examples contained herein, while function generator 501FG was sometimes used to drive 501AC. This transformer was an AC power source (Chroma 61604) having an AC voltage range of 0-300V, a frequency range of 15-1000 Hz and a maximum power rating of 2 kVA. Electrical connectivity discussions can be found in the detailed description of the preferred embodiments section.Control devices 20, illustrated inFIGS. 8 c and 8 j , were connected to theelectrodes 1/5 and 5/5, respectively, andelectrodes 5/5 were actuated at a rate of about 1″ per 8 hours. The eightelectrode sets 1/5 and 5/5 were all connected to controldevices 20 and 20 i which automatically adjusted the height of, for example, eachelectrode 5/5 in each electrode set 5/5 had 2 female receiver tubes o5 a/o5 a′-o5 g/o5 g′ which were connected to a bottom portion of thetrough member 30 b′ such that the electrodes in each electrode set 5/5 could be removably inserted into each female receiver tube o5 when, and if, desired. - In particular, a sine wave AC frequency at 60 Hz was utilized to make a gold nanocrystal suspension or colloid or ion, in accordance with the teachings herein. The platinum-water based material “PB-118,” as discussed above, was fed via
pump 40 intoplasma trough section 30 a′ as illustrated inFIG. 10 c . The AC power source 501AC utilized a Chroma 61604 programmable AC source. The applied voltage was about 260 volts for approximately two minutes followed by about 220 volts for the duration of the run. The applied current varied between about 4 amps and about 7 amps. - The total amount of atomic platinum and gold contained within the bi-metallic nanocrystalline suspension was about 3.2 ppm and 9.3 ppm, respectively, as measured by the atomic absorption spectroscopy techniques discussed elsewhere herein.
- Table 23 summarizes key processing parameters used in conjunction with
FIGS. 9 and 11 a. Table 23 also discloses: 1) resultant “ppm” (i.e., atomic platinum and gold concentrations.), 2) “Hydrodynamic Radius” and 3) “Zeta Potential.” - High-resolution transmission electron microscopy (HRTEM) was performed using a Philips CM300 FEG High Resolution Transmission Electron Microscope described elsewhere herein. Scanning transmission electron microscopy (STEM) was also performed on the CM300 in STEM mode. Calibration was performed prior to analysis via an internal calibration procedure within the instrument computer.
FIGS. 29 a and 29 c are representative TEM micrographs.FIGS. 29 b and 29 d are representative EDS spectra of dried nanocrystals inFIGS. 29 a and 29 c .FIGS. 29 e, 29 f and 29 g are STEM mappings of dried Au—Pt bi-metallic nanocrystals dried from the nanocrystal suspensions. - Energy absorption spectra were obtained for this sample (GPB-040) using Uv-Vis spectroscopy methods as outlined elsewhere herein.
FIG. 30 contains the UV-Vis data collected for this sample (GPB-040), specifically displaying the 350-900 nm range. - GPB-040 concentrated samples were prepared via Tangential Flow Filtration (TFF), as described herein where the diafiltration buffer was substituted with de-ionized water to remove the process enhancer from the solution. GPB-040 was concentrated 20 fold by volume three times, each time reconstituting with de-ionized water. Subsequently, TFF concentrated GPB-040 was then centrifuged at 11,000 rpm for 10 minutes resulting in the presence of a Au—Pt bi-metallic pellet at the bottom of a 1.5 mL centrifuge tube. Approximately 24 tubes were used to collect a final sample of about 1.5 mL with a concentration that is about 400 times greater than the starting solution. This solution was then deposited onto the sample stub as discussed below.
- In order to concentrate the bi-metallic nanocrystals in GPB-040, a tangential flow filtration (TFF) process was utilized. In the process filtration is a pressure driven separation process that uses membranes to separate nanocrystals in the suspension based on their size and/or charge differences. In TFF, the fluid is pumped tangentially along the surface of the membrane. A schematic of a simple TFF system is shown in
FIG. 31 c. - A
feed tank 1001 provides fluid to afeed pump 1002 and into afiltration module 1003. The filtrate stream 1004 is discarded. Retentate is diverted through theretentate valve 1005 and returned as 1006 into thefeed tank 1001. During each pass of the fluid over the surface of the membrane in thefiltration module 1003, the applied pressure forces a portion of the fluid through the membrane and into the filtrate stream, 1004. Any particulates and macromolecules that are too large to pass through the membrane pores are retained on the upper stream and swept along by the tangential flow into the retentate, 1006. The retentate, having a higher concentration of colloidal particles, is returned back to the feed tank, 1001. If there is no diafiltration buffer added to the feed tank, then the colloid volume in the feed tank, 1001, is reduced by the amount of filtrate removed and the suspension becomes concentrated. - In this example, Millipore Pellicon XL cassettes were used with 5 kDa and 10 kDa MWCO cellulose membranes. The retentate pressure was set to 40 PSI by a retentate valve, 1005. 10 kDa membrane allows approximately 4 times higher filtrate flow rate related to a 5 kDa membrane under the same transmembrane pressure, which is expected for a larger pore size. At the same time, pores of 10 kDa membrane are small enough to retain all formed bi-metallic nanocrystals in the retentate in GPB-040.
- Surface chemical analysis of bi-metallic gold-platinum nanocrystals was performed by X-ray photoelectron spectroscopy (XPS.) The spectra were collected using a Physical Electronics (PHI) Model 5400 photoelectron spectrometer equipped with a Mg K-alpha source operating at 300 W beam power with an accelerating voltage of 15 kV. Ejected photoelectrons were detected by a hemispherical analyzer that provided both high sensitivity and resolution. The operating pressure in the sampling chamber was below 5×10−8 Torr during analysis.
- Spectra were collected within two ranges, (i.e., a low resolution survey scan and a higher resolution multiplex scan in specific regions of interest). Survey scans were taken between binding energies of 0-1200 eV while higher resolution scans were taken between 80-100 eV and 65-85 eV. Elemental gold exhibits a multiplet (4f512 & 4f712) at 87.6 eV and 83.9 eV, respectively, and information such as oxide composition and concentration can be determined from the expanded region at 80-100 eV. Platinum exhibits a multiplet (4f512 & 4f712) at 74.5 eV and 71.2 eV, respectively, and information such as concentration and oxide content can be determined from the expanded region at 65-85 eV.
- Sputter cleaning and depth profiling were carried out with a Sputter Ion Gun, (PHI, Model 04-303). The incident ion gun was operated at an accelerating voltage of 4.0 keV, and sample currents were maintained at about 25 mA across the sample area. The pressure in the main chamber was maintained at about 5×10−8 Torr. The corresponding raster size is 4×4 mm with a pressure of 25 mPa. Sputtering was done at intervals of 5, 10, 20, 30, 40, 50, 70, 90, 120, 180, & 240 minutes.
-
FIGS. 29 h-29 i are spectra collected from GPB-040, a gold-platinum bi-metallic nanocrystal suspension. The spectra were prepared by placing 100-200 uL of sample onto the sample stub and subsequently pulling a vacuum to dry the material onto the carbon tape. The chamber was then opened and another 100-200 uL was deposited. This process was repeated eleven times to produce a thin film of material on the carbon tape. - The initial survey scan,
FIG. 29 h , is useful in determining surface contaminants and elemental composition of the nanocrystals. Clearly labeled are peaks indicative of carbon, oxygen, platinum, and gold. The small carbon peak at 285 eV is from incomplete sample coverage of the carbon tape while the oxygen peak at 531 eV is likely a result of trapped oxygen due to the sample preparation technique; however in a layer of adsorbed oxygen may have become trapped in between drop depositions. Peaks at 690 eV and 750 eV can be attributed to fluorine sample chamber contamination and oxygen, respectively. In both instances the peaks disappeared after a 30 minute sputter. - Higher resolution multiplex scans,
FIG. 29 i , between 60 eV-100 eV provide additional information on the gold and platinum composition of the nanocrystals. TheAu 4f512 peak at 88 eV contains a small shoulder that can be attributed to sample charging. After a 30 minute sputter, the flow of positive argon ions neutralized the sample and the shoulder disappeared. In addition, thePt 4f7/2 peak rises after the 30 minute sputter at about 71 eV. - As shown clearly in
FIGS. 29 a-g , Au—Pt bi-metallic nanocrystal solutions are heterogeneous in structure with respect to atomic platinum and atomic gold. As indicated by specific areas of interest inFIGS. 29 a and 29 c , energy dispersive spectra (EDS) were collected by condensing the electron beam of the TEM onto individual nanocrystals. Resultant EDS data is displayed inFIGS. 29 b and 29 d . In both cases, a platinum peak at about 9.4 keV and a gold peak at about 9.7 keV are present.FIGS. 29 e-g are Scanning Transmission Electron Microscopy (STEM) images of bi-metallic nanocrystals from suspension GPB-040.FIG. 29 e is a STEM image of at least four Au—Pt bi-metallic nanocrystals dried on a copper grid.FIGS. 29 f and g are platinum and gold EDS mappings, respectively, of the nanocrystals imaged inFIG. 29 e . It is clear fromFIGS. 29 f and 29 g that both platinum and gold exist heterogeneously throughout the examined nanocrystals. In addition,FIGS. 29 h and 29 i provide further evidence that the nanocrystal surfaces are both free from organic contamination and do not exhibit a core-shell behavior. The relative intensities of theAu 4f7/2 andPt 4f7/2 do not change as a function of sputtering time. One would expect the relative intensities of Pt to decrease if the nanocrystals were core-shell in nature. By combining both HRTEM, EDS, and XPS data, it is clear that the nanocrystals prepared by the methods disclosed in this Example are Au—Pt bi-metallic alloys. -
TABLE 23 Run ID PB-118 GPB-040 Process NaHCOO3 (mg/mL) 0.225 PB-120 Enhancer KOH (mg/mL) 0.119 Input Temp ° C. at 32 24 24 Output Temp ° C. at 32 88 86 Flow Rate In (ml/min) 190 200 Volts: Set # 1750 750 Set #'s 2-8 215 260: 0-2 min/220 Set #'s 2-8 80 60 frequency, Hz Wire Diameter (mm) 1.0 1.0 Contact “WL” (in/mm) 1/25 1/25 Electrode Separation “y” (in/mm) .25/6.4 .25/6.4 Electrode Config. Figure 8b 8b Produced Pt PPM 3.2 N/A Produced Au PPM N/A 9.3 Hydrodynamic Radius (nm) N/A 14.16 Zeta Potential (mV) N/A −47.0 Dimensions Plasma 4 Figs. 9 9 Process Figures 10c, 11a 10c, 11a M (in/mm) 1.5/38 1.5/38 LT (in/mm) 36/914 36/914 d (in/mm) 1/25 1/25 S (in/mm) 1.5/38 1.5/38 Total Curr. Draw (A) 6.25 6.04 “c-c” (mm) 76 76 Set 1electrode # 1a 1a “x” (in/mm) 0.25/6.4 0.25/6.4 electrode # 5a 5a “c-c” (mm) 102 102 Set 2Electrode Pair # 5b & 5b’ 5b & 5b’ “c-c” (mm) 76 76 Set 3Electrode Pair # 5c & 5c’ 5c & 5c’ “c-c” (mm) 76 76 Set 4Electrode Pair # 5d & 5d’ 5d & 5d’ “c-c” (mm) 127 127 Set 5Electrode Pair # 5e & 5e’ 5e & 5e’ “c-c” (mm) 127 127 Set 6Electrode Pair # 5f & 5f’ 5f & 5f’ “c-c” (mm) 152 152 Set 7Electrode Pair # 5g & 5g’ 5g & 5g’ “c-c” (mm) 178 178 Set 8Electrode Pair # 5h & 5h’ 5h & 5h’ “c-c” (mm) 76 76 - A dialysis bag technique permits the gradual concentration of colloids made according to the teachings herein. Colloidal suspensions were placed inside of a dialysis bag and the bag itself was immersed into an aqueous solution of a PEG-based polymer, which creates a negative osmotic pressure. The negative osmotic pressure resulted in the extraction of water from the colloid maintained within (i.e., inside) the dialysis bag.
