CN106111131A - A kind of dendroid plation nano-particle analogue enztme and its preparation method and application - Google Patents
A kind of dendroid plation nano-particle analogue enztme and its preparation method and application Download PDFInfo
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- CN106111131A CN106111131A CN201610484771.1A CN201610484771A CN106111131A CN 106111131 A CN106111131 A CN 106111131A CN 201610484771 A CN201610484771 A CN 201610484771A CN 106111131 A CN106111131 A CN 106111131A
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- 239000002105 nanoparticle Substances 0.000 title claims abstract description 94
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 241000555268 Dendroides Species 0.000 title claims abstract description 19
- 102000004190 Enzymes Human genes 0.000 claims abstract description 25
- 108090000790 Enzymes Proteins 0.000 claims abstract description 25
- 229910045601 alloy Inorganic materials 0.000 claims abstract description 18
- 239000000956 alloy Substances 0.000 claims abstract description 18
- 239000000203 mixture Substances 0.000 claims abstract description 9
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Substances [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 62
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 42
- 239000010931 gold Substances 0.000 claims description 41
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 30
- 238000006243 chemical reaction Methods 0.000 claims description 26
- 235000010323 ascorbic acid Nutrition 0.000 claims description 19
- 239000011668 ascorbic acid Substances 0.000 claims description 19
- 229960005070 ascorbic acid Drugs 0.000 claims description 19
- 239000000463 material Substances 0.000 claims description 15
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 14
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 14
- 239000011591 potassium Substances 0.000 claims description 14
- 229910052700 potassium Inorganic materials 0.000 claims description 14
- 229910052717 sulfur Inorganic materials 0.000 claims description 14
- 239000011593 sulfur Substances 0.000 claims description 14
- YZCKVEUIGOORGS-IGMARMGPSA-N Protium Chemical compound [1H] YZCKVEUIGOORGS-IGMARMGPSA-N 0.000 claims description 13
- 238000001514 detection method Methods 0.000 claims description 12
- 229910052697 platinum Inorganic materials 0.000 claims description 10
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 8
- 229910052737 gold Inorganic materials 0.000 claims description 8
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 4
- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 0.000 claims description 4
- 150000002978 peroxides Chemical class 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims 2
- 229930003268 Vitamin C Natural products 0.000 claims 2
- 235000019154 vitamin C Nutrition 0.000 claims 2
- 239000011718 vitamin C Substances 0.000 claims 2
- 238000002242 deionisation method Methods 0.000 claims 1
- 238000007254 oxidation reaction Methods 0.000 abstract description 24
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract description 21
- 102000003992 Peroxidases Human genes 0.000 abstract description 21
- 108040007629 peroxidase activity proteins Proteins 0.000 abstract description 21
- 230000003647 oxidation Effects 0.000 abstract description 20
- 238000006555 catalytic reaction Methods 0.000 abstract description 17
- 230000000694 effects Effects 0.000 abstract description 15
- 230000003197 catalytic effect Effects 0.000 abstract description 12
- 239000000758 substrate Substances 0.000 abstract description 5
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical class CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 57
- 239000007864 aqueous solution Substances 0.000 description 23
- 239000008367 deionised water Substances 0.000 description 19
- 229910021641 deionized water Inorganic materials 0.000 description 19
- 102000004316 Oxidoreductases Human genes 0.000 description 13
- 108090000854 Oxidoreductases Proteins 0.000 description 13
- SJUCACGNNJFHLB-UHFFFAOYSA-N O=C1N[ClH](=O)NC2=C1NC(=O)N2 Chemical compound O=C1N[ClH](=O)NC2=C1NC(=O)N2 SJUCACGNNJFHLB-UHFFFAOYSA-N 0.000 description 10
- 238000002835 absorbance Methods 0.000 description 9
- 238000010521 absorption reaction Methods 0.000 description 8
- 230000031709 bromination Effects 0.000 description 7
- 238000005893 bromination reaction Methods 0.000 description 7
- 230000003278 mimic effect Effects 0.000 description 7
- 238000000034 method Methods 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- 238000000862 absorption spectrum Methods 0.000 description 5
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 5
- 238000004847 absorption spectroscopy Methods 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- JUWSSMXCCAMYGX-UHFFFAOYSA-N gold platinum Chemical compound [Pt].[Au] JUWSSMXCCAMYGX-UHFFFAOYSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
