CN104549182A - Branched boric acid functionalized monolithic column and preparation method and application thereof - Google Patents
Branched boric acid functionalized monolithic column and preparation method and application thereof Download PDFInfo
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- CN104549182A CN104549182A CN201410856918.6A CN201410856918A CN104549182A CN 104549182 A CN104549182 A CN 104549182A CN 201410856918 A CN201410856918 A CN 201410856918A CN 104549182 A CN104549182 A CN 104549182A
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- boric acid
- integral post
- dendroid
- flexible
- acid functionalization
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- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 title claims abstract description 124
- 239000004327 boric acid Substances 0.000 title claims abstract description 108
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 238000007306 functionalization reaction Methods 0.000 claims abstract description 96
- 238000000926 separation method Methods 0.000 claims abstract description 9
- 235000010338 boric acid Nutrition 0.000 claims description 118
- 229960002645 boric acid Drugs 0.000 claims description 118
- 241000555268 Dendroides Species 0.000 claims description 80
- 108090000288 Glycoproteins Proteins 0.000 claims description 44
- 102000003886 Glycoproteins Human genes 0.000 claims description 44
- 238000006243 chemical reaction Methods 0.000 claims description 30
- 239000000412 dendrimer Substances 0.000 claims description 29
- 229920000736 dendritic polymer Polymers 0.000 claims description 29
- 239000004593 Epoxy Substances 0.000 claims description 26
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 23
- 238000006011 modification reaction Methods 0.000 claims description 20
- 239000000178 monomer Substances 0.000 claims description 16
- -1 saccharide compound Chemical class 0.000 claims description 15
- OZAIFHULBGXAKX-UHFFFAOYSA-N 2-(2-cyanopropan-2-yldiazenyl)-2-methylpropanenitrile Chemical compound N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 claims description 13
- 230000035484 reaction time Effects 0.000 claims description 12
- 239000003431 cross linking reagent Substances 0.000 claims description 11
- 239000004088 foaming agent Substances 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 11
- 229920000620 organic polymer Polymers 0.000 claims description 11
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 claims description 10
- 229960001760 dimethyl sulfoxide Drugs 0.000 claims description 8
- LQZZUXJYWNFBMV-UHFFFAOYSA-N dodecan-1-ol Chemical compound CCCCCCCCCCCCO LQZZUXJYWNFBMV-UHFFFAOYSA-N 0.000 claims description 8
- 238000006116 polymerization reaction Methods 0.000 claims description 7
- DBCAQXHNJOFNGC-UHFFFAOYSA-N 4-bromo-1,1,1-trifluorobutane Chemical compound FC(F)(F)CCCBr DBCAQXHNJOFNGC-UHFFFAOYSA-N 0.000 claims description 5
- 230000015572 biosynthetic process Effects 0.000 claims description 5
- STVZJERGLQHEKB-UHFFFAOYSA-N ethylene glycol dimethacrylate Substances CC(=C)C(=O)OCCOC(=O)C(C)=C STVZJERGLQHEKB-UHFFFAOYSA-N 0.000 claims description 5
- 125000000524 functional group Chemical group 0.000 claims description 5
- 238000003786 synthesis reaction Methods 0.000 claims description 5
- 125000004356 hydroxy functional group Chemical group O* 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- KWNPRVWFJOSGMZ-UHFFFAOYSA-N 2-boronobenzoic acid Chemical compound OB(O)C1=CC=CC=C1C(O)=O KWNPRVWFJOSGMZ-UHFFFAOYSA-N 0.000 claims description 2
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical group [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 claims description 2
- 239000002202 Polyethylene glycol Substances 0.000 claims description 2
- 125000003368 amide group Chemical group 0.000 claims description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 2
- 125000004386 diacrylate group Chemical group 0.000 claims description 2
- KFIFDKLIFPYSAZ-UHFFFAOYSA-N formyloxy(phenyl)borinic acid Chemical compound O=COB(O)C1=CC=CC=C1 KFIFDKLIFPYSAZ-UHFFFAOYSA-N 0.000 claims description 2
- 238000002513 implantation Methods 0.000 claims description 2
- 229920001223 polyethylene glycol Polymers 0.000 claims description 2
- 125000003396 thiol group Chemical group [H]S* 0.000 claims description 2
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 claims 1
- 238000004458 analytical method Methods 0.000 abstract description 11
- 229920002521 macromolecule Polymers 0.000 abstract description 2
- 230000003044 adaptive effect Effects 0.000 abstract 1
- 238000010979 pH adjustment Methods 0.000 abstract 1
- 238000000746 purification Methods 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 57
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 32
- 239000000463 material Substances 0.000 description 27
- 239000000523 sample Substances 0.000 description 27
- 229920002873 Polyethylenimine Polymers 0.000 description 21
- 210000003296 saliva Anatomy 0.000 description 18
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 16
- 229960005305 adenosine Drugs 0.000 description 16
- 241000208125 Nicotiana Species 0.000 description 15
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 15
- 239000000243 solution Substances 0.000 description 14
- 239000002585 base Substances 0.000 description 13
- YCIMNLLNPGFGHC-UHFFFAOYSA-N catechol Chemical compound OC1=CC=CC=C1O YCIMNLLNPGFGHC-UHFFFAOYSA-N 0.000 description 12
- 239000000203 mixture Substances 0.000 description 12
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 11
- 108010066476 ribonuclease B Proteins 0.000 description 11
- 229910052796 boron Inorganic materials 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 description 9
- 229920006395 saturated elastomer Polymers 0.000 description 9
- 238000001179 sorption measurement Methods 0.000 description 9
- PXFBZOLANLWPMH-UHFFFAOYSA-N 16-Epiaffinine Natural products C1C(C2=CC=CC=C2N2)=C2C(=O)CC2C(=CC)CN(C)C1C2CO PXFBZOLANLWPMH-UHFFFAOYSA-N 0.000 description 8
- 238000010494 dissociation reaction Methods 0.000 description 8
- 230000005593 dissociations Effects 0.000 description 8
- 239000002253 acid Substances 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 108010015865 Transferrins Proteins 0.000 description 6
- 102000002070 Transferrins Human genes 0.000 description 6
- 230000004913 activation Effects 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 238000011835 investigation Methods 0.000 description 6
- 239000012160 loading buffer Substances 0.000 description 6
- 239000003960 organic solvent Substances 0.000 description 6
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 5
- 229910019142 PO4 Inorganic materials 0.000 description 5
- 239000007853 buffer solution Substances 0.000 description 5
- 235000019504 cigarettes Nutrition 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 5
- 239000003480 eluent Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 5
- 239000010452 phosphate Substances 0.000 description 5
- 239000012488 sample solution Substances 0.000 description 5
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 4
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 4
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 4
- 239000003513 alkali Substances 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000003638 chemical reducing agent Substances 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000001962 electrophoresis Methods 0.000 description 4
- 150000002148 esters Chemical class 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 3
- 239000005695 Ammonium acetate Substances 0.000 description 3
- 235000019257 ammonium acetate Nutrition 0.000 description 3
- 229940043376 ammonium acetate Drugs 0.000 description 3
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 3
- XSMJZKTTXZAXHD-UHFFFAOYSA-N ethene;2-methylprop-2-enoic acid Chemical group C=C.CC(=C)C(O)=O XSMJZKTTXZAXHD-UHFFFAOYSA-N 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 230000013595 glycosylation Effects 0.000 description 3
- 238000006206 glycosylation reaction Methods 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000007142 ring opening reaction Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- PVAWONNALJEQJH-UHFFFAOYSA-N (2,4-difluoro-3-formylphenyl)boronic acid Chemical class OB(O)C1=CC=C(F)C(C=O)=C1F PVAWONNALJEQJH-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 229910021538 borax Inorganic materials 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 230000000269 nucleophilic effect Effects 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 229920001296 polysiloxane Polymers 0.000 description 2