- Specifically,
FIG. 31 a shows adialysis bag 2000, containing arepresentative colloid suspensions 3000. A suitable plastic container 5000 (made of HDPE plastic) and a PEG-basedpolymer material 1000 therein. - The dialysis membrane, which forms the
dialysis bag 2000, is characterized by molecular weight cut off (MWCO)—an approximate achieved threshold size above which larger-sized species will be retained inside of the membrane. Dialysis concentration was achieved by using a cellulose membrane having a 3.5 kDa MWCO for thedialysis bag 2000 and thepolymer solution 1000 was made from a PEG-8000 polymer. Under these conditions, water molecules and small ions could pass through the dialysis membrane of thebag 2000, but colloidal nanoparticles larger than the 3.5 kDa MWCO would be retained inside the dialysis bag. However, PEG-8000 molecules cannot pass through (i.e., due to their size) the membrane and remained outside of thedialysis bag 2000. -
FIG. 31 b shows that thedialysis bag 2000 shrank in volume (over time) relative to its size inFIG. 31 a . Thedialysis bag 2000 should not be allowed to collapse as liquid is removed from the bag. In this regard, nanocrystals that may remain on the inner surface of the bag should not be over-stressed so as to prevent their possible aggregation. - Each
dialysis bag 2000 was filled with approximately 400 to 500 mL ofnanocrystal suspension 3000, and maintained in the PEG-8000solution 1000 until the bag volume was reduced approximately 10 times in size and volume. Further suspension concentration, if required, occurred by combining 10× concentrated colloids from several bags into one bag and repeating the same set of concentration steps again.Dialysis bags 2000 can safely be used about 10 times without achieving any noticeable membrane fouling. - The starting PEG-8000
concentration 1000 in the polymer solution outside thedialysis bag 2000 was about 250 g/L and was naturally lowered in concentration due to water being drawn out from the colloid 3000 through the dialysis bags 2000 (i.e., due to the created osmotic pressure). Higher polymer concentrations and gentle stirring can increase the rate of water removal from the colloid 3000. - This dialysis process concentrated the gold colloids with no visible staining of the
dialysis bags 2000. The concentration of remaining gold nanocrystals insuspension 4000 was estimated by volume reduction and also measured by ICP-MS techniques (discussed in detail later herein). The remaining gold in thesuspension 4000 was similar to the gold concentration measured directly by ICP-MS techniques. However, in the case of the bi-metallic gold/platinum nanocrystal suspension, part of the platinum produced in the first electrochemical step was ionic, and some amount of this ionic form of platinum removal after the second electrochemical processing steps and passed through thedialysis bag 2000 during concentration. This effect resulted in a lower concentration factor for atomic platinum relative to atomic gold (all of the atomic gold was apparently in metallic form). In addition, the Au—Pt bi-metallic nanocrystal suspension slightly stained the membrane of thedialysis bag 2000 to a yellowish-green uniform color. - The dialysis bag technique was used to achieve a series of concentration ranges of two different colloidal suspensions that were used in a subsequent in-vitro cellular culture experiment. Specifically, Table 24 sets forth 9 different concentrations of metals in a formed gold suspension (NE10214) and in an Au/Pt bi-metallic suspension (GPB-032) the formations of which are described earlier herein. Concentration values were measured by inductively coupled plasma-mass spectrometry (ICP-MS) as desribed immediately below.
- The ICP-MS values were obtained from an Agilent 7700x
- The technique of inductively coupled plasma spectroscopy-mass spectrometry requires a liquid sample to be introduced into a sample chamber via a nebulizer, thus removing the larger droplets, and introducing a fine aerosol spray into the torch chamber carried via a supply of inert Argon gas. The torch temperature ranges between 8000K-10000K. The aerosol is instantly desolvated and ionized within the plasma and extracted into the first vacuum stage via the sampling cone and then subsequently passes through a second orifice, the skimmer cone. The ions are then collimated by the lens system and then focused by the ion optics.
The ion lenses allow the ICP-MS to achieve high signal sensitivity by preventing photons and neutral species from reaching the detector by mounting the quadrupole and detector off axis from the entering ion beam. The cell gas, Helium, is introduced into the ORS which is an octopole ion guide positioned between the ion lens assembly and the quadrupole. Interferences such as polyatomic species are removed via kinetic energy discrimination. The ions that pass through then proceed into the quadrupole mass analyzer which consists of four long metal rods. RF and DC voltages are applied at the rods and it is the variation in voltages that allow the rods to filter ions of specific mass-to-charge ratios.
The ions are then measured by the pulse analog detector. When an ion enters the electron multiplier, it strikes a dynode and creates an abundance of free electrons which then strike the next dynode, resulting in the creation of additional electrons. The amount of ions from a specific element correlates to the amount of electrons generated, thus resulting in more or less counts, or CPS. - Samples were prepared by diluting 500 μL of sample in 4.5 mL of 5% HNO3/2% HCl for 30 minutes at 70° C. Samples were prepared in triplicate. Subsequently, samples were transferred to a polypropylene test tube which was then placed in a rack in the Cetac autosampler.
- The Agilent ICP-MS 7700x plasma was turned on and a start up procedure was initialized. The plasma was allowed to warm up for 26 minutes prior to running the initial optimization. After successful completion of the optimization steps, the instrument was then ready for analysis. A quick manual tune was performed and the signal of low, mid, and high masses (59, 89, & 205) were checked to ensure that the instrument was within our internal specifications. Afterwards, the internal standard line tubing was switched from a 5% HNO3 blank to an internal standard solution containing In 115.
- Calibration samples and independent continuous concentration verification (ICCV) standards were prepared from external stock solutions prepared by SPEX CertiPrep. Multi-Element 3 calibration standards containing gold were serially diluted from 10 ppm to 1000 ppb, 100 ppb, 10 ppb, and 1 ppb, respectively. A blank solution of the diluent, 5% HNO3/2% HCl, was used as the 0 ppb standard. The ICCV sample was placed in a sample vial and placed on a rack with the calibration standards.
Prior to sample analysis, a calibration curve was created by measuring 0 ppb, 1 ppb, 10 ppb, 100 ppb, & 1000 ppb. Samples of interest were then measured with a 90 second 5% HNO3 rinse step in between sample uptake. After every 6 samples, the ICCV was run to ensure that the calibration curve was within 10% of the actual values. - Data was exported from the Mass-hunter Data analysis software to excel to be formatted and checked. Replicates were averaged together to obtain a mean concentration, standard deviation and relative standard deviation.
-
TABLE 24 NE10214 GPB-032 Au Au/Pt [Au], volume, Au + Pt volume, ID: ppm mL ID: ppm mL 1-1 981 10 2-1 982 3.2 1-2 800 10 2-2 800 3.5 1-3 600 10 2-3 600 4 1-4 400 10 2-4 400 4 1-5 200 10 2-5 385 5.2 1-6 80 10 2-6 180 4.5 1-7 40 10 2-7 40 4 1-8 20 10 2-8 20 4 1-9 8 10 2-9 8 4 1-10 blank 10 2-10 blank 4 control control - A cell line panel was assembled with 30 different human tumor types selected from the ATCC and DSMZ (all DSMZ cell lines are marked with “*”) culture banks and included typical bladder, breast, cervix, CNS, colon, H&N, lung, ovary, prostate, stomach, thyroid, uterus and vulva cancers. The 30 specific cell lines and tumor types are set forth in Table 25.
-
TABLE 25 CAT # Cell Line Morphology Cancer Type Organ ACC 414 647-V Epithelial Bladder Bladder ** ACC 279 BHT-101 Epithelial Endocrine Thyroid ** HTB-20 BT474 Epithelial Breast Breast CRL-2273 CHP-212 Neuroblast CNS CNS CRL-2062 DMS53 Small cell Lung SCLC ACC 231 EFM-19 Epithelioid Breast Breast ** ACC 317 KPL-1 N/A Breast Breast ** ACC 403 MT-3 Epithelial Breast Breast ** HTB-178 NC1-H596 Epithelial Lung Lung HTB-3 SCaBER Epithelial Bladder Bladder HTB-58 SKMES1 Squamous Cell Lung Lung HTB-13 SW1783 Fibroblast CNS CNS ACC 291 U-138MG Fibroblastoid CNS Glioblastoma ** CRL-2505 22Rv1 Epithelial Prostate Prostate ACC 143 BPH1 Epithelioid Prostate Prostate ** HTB-54 Calu1 Squamous Cell Lung Lung HTB-75 CaOV3 Epithelial Female GU Ovary CCL-138 Detroit 562 Epithelial Head & Neck H&N CRL-7920 DoTc24510 Epithelial Female GU Cervix HTB-81 DU145 Epithelial Prostate Prostate HTB-135 HS 746T Epithelial Colon/GI Stomach HTB-32 HT-3 Epithelial Female GU Cervix CCL-253 NCl-H508 Epithelial Colon/GI Colon CRL-1671 RL95-2 Epithelial Female GU Uterus CRL-1628 SCC-25 Epithelial Head & Neck H&N HTB-77 SKOV3 Epithelial Female GU Ovary CCL-238 SW1417 Epithelial Colon/GI Colon CCL-235 SW837 Epithelial Colon/GI Colon HTB-117 SW 954 Epithelial Female GU Vulva HTB-118 SW 962 Mixed Female GU Vulva - Cells were grown in RPMI1640, 10% FBS, 2 mM L-alanyl-L-Glutamine, 1 mM Na Pyruvate in a humidified atmosphere of 5% CO2 at 37° C. Cells were seeded into 384-well plates and incubated in a humidified atmosphere of 5% CO2 at 37° C. Compounds NE10214 and GPB-032 were added 24 hours post cell seeding. At the same time, a time zero untreated cell plate was generated.
- After a 72 hour incubation period, cells were fixed and stained with fluorescently labeled antibodies and nuclear dye to allow visualization of nuclei, apoptotic cells and mitotic cells. Apoptotic cells were detected using an anti-active caspase-3 antibody. Mitotic cells were detected using an anti phospho-histone-3 antibody.
- The concentrated Au suspension (NE10214, also “
Compound 1”) and the concentrated bi-metallic suspension AuPt (GPB-032, also “Compound 2”) were diluted as shown in Table 26 below and assayed over 9 concentrations from the highest test concentration to the lowest test concentration. When the two test compounds were added to the growth medium they became diluted by the growth media. The actual atomic concentrations of the metallic components (i.e., Au in NE10214; and Au+Pt in GPB-032) in the growth media are shown in Table 26 as “In Vitro Conc microM”. - Automated fluorescence microscopy was carried out using a GE Healthcare
IN Cell Analyzer 1000, and images were collected with a 4× objective. -
TABLE 26 Initial and In Vitro Concentrations Compound 1 (NE10214) Compound 2 (GPB-032) initial In Vitro initial In Vitro sample conc., Conc sample conc., Conc ID ppm microM ID ppm microM 1-1 981 701 2-1 982 701 1-2 800 571 2-2 800 571 1-3 600 429 2-3 600 429 1-4 400 286 2-4 400 286 1-5 200 143 2-5 385 275 1-6 80 57 2-6 180 129 1-7 40 29 2-7 40 29 1-8 20 14 2-8 20 14 1-9 8 5.7 2-9 8 5.7 1-10 vehicle vehicle 2-10 vehicle vehicle - Twelve bit tiff images were acquired using the
InCell Analyzer 1000 3.2 and analyzed with Developer Toolbox 1.6 software. EC50 and IC50 values were calculated using nonlinear regression to fit data to a sigmoidal 4 point, 4 parameter One-Site dose response model, where: y (fit)=A+[(B−A)/(1+((C/x){circumflex over ( )}D))]. Curve-fitting, EC50/IC50 calculations and report generation are performed using a custom data reduction engine MathIQ based software (AIM). -