- 238000005275 alloying Methods 0.000 description 3
- 238000003018 immunoassay Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000004088 simulation Methods 0.000 description 3
- HYHCSLBZRBJJCH-UHFFFAOYSA-M sodium hydrosulfide Chemical compound [Na+].[SH-] HYHCSLBZRBJJCH-UHFFFAOYSA-M 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 125000000217 alkyl group Chemical group 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 238000003759 clinical diagnosis Methods 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- GDSOZVZXVXTJMI-SNAWJCMRSA-N (e)-1-methylbut-1-ene-1,2,4-tricarboxylic acid Chemical compound OC(=O)C(/C)=C(C(O)=O)\CCC(O)=O GDSOZVZXVXTJMI-SNAWJCMRSA-N 0.000 description 1
- ALLIZEAXNXSFGD-UHFFFAOYSA-N 1-methyl-2-phenylbenzene Chemical group CC1=CC=CC=C1C1=CC=CC=C1 ALLIZEAXNXSFGD-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 230000010718 Oxidation Activity Effects 0.000 description 1
- 229910001260 Pt alloy Inorganic materials 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- QYSYEILYXGRUOM-UHFFFAOYSA-N [Cl].[Pt] Chemical compound [Cl].[Pt] QYSYEILYXGRUOM-UHFFFAOYSA-N 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910000420 cerium oxide Inorganic materials 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229910000428 cobalt oxide Inorganic materials 0.000 description 1
- IVMYJDGYRUAWML-UHFFFAOYSA-N cobalt(ii) oxide Chemical compound [Co]=O IVMYJDGYRUAWML-UHFFFAOYSA-N 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- OMZSGWSJDCOLKM-UHFFFAOYSA-N copper(II) sulfide Chemical compound [S-2].[Cu+2] OMZSGWSJDCOLKM-UHFFFAOYSA-N 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229910021389 graphene Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- GPRLSGONYQIRFK-UHFFFAOYSA-N hydron Chemical compound [H+] GPRLSGONYQIRFK-UHFFFAOYSA-N 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 229910000510 noble metal Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- BMMGVYCKOGBVEV-UHFFFAOYSA-N oxo(oxoceriooxy)cerium Chemical compound [Ce]=O.O=[Ce]=O BMMGVYCKOGBVEV-UHFFFAOYSA-N 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Classifications
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- 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
- B01J23/00—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00
- B01J23/38—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00 of noble metals
- B01J23/48—Silver or gold
- B01J23/52—Gold
-
- 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
- B01J35/00—Catalysts, in general, characterised by their form or physical properties
- B01J35/40—Catalysts, in general, characterised by their form or physical properties characterised by dimensions, e.g. grain size
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01J35/00—Catalysts, in general, characterised by their form or physical properties
- B01J35/60—Catalysts, in general, characterised by their form or physical properties characterised by their surface properties or porosity
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F1/00—Metallic powder; Treatment of metallic powder, e.g. to facilitate working or to improve properties
- B22F1/07—Metallic powder characterised by particles having a nanoscale microstructure
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F9/00—Making metallic powder or suspensions thereof
- B22F9/16—Making metallic powder or suspensions thereof using chemical processes
- B22F9/18—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds
- B22F9/24—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds starting from liquid metal compounds, e.g. solutions
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- 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
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
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- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
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Abstract
The invention discloses a kind of plation nano-particle analogue enztme and its preparation method and application.Dendroid plation nano-particle analogue enztme, its plation nano-particle pattern is dendroid loose structure, and alloy composition Pt/Au molar ratio is between 0.33 to 6, and mean diameter is at 20nm 40nm.Gained plation nano-particle analogue enztme solution has the highest oxidation catalytic activity to organic substrates 3,3 ', 5,5 ' tetramethyl benzidines in the case of hydrogen peroxide exists and be non-existent, shows similar peroxidase and oxidasic living features.And quasi-enzyme catalytic activity is strongly depend on Pt/Au ratio, the regulation and control to its mimetic enzyme catalysis activity can be realized according to this.
Description
Technical field
The present invention relates to a kind of plation nano-particle analogue enztme and its preparation method and application, belong to catalysis material and
Analytical chemistry field.