- 150000003141 primary amines Chemical class 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 239000004328 sodium tetraborate Substances 0.000 description 2
- 235000010339 sodium tetraborate Nutrition 0.000 description 2
- 238000010183 spectrum analysis Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000002137 ultrasound extraction Methods 0.000 description 2
- YJXJCPZHXDDRNH-UHFFFAOYSA-N (2-fluoro-3-formylphenyl)boronic acid Chemical compound OB(O)C1=CC=CC(C=O)=C1F YJXJCPZHXDDRNH-UHFFFAOYSA-N 0.000 description 1
- DQJCDTNMLBYVAY-ZXXIYAEKSA-N (2S,5R,10R,13R)-16-{[(2R,3S,4R,5R)-3-{[(2S,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}-5-(ethylamino)-6-hydroxy-2-(hydroxymethyl)oxan-4-yl]oxy}-5-(4-aminobutyl)-10-carbamoyl-2,13-dimethyl-4,7,12,15-tetraoxo-3,6,11,14-tetraazaheptadecan-1-oic acid Chemical compound NCCCC[C@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)CC[C@H](C(N)=O)NC(=O)[C@@H](C)NC(=O)C(C)O[C@@H]1[C@@H](NCC)C(O)O[C@H](CO)[C@H]1O[C@H]1[C@H](NC(C)=O)[C@@H](O)[C@H](O)[C@@H](CO)O1 DQJCDTNMLBYVAY-ZXXIYAEKSA-N 0.000 description 1
- 108010008629 CA-125 Antigen Proteins 0.000 description 1
- 102000007269 CA-125 Antigen Human genes 0.000 description 1
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 1
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
- 108010015899 Glycopeptides Proteins 0.000 description 1
- 102000002068 Glycopeptides Human genes 0.000 description 1
- 101710084021 Large envelope protein Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000002262 Schiff base Substances 0.000 description 1
- 150000004753 Schiff bases Chemical class 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- LPQOADBMXVRBNX-UHFFFAOYSA-N ac1ldcw0 Chemical compound Cl.C1CN(C)CCN1C1=C(F)C=C2C(=O)C(C(O)=O)=CN3CCSC1=C32 LPQOADBMXVRBNX-UHFFFAOYSA-N 0.000 description 1
- 238000007259 addition reaction Methods 0.000 description 1
- 150000001299 aldehydes Chemical group 0.000 description 1
- 102000013529 alpha-Fetoproteins Human genes 0.000 description 1
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229940125773 compound 10 Drugs 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000018823 dietary intake Nutrition 0.000 description 1
- 238000005906 dihydroxylation reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 125000006575 electron-withdrawing group Chemical group 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 125000003700 epoxy group Chemical group 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000003546 flue gas Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- ZLVXBBHTMQJRSX-VMGNSXQWSA-N jdtic Chemical compound C1([C@]2(C)CCN(C[C@@H]2C)C[C@H](C(C)C)NC(=O)[C@@H]2NCC3=CC(O)=CC=C3C2)=CC=CC(O)=C1 ZLVXBBHTMQJRSX-VMGNSXQWSA-N 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000003808 methanol extraction Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 229920000671 polyethylene glycol diacrylate Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- ATCZPWFDFFKCPF-UHFFFAOYSA-K trisodium;acetic acid;phosphate Chemical compound [Na+].[Na+].[Na+].CC(O)=O.[O-]P([O-])([O-])=O ATCZPWFDFFKCPF-UHFFFAOYSA-K 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/282—Porous sorbents
- B01J20/285—Porous sorbents based on polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/22—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the construction of the column
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/3085—Chemical treatments not covered by groups B01J20/3007 - B01J20/3078
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2220/00—Aspects relating to sorbent materials
- B01J2220/50—Aspects relating to the use of sorbent or filter aid materials
- B01J2220/54—Sorbents specially adapted for analytical or investigative chromatography
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention provides a branched boric acid functionalized monolithic column which is a flexible branched boric acid functionalized monolithic column obtained by bonding flexible branched macromolecules onto the hole surface of a base monolithic column through the replacement of boric acid functionalization. The invention further provides a preparation method of the branched boric acid functionalized monolithic column and the application thereof in separation, enrichment and purification of biomolecules containing cis-dyhydroxyl structures. According to the branched boric acid functionalized monolithic column and the preparation method and the application thereof, the monolithic column is high in affinity and capacity, wide in adaptive PH range, high in capacity of resisting disturbance and being suitable for direct analysis of practical samples without PH adjustment.
Description
Technical field
The invention belongs to functionalization material apparatus technical field, relate to a kind of dendroid boric acid functionalization integral post and its preparation method and application, be specifically related to high-affinity, high power capacity flexible dendroid boric acid functionalization integral post preparation and in complex sample to containing the application in being separated of biomolecule (as low-abundance glycoprotein and saccharide compound etc.) of cis-form dihydroxy structure, enrichment and purifying.
Background technology
In the research object that proteomics, metabolism group and sugar group are learned, the class biomolecule containing cis-form dihydroxy structure such as glycopeptide and glycoprotein, nucleosides and nucleotides and monose and polysaccharide are the important biomolecule of a class.These cis-form dihydroxy biomolecule play extremely important effect in physiology course and disease early diagnosis.Glycoprotein plays an important role in molecular recognition, cell and in intercellular signal conduction and immune response etc., some glycoprotein are the biomarker of cancer and the target for the treatment of clinically, as alpha-fetoprotein, carcinomebryonic antigen, CA125 and PSA etc.But glycoprotein abundance is in the sample general all very low and be subject to the strong interference of high abundance component, and therefore carrying out selective separation enrichment to it is the key detected these glycoprotein assay.In addition, saccharide compound has material impact to the quality of tobacco leaf and flue gas (as color and luster, elasticity, jealous, fragrance and tar growing amount etc.), and the distribution of the carbohydrate accurately understood in tobacco leaf and cigarette is research and development and production high-quality cigarette and personalized cigarette, the important evidence dissecting domestic and international cigarette formula.Therefore, the analytical method developing glucide in tobacco accurately and reliably has important function.
Technology that boron is affine is the class unique method of Selective recognition along dihydroxy compounds, under higher pH condition, boric acid aglucon energy and form cyclic ester along the o-dihydroxy covalent bond in dihydroxy molecule, this ester can dissociate in acid condition, thus discharge along dihydroxy molecule, reach the effect of effective separation and concentration.Although substituted boracic acid has shown wide application prospect in selective enrichment cis-form dihydroxy compound, also there is affinity shortcoming strong not in boric acid, the dissociation constant of most of substituted boracic acid and cis-form dihydroxy compound 10 at present
-1to 10
-4between [see Tetrahedron 2004,60,11205 – 11209; Anal.Chem.2013,85,2361 – 2369], therefore conventional boron affinitive material at the very low cis-form dihydroxy compound of enriched concentration as still there is sizable challenge in some glycoprotein and glycan etc.In order to address this problem, we have developed a kind of dendroid boric acid functional magnetic nano material [see Chem.Sci.2013,4,4298-4303].But adhesion reaches far away conventional antibody horizontal, can not directly use under neutrality or weakly acidic condition, so the boric acid functionalization material further developing more high-affinity under neutrality or solutions of weak acidity is vital, is worth further investigation and inquires into.