TABLE 27 Summary table for vehicle background Relative cell count (POC) Apoptosis (fold induction) Mitosis (fold induction) Plate # Cell line Mean StdDev CV Mean StdDev CV Mean StdDev CV Doublings 4 HS 746T 100.00 3.40 0.03 1.00 0.21 0.21 1.00 0.28 0.28 2.17 4 NCI-H596 100.00 4.07 0.04 1.00 0.30 0.30 0.98 0.61 0.62 2.08 4 NCI-H508 100.00 3.20 0.03 1.00 0.26 0.26 1.00 0.15 0.15 2.92 4 HT-3 100.00 2.68 0.03 0.99 0.28 0.28 0.99 0.17 0.17 2.50 4 KPL-1 100.00 8.31 0.08 1.01 0.59 0.59 1.01 0.18 0.18 2.40 4 EFM-19 100.00 6.45 0.06 1.00 0.26 0.26 1.00 0.15 0.15 1.10 4 DU145 100.00 3.35 0.03 1.00 0.44 0.44 1.00 0.10 0.10 3.07 4 SKMES1 100.00 3.81 0.04 1.00 0.45 0.45 1.00 0.12 0.12 3.46 4 SKOV3 100.00 3.14 0.03 1.00 0.24 0.24 1.00 0.16 0.16 1.47 4 SW837 100.00 6.10 0.06 1.01 0.25 0.25 1.00 0.15 0.15 2.26 4 SCaBER 100.00 3.07 0.03 1.00 0.38 0.38 1.00 0.17 0.17 3.29 4 U-138MG 100.00 2.89 0.03 1.00 0.45 0.45 0.99 0.24 0.25 2.63 4 MT-3 100.00 6.96 0.07 1.00 0.29 0.29 1.00 0.12 0.12 3.16 4 RL95-2 100.00 4.68 0.05 1.00 0.30 0.30 1.00 0.13 0.13 1.76 4 SCC-25 100.00 5.11 0.05 1.01 0.36 0.36 1.00 0.14 0.14 3.08 4 SW962 100.00 5.43 0.05 1.01 0.32 0.32 1.00 0.29 0.29 1.99 4 SW954 100.00 6.77 0.07 1.00 0.26 0.26 1.00 0.15 0.15 2.37 4 647-V 100.00 5.46 0.05 1.00 0.30 0.30 1.00 0.12 0.12 4.05 4 BHT-101 100.00 6.02 0.06 0.99 0.32 0.32 1.00 0.13 0.13 3.89 4 BPH1 100.00 4.60 0.05 1.00 0.28 0.28 1.00 0.13 0.13 3.73 4 SW1783 100.00 4.26 0.04 1.00 0.30 0.30 1.00 0.26 0.26 1.55 4 SW1417 100.00 2.70 0.03 1.00 0.23 0.23 1.00 0.13 0.13 1.92 4 22Rv1 100.00 6.12 0.06 1.00 0.27 0.26 1.00 0.11 0.11 2.40 4 DoTc2 4510 100.00 7.65 0.08 1.01 0.28 0.28 1.00 0.12 0.12 2.21 4 DMS53 100.00 2.22 0.02 1.00 0.38 0.38 1.00 0.12 0.12 1.81 4 CaOV3 100.00 3.09 0.03 1.00 0.19 0.19 1.00 0.12 0.12 1.94 4 Detroit 562 100.00 9.02 0.09 1.01 0.22 0.22 1.01 0.15 0.15 3.13 4 BT474 100.00 1.41 0.01 1.00 0.34 0.34 1.00 0.23 0.23 1.36 4 Calu1 100.00 2.60 0.03 1.00 0.55 0.55 1.00 0.15 0.15 2.41 4 CHP-212 100.00 3.05 0.03 1.00 0.26 0.26 1.00 0.18 0.18 2.55 -
TABLE 28 Performance Summary for Compounds 1 (NE10214) and 2 (GPB-032) Relative cell Relative cell Apoptosis 5X G2/M cell G1/S cell Max G2/M count EC50 count IC50 Fold Induction Max Apoptosis cycle block cycle block cell cycle Plate # Compound Cell line (ppm) (ppm) (ppm) Fold Induction (ppm) (ppm) block 4 1 SW1417 >9.81E+02 >9.81E+02 N/A 1.20 N/A N/A 0.96 4 1 SW1783* 6.37E+02* 6.37E+02* N/A 0.82 N/A 6.65E+01* 0.80 4 1 22Rv1 >9.81E+02 >9.81E+02 N/A 1.33 N/A N/A 0.95 4 1 647-V >9.81E+02 >9.81E+02 N/A 2.10 N/A N/A 0.91 4 1 SW954* 2.44E+02* 2.94E+02* N/A 0.97 N/A 7.63E+01* 1.05 4 1 SW962 8.00E+02 8.00E+02 N/A 0.65 N/A N/A 1.40 4 1 BHT-101* 7.52E+02* 7.52E+02* N/A 2.75 N/A 7.67E+02* 0.98 4 1 BPH1 >9.81E+02 >9.81E+02 N/A 1.65 N/A N/A 0.95 4 1 BT474 >9.81E+02 >9.81E+02 N/A 2.48 N/A N/A 0.97 4 1 Calu1* 5.27E+02* 5.27E+02* N/A 2.53 N/A 1.05E+02* 0.83 4 1 CHP-212* 4.37E+02* 4.37E+02* N/A 1.02 N/A N/A 1.02 4 1 CaOV3 >9.81E+02 >9.81E+02 N/A 1.35 N/A N/A 1.45 4 1 DoTc2 4510 >9.81E+02 >9.81E+02 N/A 1.42 N/A N/A 0.88 4 1 DMS53 >9.81E+02 >9.81E+02 N/A 2.02 N/A N/A 0.86 4 1 Detroit 562* 2.30E+02* 8.65E+02 N/A 1.33 N/A 6.96E+02* 1.00 4 1 DU145 8.88E+02 8.88E+02 N/A 2.86 N/A N/A 0.92 4 1 EFM-19* 1.71E+02* 1.71E+02* N/A 1.90 N/A 5.56E+02* 1.22 4 1 SKMES1* 6.60E+02* 6.60E+02* N/A 1.63 N/A N/A 0.97 4 1 NCI-H508 >9.81E+02 >9.81E+02 N/A 1.06 N/A 9.21E+02 1.01 4 1 NCI-H596 >9.81E+02 >9.81E+02 N/A 1.08 N/A N/A 1.81 4 1 HS 746T* 5.02E+02* 5.02E+02* N/A 0.88 N/A 1.23E+02* 1.08 4 1 HT-3 >9.81E+02 >9.81E+02 N/A 0.80 N/A N/A 1.01 4 1 KPL-1 9.02E+02 9.02E+02 N/A 3.54 N/A 8.09E+02 1.31 4 1 MT-3 >9.81E+02 >9.81E+02 N/A 0.83 N/A N/A 1.03 4 1 RL95-2 >9.81E+02 >9.81E+02 N/A 1.48 N/A N/A 0.96 4 1 SCC-25* 4.60E+02* 4.60E+02* N/A 1.52 N/A 9.39E+01* 0.84 4 1 SCaBER* 6.20E+01* >9.81E+02 N/A 1.12 N/A N/A 0.85 4 1 SKOV3* >9.81E+02 >9.81E+02 N/A 0.83 N/A 2.66E+02* 1.20 4 1 SW837 >9.81E+02 >9.81E+02 N/A 1.01 N/A 8.14E+02 0.80 4 1 U-138MG* 6.35E+02* >9.81E+02 N/A 0.99 N/A 7.97E+01* 0.75 4 2 SW1417 9.54E+02 9.54E+02 N/A 1.39 N/A N/A 0.95 4 2 SW1783 >9.82E+02 >9.82E+02 N/A 1.06 N/A 5.91E+02* 0.92 4 2 22Rv1* 4.75E+02* 4.75E+02* 6.08E+02* 4.77* N/A 5.58E+02* 0.89 4 2 647-V >9.82E+02 >9.82E+02 N/A 4.89 N/A N/A 0.90 4 2 SW954* 5.22E+02* 5.22E+02* N/A 1.15 N/A N/A 0.87 4 2 SW962* 5.25E+02* 5.25E+02* 5.98E+02* 5.39* 5.81E+02* N/A 4.09* 4 2 BHT-101* 5.83E+02* 5.83E+02* 8.67E+02* 7.34* N/A N/A 1.00 4 2 BPH1* 5.80E+02* 5.80E+02* N/A 2.85 N/A 8.28E+02 0.92 4 2 BT474* 7.28E+02* 7.28E+02* 5.91E+02* 6.70* N/A N/A 1.01 4 2 Calu1* 4.36E+02* 4.36E+02* N/A 3.40 N/A N/A 0.87 4 2 CHP-212* 5.11E+02* 5.11E+02* N/A 1.60 N/A 6.77E+02* 0.88 4 2 CaOV3* 5.64E+02* 5.74E+02* 9.67E+02* 5.21* 5.90E+02* N/A 3.64* 4 2 DoTc2 4510* 4.54E+02* 4.54E+02* 5.89E+02* 5.59* N/A N/A 0.95 4 2 DMS53 >9.82E+02 >9.82E+02 N/A 2.86 N/A N/A 0.86 4 2 Detroit 562* 5.32E+02* 5.63E+02* N/A 2.71 N/A 5.50E+02* 0.97 4 2 DU145* 4.57E+02* 4.60E+02* 4.82E+02* 35.16* N/A N/A 1.07 4 2 EFM-19* 1.10E+02* 1.10E+02* N/A 3.83 5.60E+02* N/A 7.50* 4 2 SKMES1* 6.86E+02* 6.86E+02* N/A 1.68 N/A 8.77E+02* 0.97 4 2 NCI-H508* 8.79E+02* 8.79E+02* N/A 1.56 N/A 7.84E+02* 0.99 4 2 NCI-H596 >9.82E+02 >9.82E+02 N/A 1.50 N/A N/A 1.90 4 2 HS 746T* 4.25E+02* >9.82E+02 N/A 0.96 N/A N/A 1.02 4 2 HT-3* 5.71E+02* 5.71E+02* N/A 2.49 N/A 4.58E+02* 1.11 4 2 KPL-1* 9.00E+02 9.00E+02 3.51E+02* 14.20* N/A 9.21E+02* 1.30 4 2 MT-3 9.35E+02 9.35E+02 N/A 2.63 N/A N/A 1.07 4 2 RL95-2* 4.99E+02* 5.01E+02* N/A 2.96 5.28E+02* N/A 6.80* 4 2 SCC-25* 4.89E+02* 4.89E+02* N/A 1.28 N/A N/A 1.01 4 2 SCaBER* 7.40E+02* 7.40E+02* N/A 1.29 N/A 6.52E+02* 0.91 4 2 SKOV3 >9.82E+02 >9.82E+02 N/A 2.28 N/A N/A 0.94 4 2 SW837* 5.69E+02* 5.69E+02* N/A 1.00 7.43E+02* N/A 2.22* 4 2 U-138MG* >9.82E+02 >9.82E+02 N/A 1.11 N/A 5.29E+02* 0.88 An “*” in column 3 “Cell Line” indicates significant anti-cancer activity in that tumor cell line. An “*” in columns An “*” in columns An “*” in column - The multiplexed cytotoxicity assay used a cell image based analysis technique where cells were fixed and stained with fluorescently labeled antibodies and nuclear dye as mentioned above.
- Cell proliferation was measured by the signal intensity of the incorporated nuclear dye. The cell proliferation assay output is referred to as the relative cell count. To determine the cell proliferation end point, the cell proliferation data output was transformed to percent of control (POC) using the following formula:
-
POC=relative cell count (compound wells)/relative cell count (vehicle wells)×100 - Relative cell count IC50 is the test compound concentration at 50% of maximal possible response. A relative cell count EC50 is the test compound concentration at the curve inflection point or half the effective response (parameter C of the fitted curve solution). GI50 is the concentration needed to reduce the observed growth by half. This is the concentration that inhibits the growth midway between untreated cells and the number of cells seeded in the well (Time zero value).
- Time zero non-treated plate is used to determine number of doublings in 72 hour assay period: Number of doublings in 72 hours=LN[Cell number (72 hrs end point) *Cell number (time zero)]/LN(2)
- The output of each biomarker is fold increase over vehicle background normalized to the relative cell count in each well.
- The activated caspase-3 marker labels cells from early to late stage apoptosis. The output is shown as a fold increase of apoptotic cells over vehicle background normalized to the relative cell count in each well. Concentrations of test compound that cause a 5-fold induction in the caspase-3 signal indicates significant apoptosis induction. Wells with concentrations higher than the relative cell count IC95 are eliminated from the caspase3 induction analysis.
- The phospho-histone-3 marker labels mitotic cells. The output is shown as a fold induction of mitotic cells over vehicle background normalized to the relative cell count in each well. When the fold induction of mitotic cell signal over background is ˜1, there is “no effect” on the cell cycle. Two or more fold increase in phospho-histone-3 signal over vehicle background indicates significant test compound induction of mitotic block.
- Two or more fold decrease in the phospho-histone-3 signal may indicate G1/S block only when cytotoxicity levels are below the measured relative cell count IC95. When 2 or more fold decrease in the phospho-histone-3 signal are observed at concentrations higher than the relative cell count IC95, the decrease in mitotic cell counts are most likely due to a more general cytotoxicity effect rather than a true G1/S phase block. Wells with concentrations higher than the relative cell count IC95 are eliminated from the phospho-histone-3 analysis.
-
-
- Cell proliferation measured by relative cell counts
- Apoptosis:
- >5-fold increase in activated caspase-3 signal indicates an apoptotic response
- Mitosis:
- >2-fold increase in phospho-histone-3 indicates mitotic block
- <2-fold decrease in phospho-histone-3 indicates G1/S block
- Because the compounds are at relatively low concentration levels in vitro, most concentrations provided were too low to obtain IC50 results. As concentration levels increase, activity becomes clearly apparent with both compounds in many of the tumor cell lines tested. Table 28 entitled, “Perfomance Summary for Compounds 1 (NE10214) and 2 (GPB-032)” above highlights in Column 3 (“Cell Line”) a “*” for each tumor cell line where significant anti-cancer activity was demonstrated for each compound/cell line combination.
- The data summarized in Table 28 clearly demonstrate significant anti-cancer activity in response to treatment with the concentrated Au suspension (NE10214) in 13 of 30 tumor cell lines tested, and in 23 of of the 30 tumor cell lines treated with the concentrated Au—Pt bi-metallic suspension (GPB-032).
- Equally important, the concentrated Au suspension and the concentrated Au—Pt bi-metallic suspension show distinctly different patterns of the presence of anti-cancer activity, and distinctly different patterns of the type of anti-cancer activity, across the thirty different tumor cell lines.
- Reference is now made to
FIGS. 32 a -32 ad. These figures show graphically the difference in performance ofcompound 1 andcompound 2 against each of the 30 cell lines tested. Specifically, comparisons are set forth for each of “Relative Cell Count %”, “Apoptosis (fold induction)” and “Mitosis (fold induction)”. The data show that there is a significant elevation in apoptosis induction in eight different tumor cell lines treated with the concentrated Au—Pt bi-metallic suspension (GPB-032), but this kind of activity is not shown in any of the tumor cell lines treated with the concentrated Au compound (NE10214). - Significant Elevation of Apoptosis Induction is clearly present in the eight tumor cell lines set forth below treated with the concentrated Au—Pt bi-metallic suspension, but in none with the concentrated Au suspension:
-
22Rv1 Prostate SW962 Vulva BHT 101 Endocrine BT474 Breast CaOV- 3 Ovary DoTc2 4510 Cervix Du 145 Prostate KPL-1 Breast. - Secondly, there is significant induction of Mitosis block in the five different tumor cell lines treated with the concentrated Au—Pt bi-metallic suspension (GPB-032), but this kind of activity is not shown in any of the cell lines when treated with the concentrated Au suspension (NE10214).
- Significant Induction of Mitotic Block is present in five types of tumor cell lines set forth below treated with the concentrated Au—Pt bi-metallic suspension, but in none treated with the concentrated Au suspension:
-
SW837 Rectum RL95-2 Uterus EFM-19 Breast SW962 Vulva CAOV3 Ovary - Third, the concentrated Au—Pt bi-metallic suspension shows significant anti-cancer activity in twelve tumor cell lines where the concentrated Au compound showed no activity at all, and the concentrated Au suspension is effective in two additional tumor cell lines where the concentrated AuPt bi-metallic suspension shows no activity at all,—so in fourteen of thirty tumor cell lines, there is no shown overlap in the presence of any kind of anti-cancer activity.