Background technology
Native enzyme relies on its high selectivity and the feature of high-efficiency catalytic activity, in biochemical field and food, agricultural, change
The aspects such as work play an important role.But the activity of native enzyme easily by temperature, chemical environment (such as pH value) etc. many because of
Element affects and degeneration, in addition complicated purification step and fancy price, all limits the application of native enzyme.This orders about numerous
Focus is placed on to find by scholar has a high-efficiency catalytic activity but the most stable catalyst, referred to as with native enzyme as
For artificial mimic enzyme.
In recent years, various types of nanostructureds are found to have the catalysis feature of similar native enzyme.Since the Chinese Academy of Sciences is raw
Yan Xiyun group of thing physics Institute reported first ferroferric oxide nano granules has the catalysis activity of class peroxidase
(L.Z.Gao,J.Zhuang,L.Nie,J.B.Zhang,Y.Zhang,N.Gu,T.Wang,J.Feng,D.L.Yang,
S.Perrett, X.Y.Yan, NatureNanotechnology, 2007,2,577-583), cerium oxide, cobalt oxide, copper sulfide,
The nano materials such as graphene oxide, gold, platinum are the most also found to have similar peroxidase, class oxidase, class hydrogen peroxide
The features such as enzyme.These nano-particle are simple, cheap due to preparation method, and are catalyzed the activity toleration to severe rugged environment
Relatively strong, become the nanostructured analogue enztme that a class is novel.Due to unique catalytic characteristics, nanostructured analogue enztme is in biological detection
Important using value is shown with fields such as immunoassays.
Noble metal nanometer material (such as Au Ag Pt Pd) shows the catalytic capability of uniqueness to different reaction systems,
The fields such as electro-catalysis, chemical and bio-sensing are all widely used.
Summary of the invention
One of the object of the invention is to provide a kind of dendroid plation nano-particle analogue enztme, and it has class oxidation simultaneously
Thing enzyme and the feature of class peroxidase activity, and activity is adjustable, can be used as oxide analogue enztme and Mimetic Peroxidase；
The two of the object of the invention are to provide method based on plation nano-particle class oxidase colorimetric detection sulfur hydrogen radical ion, the party
Method can be used for measuring sulfur hydrogen radical ion.
The technical scheme used is as follows:
Dendroid plation nano-particle analogue enztme, its plation nano-particle pattern is dendroid loose structure,
Alloy composition Pt/Au molar ratio is between 0.33 to 6, and mean diameter is at 20nm-40nm.
The preparation method of above-mentioned analogue enztme, comprises the following steps:
Gold chloride and potassium chloroplatinate are mixed in water, adds ascorbic acid (AA) vibration and shake up, 25-35 DEG C of isothermal reaction
4～5h, reaction adds cetyl trimethylammonium bromide (CTAB) after terminating, gained precipitate is centrifuged, wash, be scattered in from
In sub-water, obtain plation nano-particle mimetic enzyme solution.
By such scheme, gold is 3:1～1:10 with the ratio of the amount of the material of platinum element.
By such scheme, ascorbic acid mole be gold, 2～10 times of platinum element mole sum.
By such scheme, reaction temperature is 30 DEG C, and the response time is 4.5h.
By such scheme, ascorbic acid concentrations is 0.1M.
By such scheme, cetyl trimethylammonium bromide concentration is 0.1M.
By such scheme, time centrifugal, rotating speed is 12000rpm/min, duration 5-10min.
Above-mentioned analogue enztme is as oxide analogue enztme and the application of Mimetic Peroxidase.
Dendroid plation nano-particle analogue enztme as oxidase and Mimetic Peroxidase can respectively having and
Do not have during hydrogen peroxide, with TMB (TMB), chromogenic reaction occurs, be colourless become blue, have simultaneously
There are type oxide enzyme and the feature of class peroxidase activity, can be used as oxide analogue enztme and Mimetic Peroxidase, and
Its catalysis activity is strongly depend on the platinum/gold ratio of plation；
The colorimetric determination step of gold platinum nanoparticle mimic enzyme solution class oxidase active feature is as follows:
Take 3.0ml deionized water, be added thereto to successively 20 μ l 20mM TMB (TMB) and
25 μ l0.8mM plation nano-particle analogue enztme solution, then by above-mentioned solution mix homogeneously, gold platinum in described mixed solution
Alloy nanoparticle analogue enztme and 3,3 ', the amount of the material of 5,5 '-tetramethyl benzidine is than for 1:20.Place under room temperature (25 DEG C)
After 10-25 minute, solution i.e. be can be observed and become blue from colourless, these show that plation nano-particle analogue enztme is to water-soluble
Dissolved oxygen oxidation substrates TMB in liquid has the highest catalytic oxidation activity, shows the gold of the present invention
Platinum alloy nano-particle analogue enztme has similar oxidasic feature, can be as oxidase mimetic enzyme.