Summary of the invention
The shortcoming of prior art in view of the above, namely in order to overcome the weak shortcoming of existing boric acid functionalization material affinity, the object of the invention is to utilize nano pore to improve synergistic principle and prepare a kind of novel flexible dendroid boric acid functionalization integral post, this dendroid boric acid functionalization integral post is as a kind of high-affinity, the flexible dendroid boric acid functionalization integral post of high power capacity, can directly be separated the biomolecule (as low-abundance glycoprotein and saccharide compound etc.) containing cis-form dihydroxy structure in complex sample, the application such as enrichment and purifying.
For achieving the above object and other relevant objects, the invention provides a kind of dendroid boric acid functionalization integral post, it is the flexible dendroid boric acid functionalization integral post obtained through substituted boracic acid functionalization after the flexible dendrimer of base integral post holes surface bond.
Preferably, described base integral post is Organic Polymer Monolithic Columns.Preferably, described Organic Polymer Monolithic Columns be selected from Epoxy functionalized Organic Polymer Monolithic Columns or other contain in the Organic Polymer Monolithic Columns of active group any one.
Preferred, the described Organic Polymer Monolithic Columns that other contain active group is selected from carboxyl-functional, mercapto-functionalized, amino functional, any one in the integral post of hydroxy functionalized and aldehyde radical functionalization.
Preferred, described Organic Polymer Monolithic Columns is Epoxy functionalized Organic Polymer Monolithic Columns.
Preferably, described flexible dendrimer is selected from any one in polymine-dendrimer or other dendrimer.
Preferably, described flexible dendrimer is polymine-dendrimer (PEI).
Preferred, the molecular weight of described polymine-dendrimer be selected from 600,1800,10000 and 70000 any one.
Preferably, described substituted boracic acid is selected from any one in formylphenylboronic acid or Carboxybenzeneboronic acid.Described substituted boracic acid is containing strong electron-withdrawing group group.
Preferably, described substituted boracic acid is the fluoro-3-formylphenylboronic acid (DFFPBA) of 2,4-bis-.
Preferably, the reducing agent of described substituted boracic acid is sodium cyanoborohydride (NaCNBH
3).
Because flexible dendrimer contains a large amount of surface functional groups, after boric acid modified, integral post hole surface contains highdensity boric acid base group, the collaborative combination of multivalence can be produced to biomolecule such as glycoprotein and the glycan etc. being rich in cis-form dihydroxy, thus significantly improve active force, the binding capacity of small molecular sugar can be improved simultaneously.
The invention also discloses a kind of preparation method of dendroid boric acid functionalization integral post, specifically comprise the following steps:
1) obtain pre-polymerization liquid after the monomer containing active group, crosslinking agent, pore-foaming agent and initator being mixed, be filled in capillary, carry out polymerisation, synthesis obtains the base integral post with active group;
Preferably, the described monomer containing active group is selected from containing epoxy functionality, carboxyl functional group, mercapto functional group, amido functional group, any one in the monomer of hydroxy functional group and aldehyde functions.
Preferably, the described monomer containing active group is the monomer containing epoxy functionality.
Preferred, the described monomer containing epoxy functionality is GMA (GMA).
Preferably, described crosslinking agent is selected from any one in ethylene glycol dimethacrylate (EDMA), N, N '-methylene-bisacrylamide (MBAA), polyethyleneglycol diacrylate (PEGDA).
Preferably, described crosslinking agent is N, N '-methylene-bisacrylamide (MBAA).
Preferably, described pore-foaming agent is selected from methyl-sulfoxide, any one or multiple combination in lauryl alcohol, diglycol.
Preferably, described pore-foaming agent is lauryl alcohol and methyl-sulfoxide.
Preferably, described initator is selected from any one in ABVN and azodiisobutyronitrile (AIBN).
Preferably, described initator is azodiisobutyronitrile (AIBN).
Preferably, the weight ratio that described monomer, crosslinking agent, pore-foaming agent and initator add is: 25-35:55-65:360-380:1.
Preferably, the weight ratio that described monomer, crosslinking agent, pore-foaming agent and initator add is: 30:60:374:1.
Preferably, described capillary successively will carry out soda acid activation and γ-MAPS derivatization reaction.
Described soda acid activation refers to described capillary tube inner wall by acid, alkali activation with derivative more silicone hydroxyl.
Described γ-MAPS is derivative refers to that γ-methacrylic acid oxygen propyl trimethoxy silicane (γ-MAPS) reacts with the silicone hydroxyl on capillary tube inner wall with derivative double bond.
Preferably, described pre-polymerization liquid is uniform solution.
Preferably, described mixing condition is: first vortex 4-6 minute, more ultrasonic 25-35 minute.
Preferably, described mixing condition is: first vortex 5 minutes, more ultrasonic 30 minutes.
Preferably, described polymeric reaction condition is: reaction vessel: water-bath; Reaction temperature: 70-80 DEG C; Reaction time: 11-13 hour.
Preferably, described polymeric reaction condition is: reaction vessel: water-bath; Reaction temperature: 75 DEG C; Reaction time: 12 hours.
2) flexible dendrimer is filled into step 1) in obtain have in the base integral post of active group and carry out grafting modification reaction, namely obtain the integral post that flexible dendrimer is modified;
Preferably, described flexible dendrimer, by organic solvent dissolution, injects base integral post.
Preferably, described organic solvent is absolute methanol.
Preferably, described grafting modification reaction condition is: the pressure of pump: 2.9-3.1MPa; Reaction temperature: 55-65 DEG C; Reaction time: 11-13 hour.
Preferably, described grafting modification reaction condition is: the pressure of pump: 3MPa; Reaction temperature: 60 DEG C; Reaction time: 12 hours.
The described flexible dendrimer added can make the monomer complete reaction in post.The concentration of described flexible dendrimer only affects reaction rate.
3) by substituted boracic acid implantation step 2) in carry out modification reaction in the flexible dendrimer that the obtains integral post of modifying, obtain dendroid boric acid functionalization integral post.
Preferably, described substituted boracic acid, by organic solvent dissolution, injects base integral post.
Preferably, described organic solvent is absolute methanol.
Preferably, described modification reaction carries out at twice.
Preferably, modification reaction condition is: reaction temperature: room temperature described first time; Reaction time: 5.5-6.5 hour.
Preferred, described first time modification reaction condition be: reaction temperature: room temperature; Reaction time: 6 hours.
Preferably, in described first time modification reaction, reactant is substituted boracic acid, i.e. 2,4-bis-fluoro-3-formylphenylboronic acids (DFFPBA).
Preferably, described second time modification reaction condition is: reaction temperature: room temperature; Reaction time: 11-13 hour.
Preferred, described second time modification reaction condition is: reaction temperature: room temperature; Reaction time: 12 hours.
Preferably, in described second time modification reaction, reactant is the reducing agent of substituted boracic acid, i.e. sodium cyanoborohydride (NaCNBH
3).