- Furthermore, in the twenty-five of thirty cell lines where either the concentrated Au suspension or the concentrated Au—Pt bi-metallic suspension, or both, showed anti-cancer activity, in only four (4/30=13%) do both compounds have the same pattern or type of anti-cancer activity. In twenty-three of twenty-seven cases, the pattern of activity is distinctly different.
- In summary,
1) Significant Level of Anti-Cancer Activity: either the concentrated Au suspension, or the concentrated AuPt bi-metallic suspension, or both compounds, had significant anti-cancer activity against twenty-five of the thirty (25/30=83%) tumor cell lines tested;
2) Distinctly Different Patterns of Anti-Cancer Activity: the pattern of anti-cancer activity of the two compounds (Au and AuPt) was distinctly different in twenty-one of the twenty-five tumor cell lines where there wasactivity 21/25→84% had distinctly different patterns of activity as between the concentrated Au suspension and the concentrated Au—Pt bi-metallic suspension. - This Example demonstrates the efficacy of several orally administered inventive compositions in a mouse xenograft cancer model. Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein. The Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm3 in size. The Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle. Once the tumors in the recipient mice had reached a measurable size of about 4×4 mm, as measured by calipers placed against each mouse skin, the recipient mice were randomly placed into treatment groups, 3 per group and the oral treatment was started. Treatment was given exclusively via the drinking bottle shared between 3 mice in each group. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated at
day 24. The results of the Example are summarized inFIGS. 33 a -33 b. - Certain comparative nanocrystal suspensions and ionic solutions were prepared to compare to the bi-metallic Au—Pt nanocrystal suspensions.
- Briefly, GB-218 was prepared similarly to Example 1 resulting in a gold concentration of 7.6 ppm as measured by AAS. Additionally said solution was determined to have a hydrodynamic radius of 15.1 nm as measured by the Viscotek. GB-219 was prepared similarly in regards to Example 1 wherein potassium hydroxide was replaced as the process enhancer for sodium bicarbonate at a concentration of 0.63 g/gallon (i.e., about 0.17 mg/mL). GB-219 had a gold concentration of 8.7 ppm as measured by AAS. Additionally said solution was determined to have a hydrodynamic radius of 18.3 nm as measured by the Viscotek.
- In addition, PB-39 was prepared similarly to Example 13 PB57001 example, resulting in a suspension of nanocyrystal platinum particles having a Pt concentration of 7.4 ppm. PB-22-C4 was prepared similarly to Example 13, wherein the applied frequency of 501AC was set to 80 Hz instead of 5 Hz to produce a solution comprising predominantly of Pt ionic species with a small amount of Pt nanocrystalline species. The concentration of sodium bicarbonate was 2.5 g/gallon (i.e., about 0.66 mg/mL). PB-22-C4 was then subsequently concentrated using an electrical hot plate to produce a Pt concentration of about 8.3 ppm.
-
- Species: Mice
- Strain: Balb/C immunodeficient mice
- Source: Harlan
- Gender and number: Female, 24
- Age: About 6-8 weeks old at the start of the study.
- Identification: Each mouse was given a unique identity number.
- Animal husbandry: On receipt, all animals were examined for external signs of ill-health and all unhealthy animals were excluded from further evaluation. Animals were housed in groups of three under specific pathogen free (spf) conditions, in a thermostatically monitored room (22±4° C.) in an animal unit. Animals were equilibrated under standard animal house conditions for at least 72 hours prior to use. The health status of the animals was monitored throughout this period and the suitability of each animal for experimental use was assessed prior to study start.
- Housing Animals were housed in groups of 3 per cage in a controlled room, to ensure correct temperature, humidity and 12 hour light/dark cycle for the duration of the study.
- Diet: Irradiated pellet diet and water was available ad libitum throughout the holding, acclimatisation and post-dose periods.
- HCT 116 cell line (ATCC CCL-247).
Phosphate buffered saline (“PBS”).
Test compounds: platinum nanocrystal suspension, gold nanocrystal suspension and Au—Pt bi-metallic suspension.
Positive control compound: cisplatin.
Negative control compound: drinking water. - Negative Control Group 1: Days 0-24, given normal drinking water.
Positive Control Group 2: Days 0-24, given normal drinking water; and given a daily cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”).
Treatment Group 3-6: Days 0-24, given test compounds as their drinking water. - a.) Preparation of Tumor Cells
- 1. Cells were grown in complete medium and all contaminants were excluded.
2. When the cells were approximately 70-80% confluent, then approximately 3-4 hours before harvesting, the old cell growth medium was replaced with fresh cell growth medium to remove any dead and/or detached cells.
3. The cell growth medium was once again removed and the cells were washed with PBS. A small amount (e.g., 10 ml) of trypsin-EDTA was then added. The cells were then dispersed in complete cell growth medium in a ratio of between 10/1 and 5/1. The dispersed cells and medium were thereafter immediately centrifuged at about 1500 rpm for about 5 minutes and were further washed twice with PBS and the cells were stored on ice.
4. The cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer.
5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 μL contained about 3×106 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site. - 1. Simultaneous with preparation of tumor growth cells, Balb/C mice had previously arrived and their health was checked.
2. All animals were allowed to acclimate for at least 72 hours.
3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation.
4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe.
5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm3.
6. The mice were then anesthetized and the tumors were harvested by using a scalpel and appropriately stored prior to being injected into the recipient mice.
Protocol B: Insertion of Tumors from Donor Mice into Recipient Mice
1. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag.
2. The recipient mice were allowed to acclimate for at least 72 hours.
3. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm3 in size. The 2 mm3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank). The tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm3 before treatment started atday 0. Treatments continued for 24 days or until the mouse was removed from the study and euthanized or the mouse died.
4. The tumor sizes and weights of the animals were determined daily until the end of the study atday 24. -
FIGS. 33 a and 33 b show graphically the results of the oral test.FIG. 33 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better. Further,FIG. 33 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better. - Table 29 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia. The Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
-
TABLE 29 Oral Treatment No. of Mice No. of Days Sample ID Removed into Study GB-218 1 9 1 14 1 18 PB-39 2 16 1 24 PB-22- C4 1 16 2 23 AuPt110810 1 23 2 24 GB-219 1 18 2 24 PtAu-111710-9 1 7 1 10 1 24 Cisplatin 3 24 Controls 1 15 1 22 1 24 - This Example demonstrates the efficacy of several intratumorally (“IT”) administered inventive metallic nanocrystal suspensions in a mouse xenograft cancer model. Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein. The Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm3 in size. The Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle. Once the tumors in the recipient mice had reached a measureable size of about 7×7 mm, as measured by calipers placed against each mouse skin, the recipient mice were randomly placed into treatment groups, 3 per group and the “IT” treatment was started. Treatment was given exclusively by needle injection into the tumor twice a day. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated at
day 30. The results of the Example are summarized inFIG. 34 a -34 b. - Certain comparative nanocrystal suspensions and ionic solutions were prepared to compare to the bi-metallic Au—Pt nanocrystal suspensions.
- Briefly, GB-218 was prepared similarly to Example 1 resulting in a gold concentration of 7.6 ppm as measured by AAS. Additionally said solution was determined to have a hydrodynamic radius of 15.1 nm as measured by the Viscotek. GB-219 was prepared similarly in regards to Example 1 wherein potassium hydroxide was replaced as the process enhancer for sodium bicarbonate at a concentration of 0.63 g/gallon (i.e., about 0.17 mg/mL). GB-219 had a gold concentration of 8.7 ppm as measured by AAS. Additionally said solution was determined to have a hydrodynamic radius of 18.3 nm as measured by the Viscotek.
- In addition, PB-39 was prepared similarly to Example 13 PB57001 example, resulting in a suspension of nanocyrystal platinum particles having a Pt concentration of 7.4 ppm. PB-22-C4 was prepared similarly to Example 13, wherein the applied frequency of 501AC was set to 80 Hz instead of 5 Hz to produce a solution comprising predominantly of Pt ionic species with a small amount of Pt nanocrystalline species. The concentration of sodium bicarbonate was 2.5 g/gallon (i.e., about 0.66 mg/mL). PB-22-C4 was then subsequently concentrated using an electrical hot plate to produce a Pt concentration of about 8.3 ppm.
-
- Species: Mice
- Strain: Balb/C immunodeficient mice
- Source: Harlan
- Gender and number: Female, 24
- Age: About 6-8 weeks old at the start of the study.
- Identification: Each mouse was given a unique identity number.
- Animal husbandry: On receipt, all animals were examined for external signs of ill-health and all unhealthy animals were excluded from further evaluation. Animals were housed in groups of three under specific pathogen free (spf) conditions, in a thermostatically monitored room (22±4° C.) in an animal unit. Animals were equilibrated under standard animal house conditions for at least 72 hours prior to use. The health status of the animals was monitored throughout this period and the suitability of each animal for experimental use was assessed prior to study start.
- Housing Animals were housed in groups of 3 per cage in a controlled room, to ensure correct temperature, humidity and 12 hour light/dark cycle for the duration of the study.
- Diet: Irradiated pellet diet and water was available ad libitum throughout the holding, acclimatization and post-dose periods.
- HCT 116 cell line (ATCC CCL-247).
Phosphate buffered saline (“PBS”).
Test compounds: platinum nanocrystal suspension, gold nanocrystal suspension and Au—Pt bi-metallic suspension.
Positive control compound: cisplatin.
Negative control compound: drinking water. - Negative Control Group 1: Days 0-30, saline injection twice a day, with a total of 100 μl in each tumor divided between 2-3 injection points; (given normal drinking water to drink).
Positive Control Group 2: Days 0-30,cisplatin injection 8 mg/kg given once a day into the peritoneum (IP) (given normal drinking water to drink).
Treatment Group 3-6: Days 0-30, nanocrystal formulation injection twice a day, with a total of 100 μl in each tumor divided between 2-3 injection points; (given normal drinking water to drink). - a.) Preparation of Tumor Cells
- 1. Cells were grown in complete medium and all contaminants were excluded.
2. When the cells were approximately 70-80% confluent, then approximately 3-4 hours before harvesting, the old cell growth medium was replaced with fresh cell growth medium to remove any dead and/or detached cells.
3. The cell growth medium was once again removed and the cells were washed with PBS. A small amount (e.g., 10 ml) of trypsin-EDTA was then added. The cells were then dispersed in complete cell growth medium in a ratio of between 10/1 and 5/1. The dispersed cells and medium were thereafter immediately centrifuged at about 1500 rpm for about 5 minutes and were further washed twice with PBS and the cells were stored on ice.
4. The cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer.
5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 μL contained about 3×106 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site. - 1. Simultaneous with preparation of tumor growth cells, Balb/C mice had previously arrived and their health was checked.
2. All animals were allowed to acclimate for at least 72 hours.
3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation.
4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe.
5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm3.
6. The mice were then anesthetized and the tumors were harvested by using a scalpel and appropriately stored prior to being injected into the recipient mice.
Protocol B: Insertion of Tumors from Donor Mice into Recipient Mice
5. Additional Blab/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag.
6. The mice were allowed to acclimate for at least 72 hours.
7. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm3 in size. The 2 mm3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank). The tumors were permitted to grow in the recipient mice until they reached a size of about 7×7 mm before treatment started atday 0. Treatments continued for 30 days or until the mouse was removed from the study and euthanized or the mouse died.
8. The tumor sizes and weights of the animals were determined daily until the end of the study atday 24.
Protocol C: Intertumoral Injection into Recipient Mice
1. Each tumor in each recipient mouse was injected twice daily (about 12 hours apart) with about 100 μl of either negative control, positive control or test compound. The needle used for injection was either a 25 Ga or 26 Ga needle. Depending on the tumor size, there were either 2 or 3 injection points for each tumor. -
FIGS. 34 a and 34 b shows graphically the results of the IT test.FIG. 34 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better. Further,FIG. 34 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better. - Table 30 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia. The Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
-
TABLE 30 IT Treatment No. of Mice No. of Days Sample ID Removed into Study GB-218 1 9 1 11 1 15 PB-39 1 7 1 15 1 28 PB-22- C4 2 11 1 30 AuPt110810 2 15 1 23 GB-219 1 14 1 17 1 25 PtAu-111710-9 2 14 1 30 Cisplatin 1 15 1 18 1 30 Controls 1 15 1 16 - This Example demonstrates the relative efficacy of four orally administered inventive metallic nanocrystal suspensions in a mouse xenograft cancer model. Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein. The Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm3 in size. The Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle. Once the tumors in the recipient mice had reached a measurable size of about 4×4 mm, as measured by calipers placed against each mouse skin, the recipient mice were randomly placed into treatment groups, 6 per group and the oral treatment was started. 6 mice were in the positive control group (“Cisplatin”) and 6 mice were in the negative control group and received only water (“Control”). Treatment was given exclusively via the drinking bottle shared between the mice in each Treatment group. Cisplatin was given by intraperitoneal injection on
day 0. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated as scheduled. The results of the Example are summarized inFIGS. 35 a-35 b . -
- Species: Mice
- Strain: Balb/C immunodeficient mice
- Source: Harlan
- Gender and number: Female, 36
- Age: About 6-8 weeks old at the start of the study.
- Identification: Each mouse was given a unique identity number.