The colorimetric determination step of gold platinum nanoparticle mimic enzyme solution class peroxidase activity feature is as follows:
Take 3.0ml deionized water, add 20 μ l 20mM TMB (TMB), 20 μ the most wherein
l 0.1M H2O2With 25 μ l 0.8mM plation nano-particle analogue enztme solution, then by above-mentioned solution mix homogeneously, described
Plation nano-particle analogue enztme, hydrogen peroxide and 3,3 ' in mixed solution, the amount ratio of the material of 5,5 '-tetramethyl benzidine
For 1:100:20.After placing 10-25 minute under room temperature (25 DEG C), solution i.e. be can be observed and become blue from colourless, these show
Hydrogen peroxide oxidation substrate 3,3 ', 5,5 '-tetramethyl benzidine is had the highest catalysis to live by plation nano-particle analogue enztme
Property, show that the plation nano-particle analogue enztme of the present invention has the feature of similar peroxidase, can be as peroxide
Analogue enztme.
The application of above-mentioned analogue enztme colorimetric detection sulfur hydrogen radical ion concentration height.
The two of the object of the invention are that the inhibitory action utilizing sulfur hydrogen radical ion pair plation nano-particle analogue enztme realizes
Colorimetric detection to sulfur hydrogen radical ion, utilizes gold platinum nanoparticle mimic enzyme solution to the colorimetric determination step of sulfur hydrogen radical ion such as
Under:
Take 3.0ml deionized water, add 20 μ l 20mM TMB (TMB), 25 μ the most wherein
L 0.8mM plation nano-particle analogue enztme solution and concentration are (0.83-10 × 10-6MM) sodium hydrosulfide, then
By above-mentioned solution mix homogeneously；The detection to sulfate ion is realized by colorimetry or ultraviolet-visible absorption spectroscopy.Due to sulfur
The inhibitory action of hydrogen radical ion pair plation nano-particle mimetic enzyme catalysis activity, it can be observed that solution colour is along with sulfur hydrogen
Ion concentration raises, and solution colour is gradually become shallower as by dark blue, it is achieved the colorimetric detection to sulfur hydrogen radical ion.
The present invention, relative to prior art, has the beneficial effect that:
Being prepared for plation nano-particle analogue enztme solution by coreduction method, the method is simple to operate, efficient, heavy
Renaturation is high and reaction condition is gentle, and agents useful for same is nontoxic, and the product and the by-product that produce in reaction are the most environmentally friendly.
Gained plation nano-particle analogue enztme solution hydrogen peroxide exist and non-existent in the case of to organic end
Thing TMB has the highest oxidation catalytic activity, shows similar peroxidase and oxidasic
Living features.
The quasi-enzyme catalytic activity of gained plation is strongly depend on Pt/Au ratio, can realize according to this its mimetic enzyme catalysis
The regulation and control of activity.
Gained plation nano-particle analogue enztme has the highest under strong acid, highly basic, high salt concentration and hot conditions
Chemical stability, have potential as a kind of novel analogue enztme in fields such as immunoassay, biological detection and clinical diagnosises
Using value.
Accompanying drawing explanation
Fig. 1: different proportion plation nano-particle and the transmission electron microscope photo of Pt nanoparticle；
The uv absorption spectra of Fig. 2: different proportion plation nano-particle mimetic enzyme catalysis TMB oxidation.
Fig. 3: the kinetic curve of different proportion plation nano-particle class oxidase catalyzed TMB oxidation reaction.
The kinetic curve of Fig. 4: different proportion plation nano-particle class peroxidase catalysis TMB oxidation reaction.