Preferably, in described integral post, unreacted material adopts organic solvent to rinse.Preferably, described organic solvent is absolute methanol.
The described substituted boracic acid that adds and reducing agent thereof can make the primary amine complete reaction on the PEI in post.The concentration of described substituted boracic acid and reducing agent thereof only affects reaction rate.
The present invention further discloses above-mentioned a kind of dendroid boric acid functionalization integral post to the purposes in the separation of the biomolecule containing cis-form dihydroxy structure, enrichment and purifying.
Preferably, described purposes is above-mentioned dendroid boric acid functionalization integral post to the purposes in being separated of glycoprotein and saccharide compound, enrichment and purifying.
Preferably, described purposes be above-mentioned dendroid boric acid functionalization integral post in saliva to the purposes in the separation of glycoprotein, enrichment and purifying.
Preferably, described purposes be above-mentioned dendroid boric acid functionalization integral post in tobacco to the purposes in the separation of saccharide compound, enrichment and purifying.
As mentioned above, the invention discloses a kind of dendroid boric acid functionalization integral post and its preparation method and application, this boric acid functionalization integral post makes boric acid all significantly improve the affinity of biomolecule and binding capacity that are rich in cis-form dihydroxy, has high-affinity, high power capacity.Because flexible dendrimer contains a large amount of surface groups, boric acid aglucon number is amplified after boric acid modified, make material hole surface containing highdensity boric acid base group, multivalence synergy can be produced to the biomolecule being rich in cis-form dihydroxy, thus significantly improve adhesion and binding capacity.Experimental result shows, due to multidigit point synergy, this flexible dendroid boric acid functionalization integral post is 10 to the dissociation constant of glycoprotein
-6~ 10
-7m, is up to 37 μm of ol/mL to micromolecular binding capacity, and compared with the affine organic whole post of boron reported at present, this bond strength and binding capacity value are all the highest.
In addition, it is wide that this integral post also has applicable pH range, and antijamming capability is strong and regulate without the need to pH advantages such as getting final product Direct Analysis to actual sample.Because this post is capillary monolithic column, effective on-line analysis can be carried out to nano-upgrading sample.The flexible dendroid boric acid functionalization integral post of high-affinity of the present invention, high power capacity is very suitable for Selective Separation, the enrichment of the cis-form dihydroxy biomolecule such as low concentration glycoprotein and sugar in complicated reagent sample, as selective extraction and the enrichment of saccharide compound in the glycoprotein in saliva and tobacco sample.
Accompanying drawing explanation
Fig. 1 is shown as the structural representation of flexible dendroid boric acid functionalization integral post
Fig. 2 is shown as flexible dendroid boric acid functionalization integral post scanning electron microscope diagram 2A, 2B, and wherein, 2A is 600 times of enlarged drawings, and 2B is 5000 times of enlarged drawings
Fig. 3 is shown as the IR Characterization figure of flexible dendroid boric acid functionalization integral post, and wherein, a is Epoxy functionalized integral material; B is the integral material that flexible dendrimer is modified; C is flexible dendroid boric acid functionalization integral material
Fig. 4 is shown as flexible dendroid boric acid functionalization integral post that PEI assists reservation Fig. 4 A, 4B, 4C, 4D to adenosine and desoxyadenossine, and wherein 4A is the auxiliary reservation figure of PEI molecular weight 600; 4B is the reservation figure that PEI molecular weight 1800 is assisted; 4C is the reservation figure that PEI molecular weight 10000 is assisted; 4D is the reservation figure that PEI molecular weight 70000 is assisted
Fig. 5 be shown as flexible dendroid boric acid functionalization integral post to adenosine and catechol in conjunction with pH Fig. 5 A, 5B, wherein, 5A is the combination to adenosine; 5B is the combination to catechol
Fig. 6 is shown as flexible dendroid boric acid functionalization integral post to the selective binding figure of glycoprotein
Fig. 7 be shown as flexible dendroid boric acid functionalization integral post to horseradish peroxidase (HRP) and ribonuclease B (RNase B) in conjunction with pH Fig. 7 A, 7B, wherein, 7A is the combination to HRP; 7B is the combination to RNase B
Fig. 8 is shown as the mass spectrogram of glycoprotein in flexible dendroid boric acid functionalization integral post selective enrichment saliva, and wherein, a is the mass spectrogram of saliva Direct Analysis; The mass spectrogram of the flexible dendroid boric acid functionalization integral post of b extract in saliva, peak 1-9 is the glycoprotein identified
Fig. 9 is shown as the electrophoretogram of the sugar solid phase labelling after boric acid functionalization integral post (b) extraction that flexible dendroid boric acid functionalization entirety (a) post and ethylenediamine are assisted in saliva, wherein, a is the extraction of flexible dendroid boric acid functionalization integral post; B is the boric acid functionalization integral post extraction that ethylenediamine is assisted
Detailed description of the invention
Set forth the present invention further below in conjunction with specific embodiment, should be understood that these embodiments are only not used in for illustration of the present invention and limit the scope of the invention.
Below by way of specific instantiation, embodiments of the present invention are described, those skilled in the art the content disclosed by this description can understand other advantages of the present invention and effect easily.The present invention can also be implemented or be applied by detailed description of the invention different in addition, and the every details in this description also can based on different viewpoints and application, carries out various modification or change not deviating under spirit of the present invention.
The reagent used in following examples is as polymine-dendrimer (PEI), 2,4-bis-fluoro-3-formylphenylboronic acids (DFFPBA), sodium cyanoborohydride (NaCNBH
3), γ-methacrylic acid oxygen propyl trimethoxy silicane (γ-MAPS), ethylene glycol dimethacrylate (EDMA), GMA (GMA), N, N '-methylene-bisacrylamide (MBAA), lauryl alcohol, methyl-sulfoxide, diglycol, ABVN, azodiisobutyronitrile (AIBN), absolute methanol (>=99.5%), desoxyadenossine, adenosine, sodium dihydrogen phosphate/buffer solution of sodium phosphate acetic acid, catechol, horseradish peroxidase (HRP), glycoprotein, transferrins (TRF), ribonuclease A (RNase A), ribonuclease B (RNaseB), protease inhibitors, ammonium acetate buffer solution, acetonitrile (ACN), sodium chloride (NaCl), tobacco leaf, 1-phenyl-3-methyl-5-pyrazolones ketone (PMP), borax (>=99.5%) all can be commercially.
The equipment used in following examples as capillary, turbula shaker, ultrasonic extraction instrument, water-bath, self-control pump, HPLC column, apparatus,Soxhlet's, Fourier transform-infrared spectrometer (FT-IR), elemental analyser, high performance liquid chromatograph, ultra-violet and visible spectrophotometer, the boric acid functionalization integral post that ethylenediamine is auxiliary, HPCE, 4800plus type MALDI-TOF MS (AB Sciex mass spectrometer system company), is routine and can equipment commercially.
Embodiment 1: the preparation of Epoxy functionalized integral post 1
Select integral post based on Epoxy functionalized integral post.Epoxy functionalized integral post adopts heat to cause Raolical polymerizable synthesis.Before preparation integral post, the derivative process of acid, alkali activation and γ-MAPS is successively carried out to capillary, afterwards first by monomer methacrylic acid ethylene oxidic ester (GMA) 30mg, the crosslinking agent N containing epoxy functionality, N '-methylene-bisacrylamide (MBAA) 60mg, pore-foaming agent lauryl alcohol 174mg and methyl-sulfoxide 200mg altogether 374mg mix with initator azodiisobutyronitrile (AIBN) 1mg, vortex 5 minutes, then ultrasonic 30 minutes, homogeneous pre-polymerization liquid is obtained.Be filled in the capillary after double bond is modified, then put into 75 DEG C of water-bath reactions 12 hours.After polymerisation completes, with methyl alcohol unreacted material flushed out and namely obtain Epoxy functionalized integral post.