- Animal husbandry: On receipt, all animals were examined for external signs of ill-health and all unhealthy animals were excluded from further evaluation. Animals were housed in groups of three under specific pathogen free (spf) conditions, in a thermostatically monitored room (22±4° C.) in an animal unit. Animals were equilibrated under standard animal house conditions for at least 72 hours prior to use. The health status of the animals was monitored throughout this period and the suitability of each animal for experimental use was assessed prior to study start.
- Housing Animals were housed in groups of 3 per cage in a controlled room, to ensure correct temperature, humidity and 12 hour light/dark cycle for the duration of the study.
- Diet: Irradiated pellet diet and water was available ad libitum throughout the holding, acclimatisation and post-dose periods.
- HCT 116 cell line (ATCC CCL-247).
Phosphate buffered saline (“PBS”).
Test compounds: platinum nanocrystal suspension, gold nanocrystal suspension and Au—Pt bi-metallic suspension.
Positive control compound: cisplatin.
Negative control compound: drinking water. - Negative Control Group 1: Days 0-24, given normal drinking water.
Positive Control Group 2: Days 0-24, given normal drinking water; and given a one-time cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”) onday 0.
Treatment Group 3-6: Days 0-24, given test compounds as their drinking water. - a.) Preparation of Tumor Cells
- 1. Cells were grown in complete medium and all contaminants were excluded.
2. When the cells were approximately 70-80% confluent, then approximately 3-4 hours before harvesting, the old cell growth medium was replaced with fresh cell growth medium to remove any dead and/or detached cells.
3. The cell growth medium was once again removed and the cells were washed with PBS. A small amount (e.g., 10 ml) of trypsin-EDTA was then added. The cells were then dispersed in complete cell growth medium in a ratio of between 10/1 and 5/1. The dispersed cells and medium were thereafter immediately centrifuged at about 1500 rpm for about 5 minutes and were further washed twice with PBS and the cells were stored on ice.
4. The cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer.
5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 μL contained about 3×106 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site. - b.) Injection and Growth of Tumor Cells
- 1. Simultaneous with preparation of tumor growth cells, Balb/C mice had previously arrived and their health was checked.
2. All animals were allowed to acclimate for at least 72 hours.
3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation.
4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe.
5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm3.
6. The mice were then anesthetized and the tumors were harvested by using a scalpel and appropriately stored prior to being injected into the recipient mice.
Protocol B: Insertion of Tumors from Donor Mice into Recipient Mice
9. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag.
10. The recipient mice were allowed to acclimate for at least 72 hours.
11. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm3 in size. The 2 mm3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank). The tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm3 before treatment started atday 0. Treatments continued for 24 days or until the mouse was removed from the study and euthanized or the mouse died.
12. The tumor sizes and weights of the animals were determined daily until the end of the study atday 24. -
FIGS. 35 a and 35 b show graphically the results of the oral test.FIG. 35 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better. Further,FIG. 35 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better. - Table 31 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia. The Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
-
TABLE 31 Oral Treatment No. of Mice No. of Days Sample ID Removed into Study PGT001 1 11 PGB002 1 14 1 18 1 19 1 22 PB56001 1 14 1 15 1 18 1 19 1 20 1 21 PB57001 4 11 Cisplatin 1 11 1 13 1 14 2 18 1 22 Control 4 12 1 15 1 18 1 19 - Table 32 provides a comparison of the doubling time (RTV2) for each group in the study. In addition, table 32 also lists the growth delay in days, maximum percent weight loss and statistical significance of the data.
-
TABLE 32 Mean Median Time Time Growth Group to RTV2 to RTV2 Delay Maximum % Number (days) (days) (days) Significance Weight Loss 1 3.9 3.6 — — 1 (d4) 2 6.7 5.2 1.6 p < 0.05 4 (d5) 3 8.3 7.6 4.0 p < 0.01 2 (d8) 4 5.7 5.6 2.0 p < 0.05 2 (d11) 5 5.0 4.4 0.8 p > 0.05 ns 3 (d6) 6 5.9 5.5 1.9 p > 0.05 ns 4 (d8) - This Example demonstrates the relative efficacy of three orally administered inventive metallic nanocrystal suspensions in a mouse xenograft cancer model relative to Cisplatin. Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein. The Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm3 in size. The Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle. Once the tumors in the recipient mice had reached a measurable size of about 4×4 mm, as measured by calipers placed against each mouse skin, the recipient mice were randomly placed into treatment groups, 8 per group and the oral treatment was started. 8 mice were in the positive control group (“Cisplatin”) and 8 mice were in the negative control group and received only water (“Control”). Treatment was given exclusively via the drinking bottle shared between the mice in each Treatment group. Cisplatin was given by intraperitoneal injection on
day 0. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated as scheduled. The results of the Example are summarized inFIGS. 36 a-36 b . -
- Species: Mice
- Strain: Balb/C immunodeficient mice
- Source: Harlan
- Gender and number: Female, 36
- Age: About 6-8 weeks old at the start of the study.
- Identification: Each mouse was given a unique identity number.
- Animal husbandry: On receipt, all animals were examined for external signs of ill-health and all unhealthy animals were excluded from further evaluation. Animals were housed in groups of three under specific pathogen free (spf) conditions, in a thermostatically monitored room (22±4° C.) in an animal unit. Animals were equilibrated under standard animal house conditions for at least 72 hours prior to use. The health status of the animals was monitored throughout this period and the suitability of each animal for experimental use was assessed prior to study start.
- Housing Animals were housed in groups of 3 per cage in a controlled room, to ensure correct temperature, humidity and 12 hour light/dark cycle for the duration of the study.
- Diet: Irradiated pellet diet and water was available ad libitum throughout the holding, acclimatisation and post-dose periods.
- HCT 116 cell line (ATCC CCL-247).
Phosphate buffered saline (“PBS”).
Test compounds: Au—Pt bi-metallic nanocrystal suspensions.
Positive control compound: cisplatin.
Negative control compound: drinking water. - Negative Control Group 1: Days 0-21, given normal drinking water.
Positive Control Group 2: Days 0-21, given normal drinking water; and given a one-time cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”) onday 0.
Treatment Group 3-5: Days 0-21, given test compounds as their drinking water. - a.) Preparation of Tumor Cells
- 1. Cells were grown in complete medium and all contaminants were excluded.
2. When the cells were approximately 70-80% confluent, then approximately 3-4 hours before harvesting, the old cell growth medium was replaced with fresh cell growth medium to remove any dead and/or detached cells.
3. The cell growth medium was once again removed and the cells were washed with PBS. A small amount (e.g., 10 ml) of trypsin-EDTA was then added. The cells were then dispersed in complete cell growth medium in a ratio of between 10/1 and 5/1. The dispersed cells and medium were thereafter immediately centrifuged at about 1500 rpm for about 5 minutes and were further washed twice with PBS and the cells were stored on ice.
4. The cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer.
5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 μL contained about 3×106 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site. - b.) Injection and Growth of Tumor Cells
- 1. Simultaneous with preparation of tumor growth cells, Balb/C mice had previously arrived and their health was checked.
2. All animals were allowed to acclimate for at least 72 hours.
3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation.
4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe.
5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm3.
6. The mice were then anesthetized and the tumors were harvested by using a scalpel and appropriately stored prior to being injected into the recipient mice.
Protocol B: Insertion of Tumors from Donor Mice into Recipient Mice
13. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag.
14. The recipient mice were allowed to acclimate for at least 72 hours.
15. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm3 in size. The 2 mm3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank). The tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm3 before treatment started atday 0. Treatments continued for
21 days or until the mouse was removed from the study and euthanized or the mouse died.
16. The tumor sizes and weights of the animals were determined daily until the end of the study atday 21. -
FIGS. 36 a and 36 b show graphically the results of the oral test.FIG. 36 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better. Further,FIG. 36 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better. - Table 33 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia. The Sample IDs relate to compounds manufactured according to procedures discussed earlier herein.
-
TABLE 33 Oral Treatment No. of Mice No. of Days Group Number Sample ID Removed into Study 3 PGT024 1 15 3 16 1 17 1 21 4 PGT025 1 4 1 14 2 15 2 16 5 PGT026 1 11 1 14 1 15 2 21 2 Cisplatin 1 9 1 15 1 Control 1 15 4 16
Table 34 provides a comparison of the doubling time (RTV2) for each group in the study. In addition, table 34 also lists the growth delay in days, maximum percent weight loss and statistical significance of the data. -
TABLE 34 Median Mean Time Time Growth Group to RTV2 to RTV2 Delay Maximum % Number (days) (days) (days) Significance Weight Loss 1 3.3 3.5 — — 0 2 5.2 5.2 1.7 p < 0.05 5 (d7) 3 4.6 3.8 0.3 p < 0.05 ns 0 4 3.8 3.6 0.1 p < 0.05 ns 0 5 4.0 3.7 0.2 p > 0.05 ns 0 - This Example demonstrates the relative efficacy of three orally administered inventive Au—Pt bi-metallic nanoparticle suspensions in a mouse xenograft cancer model relative to Cisplatin. Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein. The Balb/C donor mice were used to grow H460 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm3 in size. The Balb/C recipient mice were given brief general anesthesia and then one H4602 mm3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle. Once the tumors in the recipient mice had reached a measurable size of about 4×4 mm, as measured by calipers placed against each mouse skin, the recipient mice were randomly placed into treatment groups, 8 per group and the oral treatment was started. 8 mice were in the positive control group (“Cisplatin”) and 8 mice were in the negative control group and received only water (“Control”). Treatment was given exclusively via the drinking bottle shared between the mice in each Treatment group. Cisplatin was given by intraperitoneal injection on
day 0. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated as scheduled. The results of the Example are summarized inFIGS. 37 a-37 b . -
- Species: Mice
- Strain: Balb/C immunodeficient mice
- Source: Harlan
- Gender and number: Female, 36
- Age: About 6-8 weeks old at the start of the study.
- Identification: Each mouse was given a unique identity number.
- Animal husbandry: On receipt, all animals were examined for external signs of ill-health and all unhealthy animals were excluded from further evaluation. Animals were housed in groups of three under specific pathogen free (spf) conditions, in a thermostatically monitored room (22±4° C.) in an animal unit. Animals were equilibrated under standard animal house conditions for at least 72 hours prior to use. The health status of the animals was monitored throughout this period and the suitability of each animal for experimental use was assessed prior to study start.
- Housing Animals were housed in groups of 3 per cage in a controlled room, to ensure correct temperature, humidity and 12 hour light/dark cycle for the duration of the study.
- Diet: Irradiated pellet diet and water was available ad libitum throughout the holding, acclimatisation and post-dose periods.
- H460 cell line (ATCC HTB-177).
Phosphate buffered saline (“PBS”).
Test compounds: Au—Pt bi-metallic nanocrystal suspensions.
Positive control compound: cisplatin.
Negative control compound: drinking water. - Negative Control Group 1: Days 0-21, given normal drinking water.
Positive Control Group 2: Days 0-21, given normal drinking water; and given a one-time cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”) onday 0.
Treatment Group 3-5: Days 0-21, given test compounds as their drinking water. - a.) Preparation of Tumor Cells
- 1. Cells were grown in complete medium and all contaminants were excluded.
2. When the cells were approximately 70-80% confluent, then approximately 3-4 hours before harvesting, the old cell growth medium was replaced with fresh cell growth medium to remove any dead and/or detached cells.
3. The cell growth medium was once again removed and the cells were washed with PBS. A small amount (e.g., 10 ml) of trypsin-EDTA was then added. The cells were then dispersed in complete cell growth medium in a ratio of between 10/1 and 5/1. The dispersed cells and medium were thereafter immediately centrifuged at about 1500 rpm for about 5 minutes and were further washed twice with PBS and the cells were stored on ice.
4. The cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer.
5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 μL contained about 3×106 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site. - b.) Injection and Growth of Tumor Cells
- 1. Simultaneous with preparation of tumor growth cells, Balb/C mice had previously arrived and their health was checked.
2. All animals were allowed to acclimate for at least 72 hours.
3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation.
4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26 gauge needle was subsequently added to the syringe.
5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm3.
6. The mice were then anesthetized and the tumors were harvested by using a scalpel and appropriately stored prior to being injected into the recipient mice.
Protocol B: Insertion of Tumors from Donor Mice into Recipient Mice
17. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag.
18. The recipient mice were allowed to acclimate for at least 72 hours.
19. H460 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm3 in size. The 2 mm3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank). The tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm3 before treatment started atday 0. Treatments continued for 24 days or until the mouse was removed from the study and euthanized or the mouse died.