Fig. 5: different proportion plation nano-particle class oxidase and class peroxidase activity correlation curve.
Fig. 6: different proportion plation nano-particle measures the concentration-colour developing rate profile of sulfur hydrogen radical ion.
Detailed description of the invention
For ease of understanding the present invention, the present invention is exemplified below.Those skilled in the art it will be clearly understood that described embodiment only
It is adapted to assist in and understands the present invention, be not construed as the concrete restriction to the present invention.
Embodiment 1
The preparation of Au nano-particle analogue enztme:
Take 2mL deionized water, be added thereto to tetra chlorauric acid aqueous solution that 16.7 μ L concentration are 24mM and ascorbic acid is molten
Liquid, vibration shakes up and is placed in 30 DEG C of thermostat water baths reaction 4.5 hours.It is 0.1M ten that reaction adds 0.1ml concentration after terminating
Six alkyl trimethyl bromination aqueous solutions (preparation is placed in 30 DEG C of constant temperature), deionized water wash constant volume obtains plation nanometer
Particle simulation enzymatic solution.Gold chloride in described solution and the amount ratio=0.4:4 of the material of ascorbic acid.Described plation is received
The concentration of rice grain analogue enztme solution is 0.8mM.
Embodiment 2
Au3Pt1The preparation (Pt/Au ratio is 0.33) of alloy nanoparticle analogue enztme:
Taking 2mL deionized water, being added thereto to 12.5 μ L concentration is that the tetra chlorauric acid aqueous solution of 24mM, 5.89 μ L concentration are
The potassium chloroplatinate aqueous solution of 17mM and ascorbic acid solution, vibration shakes up and is placed in 30 DEG C of thermostat water baths reaction 4.5 hours.
It is 0.1M cetyl trimethyl bromination aqueous solution (preparation is placed in 30 DEG C of constant temperature) that reaction adds 0.1ml concentration after terminating,
Deionized water wash constant volume obtains plation nano-particle analogue enztme solution.Gold chloride in described solution, potassium chloroplatinate, anti-
Amount ratio=the 0.3:0.1:4 of the material of bad hematic acid.The concentration of described plation nano-particle analogue enztme solution is 0.8mM.
Embodiment 3
Au1Pt1The preparation (Pt/Au ratio is 1) of alloy nanoparticle analogue enztme:
Taking 2mL deionized water, being added thereto to 8.3 μ L concentration is that the tetra chlorauric acid aqueous solution of 24mM, 17.7 μ L concentration are
The potassium chloroplatinate aqueous solution of 17mM and ascorbic acid solution, vibration shakes up and is placed in 30 DEG C of thermostat water baths reaction 4.5 hours.
It is 0.1M cetyl trimethyl bromination aqueous solution (preparation is placed in 30 DEG C of constant temperature) that reaction adds 0.1ml concentration after terminating,
Deionized water wash constant volume obtains plation nano-particle analogue enztme solution.Tetra chlorauric acid in described solution, potassium chloroplatinate,
Amount ratio=the 0.2:0.2:4 of the material of ascorbic acid.The concentration of described plation nano-particle analogue enztme solution is 0.8mM.
Embodiment 4
Au2Pt3The preparation (Pt/Au ratio is 1.5) of alloy nanoparticle analogue enztme:
Taking 2mL deionized water, being added thereto to 6.67 μ L concentration is that the tetra chlorauric acid aqueous solution of 24mM, 14.1 μ L concentration are
The potassium chloroplatinate aqueous solution of 17mM and ascorbic acid solution, vibration shakes up and is placed in 30 DEG C of thermostat water baths reaction 4.5 hours.
It is 0.1M cetyl trimethyl bromination aqueous solution (preparation is placed in 30 DEG C of constant temperature) that reaction adds 0.1ml concentration after terminating,
Deionized water wash constant volume obtains plation nano-particle analogue enztme solution.Tetra chlorauric acid in described solution, potassium chloroplatinate,
Amount ratio=the 0.16:0.24:4 of the material of ascorbic acid.The concentration of described plation nano-particle analogue enztme solution is
0.8mM。
Embodiment 5
Au1Pt3The preparation (Pt/Au ratio is 3) of alloy nanoparticle analogue enztme:
Taking 2mL deionized water, being added thereto to 4.17 μ L concentration is that the tetra chlorauric acid aqueous solution of 24mM, 17.6 μ L concentration are
The potassium chloroplatinate aqueous solution of 17mM and ascorbic acid solution, vibration shakes up and is placed in 30 DEG C of thermostat water baths reaction 4.5 hours.
It is 0.1M cetyl trimethyl bromination aqueous solution (preparation is placed in 30 DEG C of constant temperature) that reaction adds 0.1ml concentration after terminating,
Deionized water wash constant volume obtains plation nano-particle analogue enztme solution.Tetra chlorauric acid in described solution, potassium chloroplatinate,
Amount ratio=the 0.1:0.3:4 of the material of ascorbic acid.The concentration of described plation nano-particle analogue enztme solution is 0.8mM.
Embodiment 6
Au1Pt6The preparation (Pt/Au ratio is 6) of alloy nanoparticle analogue enztme:
Taking 2mL deionized water, being added thereto to 2.38 μ L concentration is the tetra chlorauric acid aqueous solution of 24mM, 20.17 μ L concentration
For potassium chloroplatinate aqueous solution and the ascorbic acid solution of 17mM, vibration shakes up that to be placed in 30 DEG C of thermostat water baths reaction 4.5 little
Time.It is that (preparation is placed on 30 DEG C of constant temperature to 0.1M cetyl trimethyl bromination aqueous solution that reaction adds 0.1ml concentration after terminating
In), deionized water wash constant volume obtains plation nano-particle analogue enztme solution.Tetra chlorauric acid in described solution, chlorine platinum
Acid potassium, the amount ratio=0.057:0.343:4 of material of ascorbic acid.The concentration of described plation nano-particle analogue enztme solution
For 0.8mM.
Embodiment 7
The preparation of Pt nano-particle analogue enztme:
Take 2mL deionized water, be added thereto to potassium chloroplatinate aqueous solution that 23.5 μ L concentration are 17mM and ascorbic acid is molten
Liquid, vibration shakes up and is placed in 30 DEG C of thermostat water baths reaction 4.5 hours.It is 0.1M ten that reaction adds 0.1ml concentration after terminating
Six alkyl trimethyl bromination aqueous solutions (preparation is placed in 30 DEG C of constant temperature), deionized water wash constant volume obtains plation nanometer
Particle simulation enzymatic solution.Potassium chloroplatinate in described solution and the amount ratio=0.4:4 of the material of ascorbic acid.Described pure platinum nanometer
The concentration of particle simulation enzymatic solution is 0.8mM.
Accompanying drawing 1 is the transmission electron microscope figure of the plation nanostructured of different Pt/Au ratios in embodiment.Pure Au
Irregular and nanometer spherical granule (Fig. 1 a) of surface smoother for pattern, along with Pt2+Addition, be 0.33 at Pt/Au ratio
Time, particle surface becomes uneven (Fig. 1 b), when Pt/Au ratio is 1, it can be seen that have dendritic particles to generate (Fig. 1 c), by
Cumulative big Pt/Au ratio, as Pt/Au=3 (Fig. 1 d), during Pt/Au=6 (Fig. 1 e), dendroid pattern is more and more obvious, and porous knot
It is more visible that structure also becomes ratio, and when especially Pt/Au is 6, this porous branch pattern becomes more tight, with pure Pt (Fig. 1 f) shape
Looks are close.AuPt nano particle diameter is more and more uniform.AuPt nano particle diameter is added up by we, and Pt/Au ratio is more
Greatly, grain diameter is the biggest, and change of size is from 20nm to 40nm.When Pt/Au ratio is 1, the chi of plation nano-particle
Very little particle diameter is 23.6 ± 2.3nm.
The colorimetric determination step of the class enzymatic activity of plation nano-particle analogue enztme solution is as follows:
Class oxidase characteristic test: take 3.0ml deionized water, is added thereto to 20 μ l 20mM 3,3 ', 5,5 '-four successively
Methyl biphenyl amine (TMB) and 25 μ l 0.8mM plation nano-particle analogue enztme solution (embodiment 1～6 prepares), then will
Above-mentioned solution mix homogeneously, plation nano-particle analogue enztme and TMB in described mixed solution
The amount of material than for 1:20；After placing 10-25 minute under room temperature (25 DEG C), solution i.e. be can be observed and becomes blue from colourless,
These show that plation nano-particle analogue enztme is to dissolved oxygen oxidation substrates 3,3 ', the 5,5 '-tetramethyl biphenyl in aqueous solution
Amine has the highest catalysis activity, shows that the plation nano-particle analogue enztme of the present invention has similar oxidasic feature, can
As oxidase mimetic enzyme.
Class peroxidase characteristics is tested: takes 3.0ml deionized water, is added thereto to 20 μ l 20mM 3,3 ', 5 successively,
5 '-tetramethyl benzidine (TMB), 20 μ l 0.1M H2O2(implement with 25 μ l 0.8mM plation nano-particle analogue enztme solution
Example 1～7 prepares), then by above-mentioned solution mix homogeneously, plation nano-particle analogue enztme, peroxide in described mixed solution
Changing hydrogen and 3,3 ', the amount of the material of 5,5 '-tetramethyl benzidine ratio is for 1:100:20；Place 10-25 minute under room temperature (25 DEG C)
After, solution i.e. being can be observed and become blue from colourless, these show that plation nano-particle analogue enztme is to hydrogen peroxide oxidation
Substrate TMB has the highest catalysis activity, shows the plation nano-particle analogue enztme of the present invention
There is the feature of similar peroxidase, can be as Mimetic Peroxidase.
Accompanying drawing 2 is the uv absorption spectra of the golden platinum nanoparticle mimic enzyme catalysis TMB oxidation of different-alloy ratio.From
It can be seen that different-alloy ratio gold Pt nanoparticle (Pt/Au=0.33,1,3,6) is in TMB aqueous solution during 15min in figure
Absorption spectrum change.Compared with catalyst blank, during Pt/Au=0.33 at 650nm the characteristic absorption peak of TMB oxidation product by
Gradually occur.And along with Pt/Au is than increasing up to 1,3 or 6 time, color is deepened the most therewith, the feature of TMB oxidation product at 650nm
Absworption peak is obviously enhanced, corresponding with absorption spectrum.
Fig. 3 and 4 is respectively the variant ratio alloy nanoparticle of equivalent and is catalyzed in the presence of with and without hydrogen peroxide
TMB oxidation absorbance change at 650nm.When Pt/Au ratio is gradually increased, the class oxidase of plation nano-particle
The most gradually strengthen with class Catalyzed Synthesis By Peroxidase activity.Either carry out in aqueous solution or the aqueous solution containing hydrogen peroxide,
The oxidation rate of TMB is all proportional with Pt/Au ratio.
Fig. 5 be when with or without hydrogen peroxide TMB oxidation reaction rate, between reaction rate and AuPt alloying component in
Linear relationship, reaction rate increases than increasing along with Pt/Au, is being simply added into 0.67mM H2O2Time TMB oxidation reaction speed
Rate does not has H exactly2O2Twice.These results indicate that we can regulate its quasi-enzyme catalytic by changing plation composition
Ability.
Based on plation nano-particle class oxidase colorimetric detection sulfur hydrogen radical ion, its step is:
1) Pt/Au embodiment 4 prepared mixes with TMB solution, with purple than the alloy nanoparticle analogue enztme being 1.5
Outer visible absorption spectra instrument scanning kinetics model is every the absorption curve of a 2min solution of record, feature at record 650nm
The absorbance of absworption peak, monitors and calculates the oxidation rate of TMB over time by absorbance.
2) after Pt/Au embodiment 3 prepared mixes with TMB solution than the alloy nanoparticle analogue enztme being 1.5, point
Do not add the sodium hydrosulfide of 0.83,1.66,3.33,10 μMs, every with ultraviolet-visible absorption spectroscopy instrument scanning kinetics model
At the absorption curve of a 2min solution of record, the record 650nm absorbance of characteristic absorption peak, by absorbance in time
The oxidation rate of TMB is monitored and is calculated in change.
Based on Pt nanoparticle class oxidase colorimetric detection sulfur hydrogen radical ion, its step is:
3) platinum nanoparticle mimic enzyme embodiment 7 prepared mixes with TMB solution, uses ultraviolet-visible absorption spectroscopy instrument
Scanning kinetics model is the absorbance of characteristic absorption peak at the absorption curve of a 2min solution of record, record 650nm, logical
Cross the oxidation rate that absorbance is monitored over time and calculated TMB.
4), after platinum nanoparticle mimic enzyme embodiment 7 prepared mixes with TMB solution, be separately added into 3.3,6.7,
16.6, the sodium hydrosulfide of 33 μMs, the most molten every 2min record with ultraviolet-visible absorption spectroscopy instrument scanning kinetics model
The absorption curve of liquid, at record 650nm, the absorbance of characteristic absorption peak, is monitored over time by absorbance and calculates
The oxidation rate of TMB.
It is fast to the TMB oxidation catalysis of same concentrations to Pt and AuPt nano-particle respectively that accompanying drawing 6 show different HS-concentration
The impact of rate.Find that Pt and AuPt alloy has identical feature: HS-concentration is the highest, and TMB oxidation rate is the lowest.It is worth note
Meaning, the addition of Au makes AuPt alloying, and they are different from pure Pt to the ability of the monitoring of HS-.The monitoring to HS-of the AuPt alloy
Than Pt slow 4 times, 0.83 μM of SH-can almost completely inhibit the catalysis that TMB is aoxidized by plation nano-particle, and pure Pt presses down
TMB processed oxidation needs 3.3 μMs.By contrast, Pt nano-particle (3.3 μMs to 33 μMs) is than plation alloy (0.83 μM to 10 μ
M) detection to HS-has the widely range of linearity.These results indicate that we can optimize by changing alloying component
AuPt alloy is to quasi-enzyme catalytic activity and the sensitivity of inhibitor.
From the foregoing, it will be observed that dendroid plation nanoparticles solution prepared by the present invention, have similar peroxidase and
Similar oxidasic catalysis, can be as the Mimetic Peroxidase of a kind of novelty and oxide analogue enztme, based on its fermentoid
Catalytic characteristics, the invention provides the colorimetric estimation method of sulfur hydrogen radical ion.Plation nano-particle mould prepared by the present invention
Intending enzyme can replace peroxidase and oxidase to be widely applied in field acquirements such as immunoassay, biological detection and clinical diagnosises
Prospect, additionally, the preparation of the plation nano-particle analogue enztme of the present invention, method is simple, quickly, green non-pollution and cost
Cheap.
Claims (9)
1. a dendroid plation nano-particle analogue enztme, it is characterised in that described plation nano-particle pattern is tree
Dendritic loose structure, alloy composition Pt/Au molar ratio is between 0.33 to 6, and mean diameter is at 20nm-40nm.
2. the preparation method of dendroid plation nano-particle analogue enztme described in claim 1, it is characterised in that include following
Step:
Gold chloride and potassium chloroplatinate are mixed in water, add ascorbic acid (AA) vibration shake up, 25-35 DEG C of isothermal reaction 4～
5h, reaction adds cetyl trimethylammonium bromide (CTAB) after terminating, and gained precipitate is centrifuged, washs, is scattered in deionization
In water, obtain plation nano-particle mimetic enzyme solution.
3. the preparation method of dendroid plation nano-particle analogue enztme as claimed in claim 2, it is characterised in that gold and platinum
The ratio of the amount of the material of element is 3:1～1:10.
4. the preparation method of dendroid plation nano-particle analogue enztme as claimed in claim 2, it is characterised in that Vitamin C
Acid mole be gold, 2～10 times of platinum element mole sum.
5. the preparation method of dendroid plation nano-particle analogue enztme as claimed in claim 2, it is characterised in that reaction temperature
Degree is 30 DEG C, and the response time is 4.5h.
6. the preparation method of dendroid plation nano-particle analogue enztme as claimed in claim 2, it is characterised in that Vitamin C
Acid concentration is 0.1M；Cetyl trimethylammonium bromide concentration is 0.1M.
7. the preparation method of dendroid plation nano-particle analogue enztme as claimed in claim 2, it is characterised in that time centrifugal
Rotating speed is 12000rpm/min, duration 5-10min.
8. dendroid plation nano-particle analogue enztme described in claim 1 is simulated as oxide analogue enztme and peroxide
The application of enzyme.
9. described in claim 1 dendroid plation nano-particle analogue enztme colorimetric detection sulfur hydrogen radical ion concentration height should
With.
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