Embodiment 2: the preparation of Epoxy functionalized integral post 2
Select integral post based on Epoxy functionalized integral post.Epoxy functionalized integral post adopts heat to cause Raolical polymerizable synthesis.Before preparation integral post, the derivative process of acid, alkali activation and γ-MAPS is successively carried out to capillary, afterwards first by monomer methacrylic acid ethylene oxidic ester (GMA) 25mg, the crosslinking agent N containing epoxy functionality, N '-methylene-bisacrylamide (MBAA) 55mg, pore-foaming agent lauryl alcohol 160mg and methyl-sulfoxide 200mg altogether 360mg mix with initator azodiisobutyronitrile (AIBN) 1mg, vortex 4 minutes, then ultrasonic 25 minutes, homogeneous pre-polymerization liquid is obtained.Be filled in the capillary after double bond is modified, then put into 70 DEG C of water-bath reactions 11 hours.After polymerisation completes, with methyl alcohol unreacted material flushed out and namely obtain Epoxy functionalized integral post.
Embodiment 3: the preparation of Epoxy functionalized integral post 3
Select integral post based on Epoxy functionalized integral post.Epoxy functionalized integral post adopts heat to cause Raolical polymerizable synthesis.Before preparation integral post, the derivative process of acid, alkali activation and γ-MAPS is successively carried out to capillary, afterwards first by monomer methacrylic acid ethylene oxidic ester (GMA) 35mg, the crosslinking agent N containing epoxy functionality, N '-methylene-bisacrylamide (MBAA) 65mg, pore-foaming agent lauryl alcohol 180mg and methyl-sulfoxide 200mg altogether 380mg mix with initator azodiisobutyronitrile (AIBN) 1mg, vortex 6 minutes, then ultrasonic 35 minutes, homogeneous pre-polymerization liquid is obtained.Be filled in the capillary after double bond is modified, then put into 80 DEG C of water-bath reactions 13 hours.After polymerisation completes, with methyl alcohol unreacted material flushed out and namely obtain Epoxy functionalized integral post.
Embodiment 4: the preparation of flexible polyethylene imines (PEI) dendrimer functionalization integral post
Select polymine-dendrimer (PEI) as flexible dendrimer.Embodiment 1 is obtained containing Epoxy functionalized base integral post respectively with polymine (PEI) the compound generation epoxy ring-opening reaction of different molecular weight (600,1800,10000 and 70000).Under the pressure of pump, be that the PEI methanol solution of 100mg/mL to inject in post 60 DEG C of reactions 12 hours by concentration.With methyl alcohol unreacted material flushed out after modification reaction completes and namely obtain flexible dendrimer functionalization integral post.
Embodiment 5: the preparation of flexible dendroid boric acid functionalization integral post
The dendrimer functionalization integral post of the various different molecular weights that embodiment 4 is obtained successively respectively with DFFPBA/MeOH (50mg/mL) and NaBH
3cN/MeOH (100mg/mL) reacts 6 hours and 12 hours at ambient temperature, with methyl alcohol, unreacted material is flushed out the flexible dendroid boric acid functionalization integral post namely obtained as shown in Figure 1 after modification reaction completes.As shown in Figure 2, Fig. 2 A illustrates that this integral post is connected with post jamb well to the morphology characterization of this integral post, and Fig. 2 B illustrates and presents good skeleton structure and through hole.
Embodiment 6: the preparation of large volume flexible dendroid boric acid functionalization integral material
According to as when preparing integral post in embodiment 1 the reagent proportioning that adopts, i.e. monomer GMA 30mg, crosslinking agent MBAA60mg, pore-foaming agent lauryl alcohol 174mg, methyl-sulfoxide 200mg 374mg and initiator A IBN 1mg altogether, having prepared pre-polymerization liquid inserts inside conventional HPLC column, then at the temperature of 75 DEG C after each reaction 12h, go out the base integral material of generation, and thinly slice.Then, thin slice is used methanol extraction 24h in apparatus,Soxhlet's, under 60 DEG C of vacuum, namely dry 24h has prepared large volume Epoxy functionalized integral material name M-A subsequently.Then the PEI methanol solution of this thin slice of part and various molecular weight (600,1800,10000 and 70000) is mixed, stirring reaction 12h at 60 DEG C.At apparatus,Soxhlet's methanol wash 24h after filtration, dry 12h under 60 DEG C of vacuum subsequently, the flexible dendroid functionalization integral material (naming M-B1 respectively, M-B2, M-B3 and M-B4) obtained thus respectively with containing DFFPBA, NaBH
3the solution of CN, ACN and MeOH mixes, and reacts 12h at normal temperatures.After filtration, in apparatus,Soxhlet's, use methanol wash 24h, subsequently dry 12h under 60 DEG C of vacuum.The large volume obtained thus flexible dendroid boric acid functionalization integral material (names M-C1 respectively, M-C2, M-C3 and M-C4) to characterize as Fourier transform-infrared spectrum (FT-IR) and elementary analysis to confirm that each walks being successfully completed of modification reaction.Each step In situ FTIR sign and elementary analysis are respectively as shown in Figure 3 and Table 1.
As shown in Figure 3, the epoxy-functional on integral material M-A is 995 and 753cm
-1between have absworption peak, after PEI open loop and further boric acid functionalization, these absworption peaks disappear, and illustrate that epoxide group and amino-compound successfully there occurs nucleophilic ring opening addition reaction.
Table 1 results of elemental analyses
| Integral material | C(％) | H(％) | N(％) |
| M-A | 49.71 | 6.74 | 10.61 |
| M-B1 | 50.21 | 7.42 | 12.26 |
| M-B2 | 49.74 | 7.52 | 12.40 |
| M-B3 | 49.81 | 7.46 | 12.60 |
| M-B4 | 50.11 | 7.45 | 11.98 |
| M-C1 | 50.83 | 7.51 | 11.41 |
| M-C2 | 50.86 | 7.49 | 11.39 |
| M-C3 | 50.91 | 7.52 | 11.25 |
| M-C4 | 51.88 | 7.71 | 11.31 |
As shown in table 1, the upper N element percentage composition of M-B, apparently higher than N element percentage composition on M-A, illustrates that the epoxy-functional on M-A and the primary amine on PEI there occurs nucleophilic ring opening reaction.In addition, the N element percentage composition on M-C is starkly lower than the N element percentage composition on M-B, the C on M-C, and H element percentage composition is apparently higher than the C on M-B, and H element percentage composition, illustrates that boric acid base group is successfully modified on integral material by schiff base reaction.
Embodiment 7: the investigation of the affine performance of flexible dendroid boric acid functionalization integral post boron
Investigate as analyzing thing the affine performance of the flexible boron of dendroid boric acid functionalization integral post under neutral environment that embodiment 5 obtains with desoxyadenossine and adenosine.Adenosine is a material containing cis-form dihydroxy structure, and desoxyadenossine is not then containing cis-form dihydroxy structure, so desoxyadenossine can the label in thing and dead time in contrast.