20. The tumor sizes and weights of the animals were determined daily until the end of the study atday 21. -
FIGS. 37 a and 37 b show graphically the results of the oral test.FIG. 37 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better. Further,FIG. 37 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better. - Table 35 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia. The Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
-
TABLE 35 Oral Treatment No. of Mice No. of Days Group Number Sample ID Removed into Study 3 PGT024 1 14 2 15 1 16 1 18 4 PGT025 1 3 1 11 2 14 2 15 5 PGT026 2 11 1 14 1 18 2 Cisplatin 1 8 1 14 1 18 1 Control 1 14 4 15 3 18
Table 36 provides a comparison of the doubling time (RTV2) for each group in the study. In addition, table 34 also lists the growth delay in days, maximum percent weight loss and statistical significance of the data. -
TABLE 36 Median Mean Time Time Growth Group to RTV2 to RTV2 Delay Maximum % Number (days) (days) (days) Significance Weight Loss 1 2.3 2.5 — — 0 2 5.0 5.0 2.5 p < 0.01 6 (d3) 3 3.5 3.4 0.9 P < 0.05 0 4 3.5 3.0 0.5 p > 0.05 ns 0 5 3.7 3.6 1.1 P < 0.01 0 - This Example demonstrates the relative efficacy of one orally administered inventive Au—Pt bi-metallic nanocrystalline suspension in a mouse xenograft cancer model. Female Balb/C, immunologically deficient recipient mice (6-8 weeks old) had tumors implanted therein. The Balb/C donor mice were used to grow HCT116 tumors, which tumors were excised therefrom and subsequently sectioned into small fragments about 2 mm3 in size. The Balb/C recipient mice were given brief general anesthesia and then one HCT116 2 mm3 tumor fragment from the donor mice was implanted into each of the left and right flank of the recipient mice using a trocar needle. Once the tumors in the recipient mice had reached a measurable size of about 4×4 mm, as measured by calipers placed against each mouse skin, the recipient mice were randomly placed into treatment groups, 8 per group and the oral treatment was started. 8 mice were in the positive control group (“Cisplatin”) and 8 mice were in the negative control group and received only water (“Control”). Treatment was given exclusively via the drinking bottle shared between the mice in each Treatment group. Cisplatin was given by intraperitoneal injection on
day 0. Tumor size was assessed five times per week using a pair of calipers and mouse weight was also obtained by a scale, such measuring occurring until the mouse died (or was removed from the study) or the study was terminated as scheduled. The results of the Example are summarized inFIGS. 38 a-38 b -
- Species: Mice
- Strain: Balb/C immunodeficient mice
- Source: Harlan
- Gender and number: Female, 36
- Age: About 6-8 weeks old at the start of the study.
- Identification: Each mouse was given a unique identity number.
- Animal husbandry: On receipt, all animals were examined for external signs of ill-health and all unhealthy animals were excluded from further evaluation. Animals were housed in groups of three under specific pathogen free (spf) conditions, in a thermostatically monitored room (22±4° C.) in an animal unit. Animals were equilibrated under standard animal house conditions for at least 72 hours prior to use. The health status of the animals was monitored throughout this period and the suitability of each animal for experimental use was assessed prior to study start.
- Housing Animals were housed in groups of 3 per cage in a controlled room, to ensure correct temperature, humidity and 12 hour light/dark cycle for the duration of the study.
- Diet: Irradiated pellet diet and water was available ad libitum throughout the holding, acclimatisation and post-dose periods.
- HCT 116 cell line (ATCC CCL-247).
Phosphate buffered saline (“PBS”).
Test compounds: gold nanocrystal suspension NE-28-10X (NE-28 produced equivalent to - Positive control compound: cisplatin.
Negative control compound: drinking water. - Negative Control Group 1: Days 0-21, given normal drinking water.
Positive Control Group 2: Days 0-21, given normal drinking water; and given a one-time cisplatin dose of 8 mg/kg by intraperitoneal injection (“IP”) onday 0.
Treatment Group 3: Days 0-21, given test compounds as their drinking water. - a.) Preparation of Tumor Cells
- 1. Cells were grown in complete medium and all contaminants were excluded.
2. When the cells were approximately 70-80% confluent, then approximately 3-4 hours before harvesting, the old cell growth medium was replaced with fresh cell growth medium to remove any dead and/or detached cells.
3. The cell growth medium was once again removed and the cells were washed with PBS. A small amount (e.g., 10 ml) of trypsin-EDTA was then added. The cells were then dispersed in complete cell growth medium in a ratio of between 10/1 and 5/1. The dispersed cells and medium were thereafter immediately centrifuged at about 1500 rpm for about 5 minutes and were further washed twice with PBS and the cells were stored on ice.
4. The cells were then placed on a glass slide in the traditional manner and were counted using a hemocytometer.
5. Trypan-blue stain was then added to identify and subsequently exclude dead cells. Specifically, the cells were mixed in an approximate 1:1 ratio using trypan-blue solution. The trypan-blue was diluted to about 0.8 mM in PBS. The trypan-blue was stored at room temperature. Because all living or viable cells exclude trypan-blue, dead cells are stained blue by the dye. Accordingly, all cells stained blue were removed. Cells were suspended so that about 300 μL contained about 3×106 tumor growth cells. This concentration of cells was required for successful tumor growth at each injection site. - b.) Injection and Growth of Tumor Cells
- 1. Simultaneous with preparation of tumor growth cells, Balb/C mice had previously arrived and their health was checked.
2. All animals were allowed to acclimate for at least 72 hours.
3. All mice were about 6-8 weeks old at time of inoculation. The inoculation area was cleaned and sterilized with ethanol prior to inoculation.
4. A 1 cc syringe was filled with the cancer cells by drawing the cell mixture into the syringe without the needle. A 26-gauge needle was subsequently added to the syringe.
5. The cells were then injected subcutaneously into one lower flank of each mouse and allowed to grow until they formed a tumor which reached an average volume of about 50-60 mm3.
6. The mice were then anesthetized and the tumors were harvested by using a scalpel and appropriately stored prior to being injected into the recipient mice.
Protocol B: Insertion of Tumors from Donor Mice into Recipient Mice
21. Additional Balb/C recipient mice had previously arrived. Upon arrival of the recipient mice, the health of all mice was checked; and after passing the health test, each was numbered with a unique ear tag.
22. The recipient mice were allowed to acclimate for at least 72 hours.
23. HCT116 tumors produced in Protocol A above were removed from the donor mice by scalpel and cut into small fragments, approximately 2 mm3 in size. The 2 mm3 tumors were implanted using a 3 mm diameter trocar syringe into the right and the left flanks of each mouse (i.e., 1 tumor per flank). The tumors were permitted to grow in the recipient mice until they reached a size of about 100-200 mm3 before treatment started atday 0. Treatments continued for 21 days or until the mouse was removed from the study and euthanized or the mouse died.
24. The tumor sizes and weights of the animals were determined daily until the end of the study atday 21. -
FIGS. 38 a and 38 b show graphically the results of the oral test.FIG. 38 a shows clear difference in measured tumor volume, as a function of time, between the different compounds. The smaller the tumor, the better. Further,FIG. 38 b shows differences in mean mouse weight, as a function of time, between the different compounds. The greater the weight, the better. - Table 37 summarizes the number and the point in time during the study that the mice were removed from the study. Reasons for mice leaving the study were primarily death and large tumor size, resulting in euthanasia. The Sample ID's relate to compounds manufactured according to procedures discussed earlier herein.
-
TABLE 37 Oral Treatment No. of Mice No. of Days Group Number Sample ID Removed into Study 3 NE-28- 10X 1 11 2 14 1 15 2 Cisplatin 1 8 1 11 1 14 1 16 1 Control 1 7 2 11
Table 38 provides a comparison of the doubling time (RTV2) for each group in the study. In addition, Table 38 also lists the growth delay in days, maximum percent weight loss and statistical significance of the data. -
TABLE 38 Median Mean Time Time Growth Group to RTV2 to RTV2 Delay Maximum % Number (days) (days) (days) Significance Weight Loss 1 2.5 2.6 — — 0 2 3.9 3.5 0.9 p < 0.05 5 (d2) 3 4.0 3.7 1.1 p < 0.05 0 - This in vivo experiment was designed to determine the effects of bi-metallic Au—Pt nanocrystalline suspensions GPB-15-1, GPB-15-2 and GPB-030-1 on the behavior and quality of life in Swiss Webster mice. Specifically, female mice were given GPB-15-1 ad libitum at the start of the study (17 Jun. 2011) for 47 days. GPB-15-2 was given ad libitum for 56 days starting on 2 Aug. 2011. GPB-030-01 has been given ad libitum starting on 26 Sep. 2011 and is currently being administered. The three different bi-metallic nanocrystalline suspensions were made essentially the same way and equivalent to PGT25 herein. The female Swiss Websters have been actively drinking GPB-030-01 for 147 days as of Feb. 20, 2012. GPB-030-01 started on Sep. 26, 2011.
-
- Species: Mice
- Strain: Swiss Webster ND4
- Source: Harlan
- Gender and number: Female, 13
- Age: About 6-8 weeks old at the start of the study.
- Identification: Each mouse was given a unique identity color.
- Animal husbandry: On receipt, all animals were examined for external signs of ill-health and all unhealthy animals were excluded from further evaluation. Animals were housed in groups of 6 and 7 under normal drinking conditions, in a thermostatically monitored room (22±4° C.) in an animal unit. The health status of the animals was monitored throughout this period and the suitability of each animal for experimental use was assessed prior to study start.
- Housing Animals were housed in groups of 6 and 7 per cage in a controlled room, to ensure correct temperature, humidity and 12 hour light/dark cycle for the duration of the study on weekends. An 8 hour light and 16 hour dark during the week, Monday-Friday.
- Diet: Rodent Diet 5002 and Bottled water (such as deer park) or gold/platinum nanocrystalline suspensions are available ad libitum throughout the experimental period of the study. Only bottled water and Rodent Diet 5002 were present during the acclimatization period.
- Test gold/platinum bi-metallic nanocrystalline suspensions GPB-15-1, GPB-15-2 and GPB-030-01 (equivalent to PGT24).
- Control “
Cage 1”, Treatment “Cage 2”. The numbers of animals in each group are respectively 6 and 7.
Cage 1 (control):Day 0 Normal drinking water, given normal Rodent Diet 5002 from day 0-month 8 and present.
Cage 2 (treatment):Day 0 gold/platinum bi-metallic nanocrystalline suspension GPB-15-1 (average 4.0 ml 1 d; gold ppm: 8.6. platinum ppm: 2.3) as drinking water from day 0-day 47. GPB-15-2 (average 3.9 ml 1 d; gold ppm: 8.6: platinum ppm: 2.3) as drinking water from day 48-day 101. GPB-030-01 (average 4.3 ml 1 d; gold ppm: 8.6, platinum ppm: 2.5) as drinking water fromday 102 through 39 weeks. The mice were given normal Rodent Diet 5002 fromday 0 through 39 weeks. - On arrival of animals, the health of all animals was checked and after passing the health test, each was colored with a unique tail marking.
- The animals were allowed to acclimate for at least 1 week.
- 13 animals were purchased and separated into two ten gallon glass tanks. Seven animals were placed in a treatment group and 6 animals were placed in a control group.
- Gold/platinum bi-metallic nanocrystalline suspension were prepared so as to achieve a suspension with a concentration of about 8.6 ppm Au and 2.3 ppm Pt for GPB-15-1, 8.6 ppm Au and 2.3 ppm Pt for GPB-15-2 and 8.6 ppm Au and 2.5 ppm Pt in GPB-030-01.
- Treatments were given daily, i.e. new suspensions were replaced every 24 hours until 11 Oct. 2011. After this date, suspensions were changed every 48 hours. Samples were tested for particle size to see if there was any growth. After collecting data during the 24 hr suspension change period and no significant growth effects present, suspensions were then changed every 48 hours.
- All suspensions were administered in a glass bottle to eliminate the potential effects of plastic bottle.
- Animals were housed in a 10-gallon glass tank with a metal mesh cover. A corn cob bedding material (Bed O′ Cobs manufactured by the Andersons) was provided as a floor material, one nestlet (purchased from Ancare) was given per animal per week. Animals had access to a wheel for exercise (8 in diameter Run around wheel manufactured by Super Pet), as well as a housing unit (Pet igloo by Super Pet) and a plastic food dish (Petco plastic dish) for Certified Rodent diet.
- Cage cleaning occurred weekly where animals are housed in a plastic shoebox cage with food and drinking solution for no more than two hours.
- Each animal was weighed weekly by a calibrated balance. Balance was checked with a certified 50 g weight to insure no drifting has occurred. (Scout pro 200 g balance purchased from Fisher Scientific)
- Animal health was monitored daily
-
- 1. All animals have appeared to be in good health and are behaving normally since the study began, 17 Jun. 2011. No animals have been lost, nor removed from the study due to illness.
- 2.
FIG. 39 a shows average consumption of bi-metallic Au—Pt nanocrystalline suspensions for Cage 2 (“Treatment”) and average consumption of control drinking water in Cage 1 (“Control”) over a 39 week period. -
FIG. 39 b shows the average weight gain ofTreatment Group 2 andControl Group 1. - 3. No difference in amount of liquid consumed nor any weight gan is apparent.
- This in vitro experiment was designed to determine if nanocrystals in Au—Pt bi-metallic suspension GPB-11 could bind with genomic DNA and/or albumin; and if there was preferential binding. GPB-11 was incubated with genomic DNA from a human or a mouse, in the presence or absence of human, mouse or bovine albumin. The DNA or albumin binding to GPB-11 was characterized qualitatively and quantitatively by UV-Visible spectrophotometry.
- Albumin is a known stabilizing agent and could provide a biofunctionalized layer for water-dispersed nanoparticles. The binding affinity between gold nanoparticles and DNA has been indicated to affect DNA transcription. Albumin is also known to assist in drug delivery.
- Albumin was incubated with GPB-11 in a binding buffer at room temperature for about 1 hour to determine the differential binding of albumin to GPB-11 in the absence or presence of genomic DNA. Similarly, at the same temperature and in the same binding buffer, genomic DNA was incubated with GPB-11 for about 1 hour to measure the binding abilities of DNA to the GPB-11 when co-incubating with or without albumin. After reactions were allowed to occur, the GPB-11 suspension was spun down, washed and placed into an elution buffer for absorbance measurements.