Adopt high performance liquid chromatography can carry out adenosine combination and release experiment to flexible dendroid boric acid functionalization integral post.The basic parameter of instrument arranges as follows: sample-loading buffer: 50mM phosphate (pH=7.0); Eluent: 100mM acetum; Sample: 0.5mg/mL adenosine (A) and desoxyadenossine (DA) solution; Flow velocity: 1.0 μ L/min; Sample size: 100nL; Determined wavelength: 254nm.
Experimentation is summarized as follows: first use buffer solution (50mM phosphate, pH=7.0) to balance pillar, then sample introduction; Then use wash buffer 25 minutes, then switch to the sample that 100mM acetum wash-out is caught by pillar.As shown in Figure 4, because boric acid can become ring esterification with cis-form dihydroxy, so the flexible dendroid boric acid functionalization integral post containing different molecular weight PEI can capture adenosine and not capture desoxyadenossine.Result shows that the flexible dendroid boric acid functionalization integral post that embodiment 5 obtains can carry out specific recognition to the biomolecule containing cis-form dihydroxy structure.
Embodiment 8: the investigation of flexible dendroid boric acid functionalization integral post binding capacity
Reserve capacity is the important indicator evaluating boron affine integral post, and the concentrated expression affine integral post of boron is to the binding ability of the dihydroxylation compound of cis.Take adenosine as test substances, the reserve capacity of flexible dendroid boric acid functionalization integral post under different pH condition obtained by the embodiment 3 that frontier chromatography determines four kinds of different PEI molecular weight.Wherein desoxyadenossine can not be retained by the affine integral post of boron and flow out in the dead time, can as the label in dead time.Described frontier chromatography and frontal chromatography, sample itself is done mobile phase continually by chromatographic column, adsorb or dissolve the most weak component, first chromatographic column is gone out with pure material state flow, what then flow out in turn is the mixture that time weak component and first flows out component, the like, thus realize the chromatography of mixture separation.
First flexible dendroid boric acid functionalization integral post activates 30min with 100mM HAc, then uses loading buffer (having the 50mM PBS of different pH value) to balance 30min.Again the sample-loading buffer containing 0.5mg/mL adenosine and 0.05mg/mL desoxyadenossine mixture is passed through integral post with the flow velocity of 1.0 μ L/min.After pillar is saturated by adenosine, saturated platform will be there will be, then with the adenosine that 100mM HAc wash-out is retained, then balance integral post with corresponding sample-loading buffer.Circulate with this, under mensuration condition of different pH, the reserve capacity of integral post, the results are shown in table 2.Result shows that the binding capacity of the flexible dendroid boric acid functionalization integral post that the PEI of molecular weight 10000 assists is the highest, also be simultaneously that in current similar integral post, capacity is the highest, therefore all come by this flexible dendroid boric acid functionalization integral post in conjunction with the enrichment of glycoprotein in the investigation of the selective and glycoprotein affinity of pH, glycoprotein and saliva.
The binding capacity of table 2 flexible dendroid boric acid functionalization integral post
Embodiment 9: flexible dendroid boric acid functionalization integral post is in conjunction with the investigation of pH
Flexible dendroid boric acid functionalization integral post embodiment 5 obtained, under having investigated condition of different pH according to the method in embodiment 7, flexible dendroid boric acid functionalization integral post is to the reservation of adenosine and catechol, and concrete outcome is shown in Fig. 5.Wherein, the basic parameter of instrument arranges as follows: sample-loading buffer: 50mM phosphate (pH=5.0-7.0); Eluent: 100mM acetum; Sample: 0.5mg/mL adenosine or catechol solution; Sample size: 100nL; Flow velocity: 1.0 μ L/min; Determined wavelength: 254nm.
As shown in Figure 5, because the pK of DFFPBA
abe worth lower, and the pliability of PEI is stronger, so when mobile phase pH from 7.0 to 5.5 time, this flexible dendroid boric acid functionalization integral post still has adenosine and catechol and retains completely, and namely this integral post can work under neutral and weakly acidic physiological condition.
Embodiment 10: flexible dendroid boric acid functionalization integral post is to the selective of glycoprotein with in conjunction with pH
The flexible dendroid boric acid functionalization integral post adopting high performance liquid chromatography embodiment 5 to be obtained carries out combination and the release experiment of glycoprotein.The basic parameter of instrument arranges as follows: sample-loading buffer: 50mM phosphate (pH=5.5-7.5); Eluent: 100mM acetum; Flow velocity: 1.0 μ L/min; Sample size: 800nL; Determined wavelength: 214nm; Sample 1:1mg/mL horseradish peroxidase (HRP), ribonuclease B (RNase B) and ribonuclease A (RNase A) solution; Sample 2:1mg/mL glycoprotein solution.
Experimentation is summarized as follows: first use sample solution (50mM phosphate, pH=7.5,7.0,6.5,6.0,5.5) to balance pillar, then sample introduction, the glycoprotein h RP in sample and RNase B is caught by the boric acid base group of hole surface through integral post; Then rinse 25 minutes with sample solution, then switch to 100mM acetum, the glycoprotein that wash-out is caught by pillar, experimental result as shown in Figures 6 and 7.As Fig. 6 and 7 shows, this material is because of can not only selective crawl glycoprotein containing boric acid base group, and to containing the HRP of multiple glycosylation site minimum be 6.5 in conjunction with pH, far below similar Monolithic Columns, this illustrates that the boric acid aglucon number amplified through PEI can to the macromolecular compound containing multiple cis-form dihydroxy as glycoprotein produces multivalence synergy, thus substantially increase adhesion, reduce in conjunction with pH, therefore can be applied directly to most of actual sample and regulate without the need to pH.
Embodiment 11: flexible dendroid boric acid functionalization integral post is to the investigation of glycoprotein affinity
The glycoprotein standard liquid of a series of concentration is prepared with 50mM PBS (pH=7.4).Under nitrogen pressure, 60 μ L phosphate buffer (50mM, pH=7.4) are first used to balance the flexible dendroid boric acid functionalization integral post of certain length; Pass through this integral post with 110 or 400 μ L glycoprotein solution again, and collect efflux; Then this integral post is washed with 10 μ L phosphate buffers; Finally with the protein that 110 μ L 100mM acetum wash-outs are combined by integral post, and collect eluent.Carry out ultraviolet absorptivity detection to serial glycoprotein standard liquid, collection solution and elute soln, HRP determined wavelength is 403nm, and transferrins (TRF) and RNase B determined wavelength are 280nm.Dissociation constant (K is calculated according to Scatchard equation
d) and apparent maximum binding capacity (Q
max):
Q in formula
maxfor the saturated adsorption capacity of flexible dendroid boric acid functionalization integral post, Q
efor flexible dendroid boric acid functionalization integral post is to the equilibrium adsorption capacity of glycoprotein, C
sfor the free glycoprotein concentration when adsorption equilibrium, K
dfor dissociation constant.Q
e/ C
sto Q
ecarry out linear regression, its slope is-1/K
d, intercept is Q
max/ K
d, can dissociation constant K be calculated
dwith saturated adsorption capacity Q
max.Measurement result is as shown in table 3.
Result shows, due to multidigit point synergy, this flexible dendroid boric acid functionalization integral post is 10 to the dissociation constant of glycoprotein
-6~ 10
-7m, meanwhile, glycosylation site number is more, and multivalence synergy is higher, and adhesion is stronger.In addition, this material is suitable to the saturated extent of adsorption of HRP and RNase B, and description taken in conjunction power does not affect saturated extent of adsorption, and the saturated extent of adsorption of transferrins is lower is because the molecular volume of this albumen hinders more greatly the combination in boric acid site.
The dissociation constant of table 3 flexible dendroid boric acid functionalization integral post and glycoprotein and saturated adsorption capacity
| Glycoprotein | Dissociation constant | Saturated extent of adsorption (μm ol/g) | Glycosylation site |
| Horseradish peroxidase | (2.81±0.28)×10 -7 | 0.32±0.03 | 9 |
| Ribonuclease B | (9.52±1.01)×10 -6 | 0.26±0.01 | 1 |
| Transferrins | (4.44±0.57)×10 -7 | 0.11±0.01 | 2 |
Embodiment 12: the glycoprotein in flexible dendroid boric acid functionalization integral post selective enrichment saliva
Saliva sample prepares: morning is from the saliva of last dietary intake at least 2 h before harvest Healthy People.Before collection, first gargle 3 times with water.When collecting saliva, the test tube holding saliva is placed on ice.In order to reduce the degraded of protein, after saliva collection, add protease inhibitors (1mL saliva adds 1 μ L protease inhibitors) immediately.Then at 4 DEG C, with the centrifugation 10 minutes of 12000 rpms.Collect supernatant liquor and be stored in-80 DEG C.Before use, at 4 DEG C, redissolve sample, can directly use without the need to adjust ph.
First use 50mM, it is the flexible dendroid boric acid functionalization integral post that the embodiment 5 of 250 μm obtains that pH=7.0 phosphate buffer balances long 30cm internal diameter; This integral post is passed through again with the saliva sample that 200 μ L collect; Then pillar is rinsed with 100 μ L containing the ammonium acetate buffer solution (pH=7.0) of the 0.25M of 0.50M NaCl and 1M urea and 50 μ L 0.02M ammonium acetate buffer solution (pH=7.0) respectively, finally with the protein that 100mM acetum wash-out flexible dendroid boric acid functionalization integral post is caught, every 5 μ L collect a pipe and collect 10 pipes altogether, and eluent uses MALDI-TOF MS to analyze.
Analysis result as shown in Figure 8, because the concentration of glycoprotein is very low and there is the signal disturbing of abundant non-glycoprotein and carbohydrate in saliva sample, so directly carry out mass spectral analysis to saliva can not glycoprotein be detected, and the saliva after flexible dendroid boric acid functionalization integral post extraction is because being enriched glycoprotein and eliminating other interfering component, therefore multiple glycoprotein can be detected by mass spectral analysis.Result shows the glycoprotein of trace in flexible dendroid boric acid functionalization integral post energy enrichment complex system.
Embodiment 13: flexible dendroid boric acid functionalization integral post is to the selective extraction of saccharide compound in tobacco sample and be separated detection
From cigarette, obtain about 500mg tobacco leaf, add 5ml 50% methanol aqueous solution, stir ultrasonic extraction 30min at 70 DEG C, centrifugation obtains supernatant 2mL, adds 2ml pH=8.0 PBS (PBS), obtains sample solution 4mL.
Get above-mentioned tobacco sample 1mL respectively and pass into the auxiliary boric acid functionalization integral post of flexible dendroid boric acid functionalization integral post and ethylenediamine with the flow velocity of 1.5 μ L/min, then pillar is rinsed with 100 μ L PBSs, pass into 1-phenyl-3-methyl-5-pyrazolones ketone (PMP) solution (preparation of PMP solution: get 17.4mgPMP and add 800 μ L methyl alcohol and 200 μ L pH=8.0 PBSs and dissolve obtained) of 50 μ L 0.1mol/L afterwards at a certain temperature, then 100 μ L washed with methanol pillars are used, the sample solution that the subsequent analysis that solution that desorb obtains is us of carrying out the acetum finally passing into 100 μ L0.1mol/L detects.
Get above-mentioned sample solution 10 μ L and carry out electrophoretic analysis, deposition condition is as follows: capillary overall length: 66cm; Internal diameter: 75 μm; The borax soln of cushioning liquid: 50mmol/L; PH:10.5; Adopt hydrodynamic injection, pressure: 0.5psi; Sample injection time: 15s; Temperature: 25 DEG C.
Analysis result as shown in Figure 9, because saccharide compound does not have UV absorption, utilizes electrophoresis cannot detect saccharide compound in tobacco by Direct Analysis, therefore need first with labelled reagent as PMP mark after sugar in tobacco just can be detected.If but directly carry out traditional liquid phase mark extracting in the sugar juice after tobacco, PMP can cause interference to the signal of sugar, carries out solid phase labelling again and can avoid this unfavorable factor after the sugar now utilizing the affine integral post of boron to catch in tobacco.The boric acid functionalization integral post auxiliary relative to traditional ethylenediamine, the flexible dendroid boric acid functionalization integral post that binding capacity high-bond is strong extracts tobacco and component after solid phase labelling can detect more strong peak by electrophoretic analysis.Result shows that flexible dendroid boric acid functionalization integral post can saccharide compound in more effective enrichment tobacco.
Above-described embodiment is illustrative principle of the present invention and effect thereof only, but not for limiting the present invention.Any person skilled in the art scholar all without prejudice under spirit of the present invention and category, can modify above-described embodiment or changes.Therefore, such as have in art usually know the knowledgeable do not depart from complete under disclosed spirit and technological thought all equivalence modify or change, must be contained by claim of the present invention.
Claims (10)
1. a dendroid boric acid functionalization integral post, is characterized in that, it is the flexible dendroid boric acid functionalization integral post obtained through substituted boracic acid functionalization after the flexible dendrimer of base integral post holes surface bond.
2. a kind of dendroid boric acid functionalization integral post according to claim 1, it is characterized in that, described base integral post is Organic Polymer Monolithic Columns; Described Organic Polymer Monolithic Columns be selected from Epoxy functionalized Organic Polymer Monolithic Columns or other contain in the Organic Polymer Monolithic Columns of active group any one.
3. a kind of dendroid boric acid functionalization integral post according to claim 1, is characterized in that, described flexible dendrimer be selected from polymine-dendrimer or other dendrimer any one; Described substituted boracic acid be selected from formylphenylboronic acid or Carboxybenzeneboronic acid any one.
4. a preparation method for dendroid boric acid functionalization integral post, specifically comprises the following steps:
1) obtain pre-polymerization liquid after the monomer containing active group, crosslinking agent, pore-foaming agent and initator being mixed, be filled in capillary, carry out polymerisation, synthesis obtains the base integral post with active group;
2) flexible dendrimer is filled into step 1) in obtain have in the base integral post of active group and carry out grafting modification reaction, namely obtain the integral post that flexible dendrimer is modified;
3) by substituted boracic acid implantation step 2) in carry out modification reaction in the flexible dendrimer that the obtains integral post of modifying, obtain dendroid boric acid functionalization integral post.
5. the preparation method of a kind of dendroid boric acid functionalization integral post according to claim 4, it is characterized in that, step 1) in, the described monomer containing active group is selected from containing epoxy functionality, carboxyl functional group, mercapto functional group, amido functional group, any one in the monomer of hydroxy functional group and aldehyde functions; Described crosslinking agent be selected from ethylene glycol dimethacrylate, N, N '-methylene-bisacrylamide, polyethyleneglycol diacrylate any one; Described pore-foaming agent is selected from methyl-sulfoxide, any one or multiple combination in lauryl alcohol, diglycol; Described initator be selected from ABVN and azodiisobutyronitrile any one.
6. the preparation method of a kind of dendroid boric acid functionalization integral post according to claim 4, is characterized in that, step 1) in, described mixing condition is: first vortex 4-6 minute, more ultrasonic 25-35 minute; Described polymeric reaction condition is: reaction vessel: water-bath, reaction temperature: 70-80 DEG C, the reaction time: 11-13 hour.
7. the preparation method of a kind of dendroid boric acid functionalization integral post according to claim 4, is characterized in that, step 2) in, described grafting modification reaction condition is: the pressure of pump: 2.9-3.1MPa; Reaction temperature: 55-65 DEG C; Reaction time: 11-13 hour.
8. the preparation method of a kind of dendroid boric acid functionalization integral post according to claim 4, is characterized in that, step 3) in, described modification reaction carries out at twice; Described first time, modification reaction condition was: reaction temperature: room temperature, reaction time: 5.5-6.5 hour; Described second time modification reaction condition is: reaction temperature: room temperature, reaction time: 11-13 hour.
9. according to described a kind of dendroid boric acid functionalization integral post arbitrary in claim 1-3 to the purposes in the separation of the biomolecule containing cis-form dihydroxy structure, enrichment and purifying.
10. the purposes of a kind of dendroid boric acid functionalization integral post according to claim 9, is characterized in that, described purposes is described dendroid boric acid functionalization integral post to the purposes in being separated of glycoprotein and saccharide compound, enrichment and purifying.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108927216A (en) * | 2018-06-29 | 2018-12-04 | 同济大学 | A kind of porous carrier catalysis material and the preparation method and application thereof of patch constraint |
| CN109206614A (en) * | 2017-07-05 | 2019-01-15 | 大连医科大学 | A kind of reagent and its preparation method and application of the detection containing sugar chain biomolecule |
| CN110314673A (en) * | 2019-08-08 | 2019-10-11 | 福州大学 | A kind of affine integral post of aptamer functionalization and preparation method thereof based on light-initiated hybrid polymer |
| CN110698697A (en) * | 2019-08-30 | 2020-01-17 | 厦门大学 | Preparation method of polyethyleneimine-polyvinyl alcohol hydrogel with self-healing performance |
| CN111139052A (en) * | 2018-11-06 | 2020-05-12 | 中国石油化工股份有限公司 | Low-concentration hydroxypropyl guar gum fracturing fluid and preparation method thereof |
| CN111139053A (en) * | 2018-11-06 | 2020-05-12 | 中国石油化工股份有限公司 | Nano silicon dioxide organic boron crosslinking agent and preparation method thereof |
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| CN111729661A (en) * | 2020-06-09 | 2020-10-02 | 苏州赛分科技有限公司 | Chromatography media for separating boron-containing species |
| CN112337441A (en) * | 2020-09-23 | 2021-02-09 | 江苏理工学院 | Phenylboronic acid-chitosan modified monolithic column and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102059103A (en) * | 2010-12-08 | 2011-05-18 | 南京大学 | Hydrophylic phenylboric acid functional porous integral material, preparation method and application thereof |
| CN103157447A (en) * | 2013-03-22 | 2013-06-19 | 南京大学 | High-affinity tree-like boric acid functionalized magnetic microsphere as well as preparation method and application thereof |
| CN103550954A (en) * | 2013-11-14 | 2014-02-05 | 福州大学 | Phenylimidazole ionic liquid-silica gel hybrid capillary tube monolithic column |
| WO2014184810A1 (en) * | 2013-05-15 | 2014-11-20 | Council Of Scientific & Industrial Research | Method for assaying glycated protein |
-
2014
- 2014-12-29 CN CN201410856918.6A patent/CN104549182B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102059103A (en) * | 2010-12-08 | 2011-05-18 | 南京大学 | Hydrophylic phenylboric acid functional porous integral material, preparation method and application thereof |
| CN103157447A (en) * | 2013-03-22 | 2013-06-19 | 南京大学 | High-affinity tree-like boric acid functionalized magnetic microsphere as well as preparation method and application thereof |
| WO2014184810A1 (en) * | 2013-05-15 | 2014-11-20 | Council Of Scientific & Industrial Research | Method for assaying glycated protein |
| CN103550954A (en) * | 2013-11-14 | 2014-02-05 | 福州大学 | Phenylimidazole ionic liquid-silica gel hybrid capillary tube monolithic column |
Non-Patent Citations (1)
| Title |
|---|
| 刘云春: "苯硼酸功能化整体柱的制备、应用及非常规作用机理研究", 《中国博士学位论文全文数据库 工程科技1辑》 * |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109206614B (en) * | 2017-07-05 | 2020-10-16 | 大连医科大学 | Reagent for detecting sugar chain-containing biomolecule and preparation method and application thereof |
| CN109206614A (en) * | 2017-07-05 | 2019-01-15 | 大连医科大学 | A kind of reagent and its preparation method and application of the detection containing sugar chain biomolecule |
| CN108927216A (en) * | 2018-06-29 | 2018-12-04 | 同济大学 | A kind of porous carrier catalysis material and the preparation method and application thereof of patch constraint |
| CN111139052B (en) * | 2018-11-06 | 2022-06-10 | 中国石油化工股份有限公司 | Low-concentration hydroxypropyl guanidine gum fracturing fluid and preparation method thereof |
| CN111139053B (en) * | 2018-11-06 | 2022-05-13 | 中国石油化工股份有限公司 | Nano silicon dioxide organic boron crosslinking agent and preparation method thereof |
| CN111139052A (en) * | 2018-11-06 | 2020-05-12 | 中国石油化工股份有限公司 | Low-concentration hydroxypropyl guar gum fracturing fluid and preparation method thereof |
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| CN110314673B (en) * | 2019-08-08 | 2021-06-01 | 福州大学 | Aptamer functionalized affinity monolithic column based on photo-initiated hybrid polymerization and preparation method thereof |
| CN110314673A (en) * | 2019-08-08 | 2019-10-11 | 福州大学 | A kind of affine integral post of aptamer functionalization and preparation method thereof based on light-initiated hybrid polymer |
| WO2021037269A1 (en) * | 2019-08-30 | 2021-03-04 | 厦门大学 | Method for preparing polyethyleneimine-polyvinyl alcohol hydrogel having self-healing properties |
| CN110698697A (en) * | 2019-08-30 | 2020-01-17 | 厦门大学 | Preparation method of polyethyleneimine-polyvinyl alcohol hydrogel with self-healing performance |
| CN111272911A (en) * | 2020-03-19 | 2020-06-12 | 中国烟草总公司郑州烟草研究院 | Sample pretreatment method for determining Amadori compound in food |
| CN111579467A (en) * | 2020-06-01 | 2020-08-25 | 天津医科大学 | Bifunctional graphene oxide composite material and application thereof in detecting adherent cells |
| CN111579467B (en) * | 2020-06-01 | 2023-03-14 | 天津医科大学 | Bifunctional graphene oxide composite material and application thereof in detecting adherent cells |
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