- The binding capacities of albumin or DNA to GPB-11 were monitored by 201-UV-VIS spectrometer at A280 or A260 (e.g., λ=280 or λ=260). Absorption spectra from samples were acquired by a double beam Czerny-Turner monochromator system and dual silicon photodiodes equipped in 201-UV-VIS. The background of GPB-11, albumin and DNA were subtracted from the reaction tubes.
- Further, to visualize interactions between the DNA and GPB-11, a Fast-scan atomic force microscopy (AFM) set-up was utilized. Additionally, a nano-scale-resolution type of scanning, probe microscopy, was used to take a photomicrograph of the interaction.
-
-
Equipment and materials used for concentration Supplier Cat. No. Eppendorf centrifuge Brinkmann Instruments Inc 5417 C w Rotor Zetasizer Malvern Nano-ZS90; Model: Zen3690 1.5 ml Eppendorf Fisher Scientific 05-402-24B Tubes Pipet Tips Fisher Scientific 02-681-140 Pipetter Fisher Scientific 21-377-821 Sodium bicarbonate Fisher Scientific 144-55-8 Potassium hydroxide Fisher Scientific 1310-58-3 -
- 1. GPB-11 (having an atomic concentration of Au, 8.2 ppm; and Pt, 2.5 ppm) was placed into eppendorf tubes, and centrifuged at about 20,000×g for about 10 mins.
- 2. The pellets were clearly observed on the bottom of these tubes. The top 95% supernatant was discarded and bottom 5% supernatant and pellets were collected. The concentrated suspension was then resuspended in the binding reaction studies.
- The concentrated GPB-11 suspension was rehydrated in a solution containing 2.7 mM Sodium Hydrogen Carbonate and 2.1 mM Potassium hydroxide with the same amount as the above-described supernatant. Zeta potentials of rehydared GPB-11 and original GPB-11 solutions were measured using a Zetasizer as discusssed elswhere herein, and the results were −50.3 mV and −51.7 mV respectively. The very similar Zeta potential values suggested that rehydration of concentrated GPB-11 in the binding reaction studies should have the same effect as adding an oringinal concentration of GPB-11.
- Binding Assays of Albumin or Genomic DNA with Co-Nanocrystalline GPB-11
-
Equipment and materials used for binding assays Supplier Cat. No. 201-UV-VIS (Uvcalc-bio) Thermo Spectronic 001201 pH/Conductivity Meter Fisher Scientific Accumet AR 20; ID: I928 Vertex Mixer Fisher Scientific 02215365 Bovine serum albumin Sigma Aldrich A9418 Mouse serum albumin Sigma Aldrich A3139 Human serum albumin MP Biomedicals, 191349 LLC Human genomic DNA (female) Promega G1521 Isopropyl alcohol Sigma Aldrich W292907 Ethanol Sigma Aldrich 459836 Wizard Genomic DNA Purification Kit Promega A1120 Tris base Fisher Scientific 77-86-1 Potassium chloride (KCl) Fisher Scientific 7447-40-7 Magnesium chloride (MgCl2) Sigma Aldrich M4880 IGEPAL ® CA-630 Sigma Aldrich I8896 Hydrochloric acid Fisher Scientific 7647-01-0 Sodium hydroxide (NaOH) Fisher Scientific 1310-73-2 Ethylenediaminetetraacetic acid Acros Organics 60-00-4 (EDTA)
Isolation of Genomic DNA from Mouse Spleen and Human Whole Blood
Isolation of Genomic DNA from Mouse Spleen -
- 10 mg of thawed normal mouse spleen was added to 600 ul of chilled Nuclei Lysis Solution and incubated at 65° C. for 20 minutes.
- 3 ul of RNase Solution was put into tissue nuclei lysate, mixed and incubated at 37° C. for 25 minutes. After incubation the lysates was cooled down to room temperature.
- 200 ul of Protein Precipitation was mixed with tissue lysate, vertexed and chilled on ice for 5 minutes.
- The above mixture was centrifuged at 16000×g for 4 minutes.
- After centrifugation the supernatant was transferred to a fresh tube containing 600 ul of room temperature isopropanol and mixed gently by inversion.
- The above reactive mixture was centrifuged at 16000×g for 1 minute.
- The supernatant was removed and the pellet was resuspended in 600 ul of
room temperature 70% ethanol and centrifuged at 16000×g for 1 minute. - The ethanol was aspirated and DNA pellet was air dried for 15 minutes.
- The dried DNA pellet was rehydrated in 100 ul of DNA Rehydration Solution for overnight at 4° C.
Isolation of Genomic DNA from Human Whole Blood - 3 ml of normal human male whole blood was combined with 9 ml of Cell Lysis Solution, mixed by inversion and incubated for 10 minutes at room temperature.
- The above mixed solution was centrifuged at 2000×g for 10 minutes. The supernatant was discarded and the pellet was vertexed.
- 3 ml of Nuclei Lysis Solution was added onto the above pellet and mixed by inversion.
- 1 ml of Protein Precipitation Solution was added into the above nuclei lysate and vortexed for 20 seconds following by centrifuging at 2000×g for 10 minutes.
- After centrifugation the supernatant was transferred to a fresh tube containing 3 ml of room temperature isopropanol and mixed gently.
- The above reactive mixture was centrifuged at 2000×g for 1 minute.
- The supernatant was removed and the pellet was washed in 3 ml of
room temperature 70% ethanol and centrifuged at 2000×g for 1 minute. - The ethanol was aspirated and DNA pellet was air dried for 15 minutes.
- The dried DNA pellet was rehydrated in 250 ul of DNA Rehydration Solution for overnight at 4° C.
- The binding buffer was prepared with 20 mM Tris, 100 mM KCl, 3 mM MgCl2 and 0.1% IGEPAL. The pH was adjusted to about 7.5 by pH/Conductivity Meter with Hydrochloric acid and NaOH.
- To make 10× 50T1E (50 mM Tris-HCL/1 mM EDTA), 6.05 gram Tris base and 0.37 gram EDTA were mixed in 100 ml distilled water to dissolve. The pH of the solution was regulated to be about 8 by monitoring with a pH/Conductivity Meter and adjusting with Hydrochloric acid and NaOH. Before eluting DNA from the nanoparticles, the 10× 50T1E solution was diluted 10 times with distilled water.
-
-
TABLE 29 Groups Combinations 1 2 3 4 5 6 7 8 Albumin 0.4 mg/ml − + − + − + − + DNA 15 ug/ml − − − − + + + + GPB11 22 ug/ml − − + + − − + + Binding buffer + + + + + + + + -
- 25. The binding reaction was carried out by the incubation of GPB-11, albumin and DNA with binding buffer for about 1 hour at room temperature in eight combinations as shown in Table 29. During incubating the samples were vertexed every 5 minutes.
- 26. After incubation, the reaction solution was spun down at 20000×g for about 10 minutes at room temperature.
- 27. The pellets were washed once and resuspened in 400 ul DNA elution buffer.
- 28. The absorbance at 280 nm for albumin (i.e., absorption peak) and 260 nm for DNA (i.e., absorption peak) was measured with 201-UV-VIS.
-
-
Equipment and materials used for imaging Supplier Cat. No. Dimension FastScan AFM Bruker system FastScan A probe AppNano Probe model: UHF Series Mica Bruker Spin Coater Instras Scientific SCK-100 - After the binding reaction was permitted to occur, 50 ul of the mixture of human female genomic DNA and GPB-11 in binding buffer was deposited and spin-coated (at least 3000 rpm) onto a fresh mica sheet. The mica-containing sample was rinsed with clean water once, followed by drying in air. Imaging was carried out by FastScan AFM with NanoScope V and Stage Controller. The AFM was operated in tapping mode and FastScan A probe (k˜17 N/m) was used. High resolution phase mapping, overlaying topography (3D) and height in cross sections were analyzed by FastScan NanoScope Software. Results are discussed later herein.
- The absorbance of albumin binding to GPB-11 was measured at 280 nm. Different combinations of albumin and GPB-11 were tested in the presence or absence of genomic DNA. Table 30 shows that very similar results were achieved among different albumin and GPB11 combinations. Representative data are also depicted in
FIG. 40 a . -
TABLE 30 Experiments Combinations 1 2 3 4 5 6 Albumin Bovine + − − + − − Mouse − + − − + − Human − − + − − + Genomic DNA Mouse − − − + + − Human − − − − − + GPB-11 + + + + + + - Specifically,
FIG. 40 a shows graphically the amount of mouse albumin binding in the presence or absence of mouse genomic DNA as a function of the absorbance at 280 nm. In the absence of genomic DNA, albumin significantly bound to the bi-metallic nanocrystals in GPB-11. But when genomic DNA was added in binding assay, no albumin binding to the nanocrystals in GPB-11 was observed. These results indicated that the nanocrystals in GPB-11 can bind with albumin, but preferentially binds to mouse genome DNA. In another words, the Au—Pt bi-metallic nanocrystals in GPB-11 apparently have a soft corona of albumin. - DNA binding to nanocrystals in GPB-11 was determined by measuring the absorbance at 260 nm. The binding ability of mouse or human genomic DNA to bi-metallic nanocrystals in GPB-11 was measured with different combinations of albumin. Table 31 shows the various combinations or mixtures tested. Highly consistent results were observed between different DNA and nanocrystals in GPB-11 combinations. The representative results are depicted graphically in
FIG. 40 b . -
TABLE 31 Experiments Combinations 1 2 3 4 5 Genomic DNA Mouse + − + − + Human − + − + − Albumin Bovine − − + − − Mouse − − − − + Human − − − + − GPB-11 + + + + + - Specifically,
FIG. 40 b shows graphically the amount of DNA binding in the presence or absence of mouse albumin.FIG. 40 b shows that in both, the presence and the absence of albumin, genomic DNA significantly bound to nanocrystals in GPB-11. When albumin was absent, the amount of DNA binding with GPB-11 nanocrystals was dramatic. Even when a large amount of albumin was added in the binding assay, a statistically significant amount of DNA was observed to be bound to the GPB-11 bi-metallic nanocrystals. These results further confirm that bi-metallic nanocrystals in GPB-11 bind to genomic DNA much stronger than albumin. Further, without wishing to be bound by any particular theory or explanation, it is possible that the Au—Pt bi-metallic nanocrystals in GPB-11 may bind to genomic DNA (when in the presence thereof) with covalent bonds. Such bonding could affect DNA function. - An attempt was made to image DNA binding to Au—Pt bi-metallic nanocrystals. Specifically, the samples in DNA binding assay were imaged by an AFM. A representative result is shown in
FIG. 40 c . It is clearly shown that Au—Pt bi-metallic nanocrystals bound to human genomic DNA. Most nanocrystals were observed binding on the end of string DNA molecules. The diameters of the imaged nanoparticles are within the size range of the nanocrystals in GPB-11, thus confirming the binding.
Claims (20)
1. A pharmaceutically acceptable suspension comprising:
a.) pharmaceutical grade water;
b.) at least one processing enhancer; and
c.) gold-platinum bi-metallic nanocrystals suspended in said water forming a suspension, wherein said gold-platinum bi-metallic nanocrystals:
i.) have surfaces that include at least one characteristic selected from the group of characteristics consisting of: (1) no organic chemical constituents adhered or attached to said surfaces and (2) are substantially clean and do not have chemical constituents adhered or attached to surfaces, other than water, lysis products of water or said processing enhancer, none of which alter the functioning of said nanocrystals;
ii.) have a particle size of less than about 50 nm;
iii.) are present in said suspension at a total atomic metal concentration of about 2-1000 ppm; and
d.) said suspension having a pH of between about 5 to about 12 and a zeta potential of at least about −30 mv.
2. The composition of claim 1 , wherein said processing enhancer comprises sodium bicarbonate.
3. The composition of claim 1 , wherein said suspension has a zeta potential of at least about −40 mV and a pH of between about 8 to about 12.
4. The composition of claim 1 , wherein said suspension has a zeta potential of at least about −50 mV.
5. The composition of claim 1 , wherein said surfaces have no organic chemical constituents adhered or attached to said surfaces.
6. The composition of claim 1 , wherein said surfaces are substantially clean and do not have chemical constituents adhered or attached to surfaces, other than lysis products of said water.
7. The composition of claim 1 , wherein said suspension has a total metal concentration of about 10-500 ppm.
8. The composition of claim 1 , wherein said gold-platinum bi-metallic nanocrystals comprise an alloy of gold and platinum.
9. The composition of claim 8 , wherein platinum is a minor constituent in said bi-metallic nanocrystals and gold is a major constituent in said bi-metallic nanocrystals.
10. The composition of claim 1 , wherein said suspension is free from chlorides and chlorine-based species.
11. A pharmaceutical suspension comprising:
a.) pharmaceutical grade water containing at least one processing enhancer, said water having a pH of between about 5 to about 12;
b.) gold-platinum bi-metallic alloyed nanocrystals in said water forming said suspension, said suspension having a zeta potential of at least about −30 mV and wherein said gold-platinum bi-metallic alloyed nanocrystals:
i.) have surfaces that include at least one characteristic selected from the group of characteristics consisting of: (1) no organic chemical constituents adhered or attached to said surfaces and (2) are substantially clean and do not have chemical constituents adhered or attached to surfaces thereof;
ii.) have an average particle size of less than about 50 nm; and
iii.) are present in said suspension at a concentration of about 2-1000 ppm.
12. The composition of claim 11 , wherein said suspension has a zeta potential of at least about −40 mV and a pH of between about 8 to about 12.
13. The composition of claim 11 , wherein said suspension is free from chlorides and chlorine-based species.
14. The composition of claim 11 , wherein said surfaces are substantially clean and do not have chemical constituents adhered or attached to surfaces, other than water or lysis products of water and said suspension is free from chlorides and chlorine-based species.
15. The composition of claim 11 , wherein at least some platinum ions are present in said water suspension.
16. A suspension comprising:
a.) pure water containing at least one processing enhancer, said water having a pH of between about 5 to about 12;
b.) gold-platinum bi-metallic nanocrystals in said water forming said suspension, said suspension having a zeta potential of at least about −30 mV and wherein said gold-platinum nanocrystals:
i.) have surfaces that include at least one characteristic selected from the group of characteristics consisting of: (1) no organic chemical constituents adhered or attached to said surfaces and (2) are substantially clean and do not have chemical constituents adhered or attached to surfaces, other than water, lysis products of water or said processing enhancer, none of which alter catalytic functioning of said nanocrystals;
ii.) have a particle size of less than about 50 nm; and
iii.) are present in said suspension at a concentration of about 2-1000 ppm.
17. The composition of claim 16 , wherein said suspension has a zeta potential of at least about −40 mV and a pH of between about 8 to about 12.
18. The composition of claim 17 , wherein said surfaces have no organic chemical constituents adhered or attached to said surfaces.
19. The composition of claim 16 , wherein said surfaces are substantially clean and do not have chemical constituents adhered or attached to surfaces, other than water or lysis products of water.
20. The composition of claim 19 , wherein said suspension does not contain any chlorides or chlorine-based materials used to form the gold-platinum bi-metallic nanocrystals.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/708,253 US20230172975A1 (en) | 2011-03-30 | 2022-03-30 | Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same |
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201161469525P | 2011-03-30 | 2011-03-30 | |
PCT/US2012/031654 WO2012135743A2 (en) | 2011-03-30 | 2012-03-30 | Novel gold-platinum based bi-metallic nanocrystal suspensions, electrochemical manufacturing processes therefor and uses for the same |
US201314008931A | 2013-12-16 | 2013-12-16 | |
US15/204,534 US20160317578A1 (en) | 2011-03-30 | 2016-07-07 | Novel Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same |
US17/708,253 US20230172975A1 (en) | 2011-03-30 | 2022-03-30 | Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US15/204,534 Division US20160317578A1 (en) | 2011-03-30 | 2016-07-07 | Novel Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same |
Publications (1)
Publication Number | Publication Date |
---|---|
US20230172975A1 true US20230172975A1 (en) | 2023-06-08 |
Family
ID=46932416
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/008,931 Active US9387225B2 (en) | 2011-03-30 | 2012-03-30 | Gold-platinum based bi-metallic nanocrystal suspensions, electrochemical manufacturing processes therefor and uses for the same |
US15/204,534 Abandoned US20160317578A1 (en) | 2011-03-30 | 2016-07-07 | Novel Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same |
US17/708,253 Abandoned US20230172975A1 (en) | 2011-03-30 | 2022-03-30 | Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/008,931 Active US9387225B2 (en) | 2011-03-30 | 2012-03-30 | Gold-platinum based bi-metallic nanocrystal suspensions, electrochemical manufacturing processes therefor and uses for the same |
US15/204,534 Abandoned US20160317578A1 (en) | 2011-03-30 | 2016-07-07 | Novel Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same |
Country Status (14)
Country | Link |
---|---|
US (3) | US9387225B2 (en) |
EP (1) | EP2691082B1 (en) |
JP (1) | JP6121985B2 (en) |
KR (2) | KR101955735B1 (en) |
CN (1) | CN103764123B (en) |
BR (1) | BR112013025112A2 (en) |
CA (1) | CA2829095C (en) |
DK (1) | DK2691082T3 (en) |
ES (1) | ES2564672T3 (en) |
IL (1) | IL228356A (en) |
MX (1) | MX337439B (en) |
RU (1) | RU2617055C2 (en) |
SG (1) | SG193439A1 (en) |
WO (1) | WO2012135743A2 (en) |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3889102A3 (en) * | 2007-07-11 | 2022-01-05 | Clene Nanomedicine, Inc. | Continuous methods for treating liquids and manufacturing certain constituents (e.g., nanoparticles) in liquids, apparatuses and nanoparticles and nanoparticle/liquid solution(s) resulting therefrom |
EP2735389A1 (en) * | 2012-11-23 | 2014-05-28 | Universität Duisburg-Essen | Process for the preparation of pure, in particular carbon-free nanoparticles |
JP5891320B1 (en) * | 2015-02-12 | 2016-03-22 | 秋田県 | Processing method using zeta potential control method |
CN106680450B (en) * | 2017-03-23 | 2023-04-25 | 江西省农业科学院农业经济与信息研究所 | Water quality monitoring device and method |
US11674234B1 (en) * | 2020-09-30 | 2023-06-13 | National Technology & Engineering Solutions Of Sandia, Llc | Electrodeposited platinum-gold alloy |
CN112683710B (en) * | 2020-12-02 | 2024-04-16 | 大连理工大学 | Test device for accelerated corrosion of reinforced concrete bridge plate under chloride ion corrosion |
CN112935273A (en) * | 2021-01-26 | 2021-06-11 | 哈尔滨理工大学 | Method for preparing CuPt alloy nanoparticles at room temperature |
JP2022131579A (en) | 2021-02-26 | 2022-09-07 | キオクシア株式会社 | Analysis device and analysis method |
CN113500199B (en) * | 2021-06-10 | 2022-11-08 | 浙江大学 | Preparation method of gold-platinum-based bimetallic active oxygen self-generating nano material, product and application thereof |
CN113777034B (en) * | 2021-08-20 | 2024-04-19 | 嘉兴学院 | Gold nano bipyramid array substrate and preparation method and application thereof |
CN114470201B (en) * | 2021-12-24 | 2024-03-19 | 上海市儿童医院 | Gold-platinum nanoparticle loaded with KCNA5 antibody and preparation method and application thereof |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20030099718A1 (en) * | 2001-04-23 | 2003-05-29 | Burrell Robert Edward | Treatment of mucosal membranes |
WO2010083040A1 (en) * | 2009-01-15 | 2010-07-22 | Gr Intellectual Reserve, Llc | Continuous semicontinuous and batch methods for treating liquids and manufacturing certain constituents (e.g., nanoparticles) in liquids, apparatuses and nanoparticles and nanoparticle/liquid solution(s) and colloids resulting therefrom |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU3679801A (en) * | 2000-02-08 | 2001-08-20 | Rice University | Optically-active nanoparticles for use in therapeutic and diagnostic methods |
US6989157B2 (en) * | 2000-07-27 | 2006-01-24 | Nucryst Pharmaceuticals Corp. | Dry powders of metal-containing compounds |
US7001617B2 (en) * | 2001-04-23 | 2006-02-21 | Nueryst Pharmaceuticals Corp. | Method of induction of apoptosis and inhibition of matrix metalloproteinases using antimicrobial metals |
JP2002212102A (en) * | 2001-01-23 | 2002-07-31 | Ainobekkusu Kk | Electrochemically bioactive fine particle |
US20080190770A1 (en) * | 2004-04-26 | 2008-08-14 | Cap Technologies, Llc | Treatment of Fluids and/or Sludge with Electro Plasma |
TW200704404A (en) * | 2005-03-23 | 2007-02-01 | Shetech Co Ltd | Precious metal nanocolloid solution |
EP1976618A2 (en) * | 2006-01-03 | 2008-10-08 | GR Intellectual Reserve, LLC | Methods and apparatuses for making liquids more reactive |
KR100857389B1 (en) * | 2006-06-30 | 2008-09-11 | (주)아모레퍼시픽 | AP-GRR peptide or peptide chain containing AP-GRR peptide, and drug-delivery carrier comprising the same |
TW200819540A (en) * | 2006-07-11 | 2008-05-01 | Genelux Corp | Methods and compositions for detection of microorganisms and cells and treatment of diseases and disorders |
EP3889102A3 (en) | 2007-07-11 | 2022-01-05 | Clene Nanomedicine, Inc. | Continuous methods for treating liquids and manufacturing certain constituents (e.g., nanoparticles) in liquids, apparatuses and nanoparticles and nanoparticle/liquid solution(s) resulting therefrom |
EP2200932A4 (en) * | 2007-09-21 | 2014-09-10 | Cytimmune Sciences Inc | Nanotherapeutic colloidal metal compositions and methods |
KR102051248B1 (en) * | 2009-07-08 | 2019-12-02 | 클레네 나노메디슨, 인크. | Novel gold-based nanocrystals for medical treatments and electrochemical manufacturing processes therefor |
KR101090431B1 (en) * | 2009-08-19 | 2011-12-07 | 삼성전기주식회사 | Method for preparing metal nanoparticles using matal seed and metal nanoparticles comprising metal seed |
-
2012
- 2012-03-30 ES ES12764547.1T patent/ES2564672T3/en active Active
- 2012-03-30 US US14/008,931 patent/US9387225B2/en active Active
- 2012-03-30 KR KR1020137028744A patent/KR101955735B1/en active IP Right Grant
- 2012-03-30 CA CA2829095A patent/CA2829095C/en active Active
- 2012-03-30 JP JP2014502869A patent/JP6121985B2/en active Active
- 2012-03-30 MX MX2013011245A patent/MX337439B/en active IP Right Grant
- 2012-03-30 RU RU2013148011A patent/RU2617055C2/en active
- 2012-03-30 KR KR1020197006058A patent/KR102103534B1/en active IP Right Grant
- 2012-03-30 DK DK12764547.1T patent/DK2691082T3/en active
- 2012-03-30 SG SG2013068861A patent/SG193439A1/en unknown
- 2012-03-30 BR BR112013025112-3A patent/BR112013025112A2/en not_active Application Discontinuation
- 2012-03-30 WO PCT/US2012/031654 patent/WO2012135743A2/en active Application Filing
- 2012-03-30 CN CN201280016524.9A patent/CN103764123B/en active Active
- 2012-03-30 EP EP12764547.1A patent/EP2691082B1/en active Active
-
2013
- 2013-09-11 IL IL228356A patent/IL228356A/en active IP Right Grant
-
2016
- 2016-07-07 US US15/204,534 patent/US20160317578A1/en not_active Abandoned
-
2022
- 2022-03-30 US US17/708,253 patent/US20230172975A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20030099718A1 (en) * | 2001-04-23 | 2003-05-29 | Burrell Robert Edward | Treatment of mucosal membranes |
WO2010083040A1 (en) * | 2009-01-15 | 2010-07-22 | Gr Intellectual Reserve, Llc | Continuous semicontinuous and batch methods for treating liquids and manufacturing certain constituents (e.g., nanoparticles) in liquids, apparatuses and nanoparticles and nanoparticle/liquid solution(s) and colloids resulting therefrom |
Also Published As
Publication number | Publication date |
---|---|
RU2617055C2 (en) | 2017-04-19 |
BR112013025112A2 (en) | 2020-09-29 |
US20140294963A1 (en) | 2014-10-02 |
IL228356A (en) | 2017-04-30 |
KR20140020310A (en) | 2014-02-18 |
SG193439A1 (en) | 2013-10-30 |
CA2829095A1 (en) | 2012-10-04 |
EP2691082B1 (en) | 2016-01-13 |
NZ617018A (en) | 2015-10-30 |
RU2013148011A (en) | 2015-05-10 |
MX2013011245A (en) | 2013-10-17 |
DK2691082T3 (en) | 2016-02-29 |
WO2012135743A3 (en) | 2014-05-01 |
KR20190027933A (en) | 2019-03-15 |
JP6121985B2 (en) | 2017-04-26 |
AU2012236213A2 (en) | 2014-01-23 |
IL228356A0 (en) | 2013-12-31 |
EP2691082A2 (en) | 2014-02-05 |
KR101955735B1 (en) | 2019-03-07 |
CA2829095C (en) | 2019-08-27 |
KR102103534B1 (en) | 2020-04-23 |
US9387225B2 (en) | 2016-07-12 |
EP2691082A4 (en) | 2015-06-24 |
CN103764123A (en) | 2014-04-30 |
AU2012236213A1 (en) | 2013-11-14 |
WO2012135743A2 (en) | 2012-10-04 |
US20160317578A1 (en) | 2016-11-03 |
MX337439B (en) | 2016-03-03 |
ES2564672T3 (en) | 2016-03-28 |
JP2014518847A (en) | 2014-08-07 |
CN103764123B (en) | 2018-03-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20230172975A1 (en) | Gold-Platinum Based Bi-Metallic Nanocrystal Suspensions, Electrochemical Manufacturing Processes Therefor and Uses for the Same | |
JP6703052B2 (en) | Novel gold-based nanocrystal for medical treatment, and electrochemical production method for the gold-based nanocrystal | |
AU2019204420B2 (en) | Novel gold-based nanocrystals | |
Kim et al. | Synthesis of hex nut shaped Au–Ag nanostructures via a galvanic replacement reaction and their optical properties | |
AU2012236213B2 (en) | Novel gold-platinum based bi-metallic nanocrystal suspensions, electrochemical manufacturing processes therefor and uses for the same | |
NZ617018B2 (en) | Novel gold-platinum based bi-metallic nanocrystal suspensions, electrochemical manufacturing processes therefor and uses for the same | |
Deng | Sputter deposition and formation mechanism of Pt and Pt alloy nanoparticles |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |