CN109248677A - Germanium dioxide nano enzyme and its Pesticides Testing application - Google Patents
Germanium dioxide nano enzyme and its Pesticides Testing application Download PDFInfo
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- CN109248677A CN109248677A CN201811199415.0A CN201811199415A CN109248677A CN 109248677 A CN109248677 A CN 109248677A CN 201811199415 A CN201811199415 A CN 201811199415A CN 109248677 A CN109248677 A CN 109248677A
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- 102000004190 Enzymes Human genes 0.000 title claims abstract description 76
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 76
- YBMRDBCBODYGJE-UHFFFAOYSA-N germanium dioxide Chemical compound O=[Ge]=O YBMRDBCBODYGJE-UHFFFAOYSA-N 0.000 title claims abstract description 72
- 239000000575 pesticide Substances 0.000 title claims abstract description 15
- 229940119177 germanium dioxide Drugs 0.000 title claims abstract description 11
- 238000012360 testing method Methods 0.000 title abstract description 5
- 229940088598 enzyme Drugs 0.000 claims abstract description 75
- 239000003987 organophosphate pesticide Substances 0.000 claims abstract description 20
- 102000003992 Peroxidases Human genes 0.000 claims abstract description 17
- 108040007629 peroxidase activity proteins Proteins 0.000 claims abstract description 17
- 230000000694 effects Effects 0.000 claims abstract description 14
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 11
- 108010022752 Acetylcholinesterase Proteins 0.000 claims abstract description 8
- 229940022698 acetylcholinesterase Drugs 0.000 claims abstract description 7
- 102000012440 Acetylcholinesterase Human genes 0.000 claims abstract 2
- 238000001514 detection method Methods 0.000 claims description 16
- 239000000243 solution Substances 0.000 claims description 13
- FHIVAFMUCKRCQO-UHFFFAOYSA-N diazinon Chemical compound CCOP(=S)(OCC)OC1=CC(C)=NC(C(C)C)=N1 FHIVAFMUCKRCQO-UHFFFAOYSA-N 0.000 claims description 10
- 230000031700 light absorption Effects 0.000 claims description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 8
- 239000008351 acetate buffer Substances 0.000 claims description 8
- -1 diamine salts Chemical class 0.000 claims description 6
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 5
- 230000015572 biosynthetic process Effects 0.000 claims description 5
- 229910052698 phosphorus Inorganic materials 0.000 claims description 5
- 239000011574 phosphorus Substances 0.000 claims description 5
- 238000003786 synthesis reaction Methods 0.000 claims description 5
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 229960004373 acetylcholine Drugs 0.000 claims description 3
- 239000012670 alkaline solution Substances 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical group CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 claims description 2
- KKUKTXOBAWVSHC-UHFFFAOYSA-N Dimethylphosphate Chemical compound COP(O)(=O)OC KKUKTXOBAWVSHC-UHFFFAOYSA-N 0.000 claims description 2
- 229910019142 PO4 Inorganic materials 0.000 claims description 2
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 claims description 2
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 claims description 2
- 238000007689 inspection Methods 0.000 claims description 2
- 229960004623 paraoxon Drugs 0.000 claims description 2
- WYMSBXTXOHUIGT-UHFFFAOYSA-N paraoxon Chemical compound CCOP(=O)(OCC)OC1=CC=C([N+]([O-])=O)C=C1 WYMSBXTXOHUIGT-UHFFFAOYSA-N 0.000 claims description 2
- LCCNCVORNKJIRZ-UHFFFAOYSA-N parathion Chemical compound CCOP(=S)(OCC)OC1=CC=C([N+]([O-])=O)C=C1 LCCNCVORNKJIRZ-UHFFFAOYSA-N 0.000 claims description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 2
- 239000010452 phosphate Substances 0.000 claims description 2
- IITIZHOBOIBGBW-UHFFFAOYSA-N 3-ethyl-2h-1,3-benzothiazole Chemical compound C1=CC=C2N(CC)CSC2=C1 IITIZHOBOIBGBW-UHFFFAOYSA-N 0.000 claims 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims 1
- 239000005864 Sulphur Substances 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract description 6
- 238000006243 chemical reaction Methods 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 230000002378 acidificating effect Effects 0.000 abstract description 2
- 238000012544 monitoring process Methods 0.000 abstract description 2
- 238000004445 quantitative analysis Methods 0.000 abstract description 2
- 230000008878 coupling Effects 0.000 abstract 1
- 238000010168 coupling process Methods 0.000 abstract 1
- 238000005859 coupling reaction Methods 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 239000000447 pesticide residue Substances 0.000 abstract 1
- 239000000463 material Substances 0.000 description 10
- 238000000034 method Methods 0.000 description 7
- 102100033639 Acetylcholinesterase Human genes 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- GFFIJCYHQYHUHB-UHFFFAOYSA-N 2-acetylsulfanylethyl(trimethyl)azanium Chemical compound CC(=O)SCC[N+](C)(C)C GFFIJCYHQYHUHB-UHFFFAOYSA-N 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000002086 nanomaterial Substances 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- QPLDLSVMHZLSFG-UHFFFAOYSA-N Copper oxide Chemical compound [Cu]=O QPLDLSVMHZLSFG-UHFFFAOYSA-N 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 229960001231 choline Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- GNTDGMZSJNCJKK-UHFFFAOYSA-N divanadium pentaoxide Chemical compound O=[V](=O)O[V](=O)=O GNTDGMZSJNCJKK-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- NUJOXMJBOLGQSY-UHFFFAOYSA-N manganese dioxide Chemical compound O=[Mn]=O NUJOXMJBOLGQSY-UHFFFAOYSA-N 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000011896 sensitive detection Methods 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- UXAMZEYKWGPDBI-UHFFFAOYSA-N C(CCCCCCCCCCCCCCC)Br(C)(C)C Chemical compound C(CCCCCCCCCCCCCCC)Br(C)(C)C UXAMZEYKWGPDBI-UHFFFAOYSA-N 0.000 description 1
- XMWRBQBLMFGWIX-UHFFFAOYSA-N C60 fullerene Chemical compound C12=C3C(C4=C56)=C7C8=C5C5=C9C%10=C6C6=C4C1=C1C4=C6C6=C%10C%10=C9C9=C%11C5=C8C5=C8C7=C3C3=C7C2=C1C1=C2C4=C6C4=C%10C6=C9C9=C%11C5=C5C8=C3C3=C7C1=C1C2=C4C6=C2C9=C5C3=C12 XMWRBQBLMFGWIX-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 206010008190 Cerebrovascular accident Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- 208000006011 Stroke Diseases 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 108010012715 Superoxide dismutase Proteins 0.000 description 1
- WGLPBDUCMAPZCE-UHFFFAOYSA-N Trioxochromium Chemical compound O=[Cr](=O)=O WGLPBDUCMAPZCE-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 239000010405 anode material Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- CETPSERCERDGAM-UHFFFAOYSA-N ceric oxide Chemical compound O=[Ce]=O CETPSERCERDGAM-UHFFFAOYSA-N 0.000 description 1
- 229910000422 cerium(IV) oxide Inorganic materials 0.000 description 1
- 229910000423 chromium oxide Inorganic materials 0.000 description 1
- UBEWDCMIDFGDOO-UHFFFAOYSA-N cobalt(II,III) oxide Inorganic materials [O-2].[O-2].[O-2].[O-2].[Co+2].[Co+3].[Co+3] UBEWDCMIDFGDOO-UHFFFAOYSA-N 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- OMZSGWSJDCOLKM-UHFFFAOYSA-N copper(II) sulfide Chemical compound [S-2].[Cu+2] OMZSGWSJDCOLKM-UHFFFAOYSA-N 0.000 description 1
- 239000011162 core material Substances 0.000 description 1
- 229910052955 covellite Inorganic materials 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- OCLWHYOULMTOJB-UHFFFAOYSA-L disodium hydrogen carbonate hydrate Chemical compound O.[Na+].[Na+].OC(O)=O.[O-]C([O-])=O OCLWHYOULMTOJB-UHFFFAOYSA-L 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 229910003472 fullerene Inorganic materials 0.000 description 1
- 239000000417 fungicide Substances 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 229910021389 graphene Inorganic materials 0.000 description 1
- 229910002804 graphite Inorganic materials 0.000 description 1
- 239000010439 graphite Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000013384 organic framework Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 239000000052 vinegar Substances 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
- B01J23/14—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00 of germanium, tin or lead
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/44—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase
- C12Q1/46—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase involving cholinesterase
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- General Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention relates to nano-catalytics, analytical chemistry field, specifically include germanium dioxide (GeO2) nano enzyme and its Pesticides Testing application.GeO2Nano enzyme has the activity of peroxidase at acidic, can catalyzing hydrogen peroxide and organicvisualization reagent generation chromogenic reaction.The present invention utilizes the GeO being prepared2Nano enzyme and acetylcholinesterase coupling realize the quantitative analysis of organophosphorus pesticide.The invention can be applied to the fields such as pesticide residue analysis, biological medicine, environmental monitoring.
Description
Technical field
The present invention relates to analytical chemistry fields, specifically include germanium dioxide nano enzyme and its Pesticides Testing application.
Background technique
Nano enzyme is a kind of novel analogue enztme, due to its powerful catalytic activity and in bio-sensing, food technology, ring
Protect etc., which has, potentially to be had also been applied as focus of attention.Nano enzyme overcomes many disadvantages of native enzyme, such as expensive,
Easy in inactivation and condition of storage require harshness etc., produce to fields such as bio-sensing, immunoassay, cancer diagnosis and treatments huge
It influences.
Up to the present, a variety of inorganic nano materials (including Co3O4, CeO2, V2O5, CuO, MnO2, CuS, FeS, your gold
Category, carbon (fullerene, graphene), metallo-organic framework and its composite nanostructure) all to show Mimicry enzyme (including imitative
Peroxidase, imitative oxidizing ferment, imitative catalase, imitative superoxide dismutase etc.).However, not all metal oxidation
Object all has Mimicry enzyme.Germanium dioxide has not been reported with Mimicry enzyme.
GeO2It is one of chromium oxide, since its is low in cost, synthesis technology is simple, and environmental stability is good, extensive
For replacing graphite as anode material.Meanwhile GeO2It is a kind of important glass formers, can effectively adjusts glass
The physical and chemical performance of refractive index and glass material is widely used in optical fiber core material and the design of covering material.GeO2Nano junction
Structure material is with important application prospects in fields such as luminescent devices with its excellent optical property.But we not yet have found at present
GeO2Nano enzyme has Mimicry enzyme.
Organophosphorus pesticide (OP) is most widely used agricultural fungicides, is improving agricultural productive force and is improving product quality
Aspect plays an important role.But organophosphorus pesticide will lead to the mankind and generate a variety of diseases, such as senile dementia, Parkinson's disease
With apoplexy etc..Its reason is mainly the work that organophosphorus pesticide can largely inhibit acetylcholinesterase in human body (AChE)
Property, cause the excessive of acetylcholine to be enriched with and then cause related disease.Simultaneously as the high residue characteristic of organophosphorus pesticide and
There are serious excessive use or non-standard operations in agricultural production, cause the high-content of pesticide on agricultural product to remain, serious prestige
Coerce the health and balance of nature of the mankind.Therefore, highly sensitive detection platform is constructed, realizes the easy, quick, high of pesticide
Sensitive Detection has important practical application value.So far, such as high performance liquid chromatography (HPLC), gas chromatography mass spectrometry (GC-MS), electrification
Credit analysis and colorimetric etc. have been applied to the sensing detection of organophosphorus pesticide.But these methods often have lower sensitivity
With the longer operating time, special professional is needed, and dependent on expensive large-scale instrument.These factors seriously limit
Large-scale application of the above method in Detecting Pesticide field.In conclusion the GeO with Mimicry enzyme2Nano enzyme
There is novelty in place field, and have not been reported at present based on GeO2The detection of organic phosphorus pesticide of nano enzyme.
Summary of the invention
The object of the present invention is to provide a kind of germanium dioxide nano enzyme and its applications in terms of Pesticides Testing.
To achieve the above object, the technical solution adopted by the present invention are as follows:
A kind of nano enzyme material and its application, the nano enzyme material GeO2Nano enzyme has imitative peroxidase activity.
Preferably, the imitative peroxidase material GeO2Nano enzyme synthesis process are as follows:
(1) under stiring by GeO2Solid powder is dissolved in sodium hydroxide solution, then by suitable cetyl trimethyl
Ammonium bromide (CTAB) is dissolved under stiring in appropriate distilled water, and 12 h are stirred at room temperature in two solution respectively；(2) it is stirring
It is lower that CTAB solution is poured into GeO2In alkaline solution, continue to stir 8 h at room temperature；(3) mixture is transferred to reaction under high pressure
In kettle, in 100 DEG C of 56 h of holding, then cool to room temperature.Up to germanium dioxide (GeO2) nano enzyme.
It is further preferred, the imitative peroxidase material GeO2Nano enzyme can be used as imitative Catalyzed Synthesis By Peroxidase agent,
For the application based on imitative peroxidase activity.
It is still further preferably the imitative peroxidase material GeO2Nano enzyme is in acid condition as with peroxide
The catalyst of compound enzymatic activity, for carrying out qualitative/quantitative detection to organophosphorus pesticide, steps are as follows:
(1) will delay comprising the phosphate of organic phosphorus (0 50 ng/L) of acetylcholinesterase (400 mU/mL) and various concentration
Fliud flushing (7.0,10 mmol/L of pH) is incubated for 30 minutes at 37 DEG C；(2) take the 10 above-mentioned mixed liquors of μ L in containing a certain amount of
GeO described in acetylthiocholine (0.1 mmol/L), claim 132Nano enzyme (100 μ g/mL), organicvisualization reagent
(0.5 mmol/L), H2O2In the acetate buffer (4.0,100 mmol/L of pH) of (0.5 mmol/L), continue at 37 DEG C
It is incubated for 30 minutes；(3) variation of observation solution colour detects organicvisualization reagent oxygen using spectrophotometer to realize qualitative detection
The corresponding light absorption value of compound realizes quantitative detection.
It is still more preferably the GeO2Nano enzyme can be used for detecting organophosphorus pesticide, the organophosphorus pesticide
It can be the organophosphorus pesticides such as paraoxon, parathion, 2-dichloroethylk dimethyl phosphate, basudin；The organicvisualization reagent is the 2,2 '-connection bis- (3- of nitrogen-
Ethyl benzo thiazole phenanthroline -6- sulfonic acid) diamine salts (ABTS), o-phenylenediamine (OPD) or 3,3 ', 5,5 '-tetramethyl benzidines
(TMB).
Effect of the invention is:
1. the present invention has synthesized the GeO with peroxidase activity2Nano enzyme, and optimal pH is 4.0 or so, in acidity
(pH 3.5 5.0) shows the activity of peroxidase under the conditions of pH.
2. the GeO that the present invention synthesizes2Nano enzyme shows the property different from imitative peroxidase nano material has been reported
Matter --- stability is low: activity drastically reduces at the pH below or above pH 3.5 5.0, is being below or above pH 3.5
5.0 pH stability inferior is substantially reduced.
3. the present invention utilizes GeO2The imitative peroxidase activity of nano enzyme and low stability, by GeO2Nano enzyme is applied to
The qualitative quantitative analysis of organophosphorus pesticide is realized and is detected using qualitative/quantitative of the spectrophotometry to organophosphorus pesticide.
4. the present invention provides the materials for playing Mimicry enzyme, in Detecting Pesticide, food safety, environmental monitoring etc.
Aspect has broad application prospects.
Detailed description of the invention
Fig. 1 is GeO provided in an embodiment of the present invention2The transmission electron microscope figure of nano enzyme；
Fig. 2 is GeO provided in an embodiment of the present invention2Nano enzyme Mimicry enzyme effect picture；
Fig. 3 is GeO provided in an embodiment of the present invention2The pH effect of optimization figure of nano enzyme Mimicry enzyme；
Fig. 4 is GeO provided in an embodiment of the present invention2The pH stability analysis figure of nano enzyme Mimicry enzyme；
Fig. 5 is H provided in an embodiment of the present invention2O2Quantitative measurement standard working curve；
Fig. 6 is detection of organic phosphorus pesticide schematic diagram provided in an embodiment of the present invention；
Fig. 7 is the qualitative detection photo of organophosphorus pesticide basudin provided in an embodiment of the present invention；
Fig. 8 is the quantitative measurement standard working curve of organophosphorus pesticide basudin provided in an embodiment of the present invention.
Specific embodiment
Illustrate the contents of the present invention deeper into ground to become apparent from, will further enumerate some embodiments, but this hair below
It is bright to be not limited to cited embodiment.Specific experiment condition or method are such as not specified in the following example, by this field
Normal condition or method carry out.
Embodiment 1
Imitative peroxidase material GeO2The preparation of nano enzyme:
By 1.06 g GeO2Solid powder is added in 10 mL sodium hydroxide solutions, by the cetyl trimethyl bromine of 1.46 g
Change ammonium (CTAB) to be dissolved in 12 mL distilled water, 12 h are stirred at room temperature in two solution respectively.Then CTAB solution is poured into
GeO2In alkaline solution, 8 h are stirred at room temperature.Finally mixture is transferred in autoclave, in 100 DEG C of 56 h of holding, so
After be cooled to room temperature.Up to the germanium dioxide (GeO with triangular structure2) nano enzyme.Fig. 1 is to pass through transmission electron microscope
The GeO observed2The TEM of nano enzyme is imaged.
Embodiment 2
GeO2The Mimicry enzyme of nano enzyme is verified:
Experimental system a: catalystic converter system is to include H2O2The GeO that (0.15 mmol/L), above-described embodiment obtain2Nano enzyme
(100 μ g/mL), organicvisualization reagent TMB(0.5 mmol/L) acetate buffer (4.0,100 mmol/L of pH).In room
After reacting 30 minutes under warm (25 DEG C), the light absorption value in its 450 800 nm is detected using microplate reader；
It separately does two check experiments: GeO is not added in the catalystic converter system of one of control experiment b2Nano enzyme, with it is above-mentioned
Light absorption value is detected after reacting 30 minutes under experimental system similarity condition；The catalystic converter system of another control experiment c is GeO2
Nano enzyme (100 μ g/mL) at acetate buffer (4.0,100 mmol/L of pH), with above-mentioned experimental system similarity condition
Lower standing detected light absorption value after 30 minutes；
As shown in Fig. 2, experimental system a shows apparent peak, illustrate GeO2Nano enzyme has at pH 4.0 significantly to be imitated
The activity of oxide enzyme；Check experiment b near 650 nm without apparent peak, if illustrating without GeO2Nano enzyme makees catalyst for nothing
Significant reaction；Check experiment c, without apparent peak, illustrates that the peak of experimental system a is not GeO near 650 nm2Nano enzyme itself
Response peak.
Embodiment 3
GeO2The pH of nano enzyme Mimicry enzyme optimizes:
Catalystic converter system is to include H2O2(0.3 mmol/L), GeO2Nano enzyme (100 μ g/mL), organicvisualization reagent TMB
Buffer (pH 1.0 2.0, the glycine-HCI buffer of the different pH of (0.5 mmol/L)；PH 3.0 6.0, acetic acid-vinegar
Sour sodium buffer；PH 6.5 8.0, phosphate buffer；PH 9.0 10.0, Tris hydrochloride buffer；PH 11.0 12.0,
Sodium bicarbonate-sodium hydrate buffer solution).After being reacted 30 minutes under room temperature (25 DEG C), detected at its 650 nm using microplate reader
Light absorption value.As shown in figure 3, GeO2Nano enzyme shows the activity of peroxidase at acidic (pH 3.5 5.0), and
And optimal pH is 4.0 or so.
Embodiment 4
GeO2The pH stability of nano enzyme Mimicry enzyme:
Catalystic converter system is to include GeO2Buffer (the pH 1.0 12.0,10 of nano enzyme (100 μ g/mL), difference pH
Mmol/L), after being incubated for 1h under room temperature (25 DEG C), organicvisualization reagent TMB(0.5 mmol/L is added), H2O2(0.3 mmol/L)
And acetate buffer (4.0,100 mmol/L of pH).Continuation utilizes microplate reader after reacting 30 min under room temperature (25 DEG C)
Detect the light absorption value in its 650 nm.As shown in figure 4, GeO2For nano enzyme at 3.0 8.0 pH, Mimicry enzyme is most stable, and
Optimal pH is 4.0 or so.
Embodiment 5
H2O2Quantitative detection:
Catalystic converter system is the H comprising various concentration2O2(0.01 0.3 mmol/L), GeO2Nano enzyme (100 μ g/mL) has
Machine color developing agent TMB(0.5 mmol/L) acetate buffer (4.0,100 mmol/L of pH).It is reacted under room temperature (25 DEG C)
After ten minutes, the light absorption value in its 300 800 nm is detected using microplate reader.After deducting blank control by 650 nm light absorption values
Draw out H2O2Standard working curve.As shown in figure 5, the range of linearity 0.01 0.15 mmol/L, y=1.97x+0.05 (R2=
0.9955)。
Embodiment 6
The qualitative detection of basudin:
Acetylthiocholine can generate mercaptoacetyl choline under acetylcholine ester enzymatic, can be with GeO2Nanometer enzyme reaction generates
Ge3+, to make GeO2The Mimicry enzyme of nano enzyme disappears.Organophosphorus pesticide can inhibit the activity of acetylcholinesterase, thus nothing
Method generates mercaptoacetyl choline, will not inhibit GeO2The Mimicry enzyme of nano enzyme, to realize the quantitative inspection of organophosphorus pesticide
It surveys.Fig. 6 is the detection schematic diagram of organophosphorus pesticide.Catalystic converter system is comprising acetylcholinesterase (400 mU/mL) and not
With the phosphate buffer (7,10 mmol/L of pH) of the basudin (0 50 ng/L) of concentration.It is incubated for 30 minutes at 37 DEG C
Afterwards, take the above-mentioned mixed liquor of 10 μ L that a certain amount of acetylthiocholine (0.1 mmol/L), GeO is added2Nano enzyme (100 μ g/
ML), organicvisualization reagent TMB(0.5 mmol/L), H2O2(0.5 mmol/L), acetate buffer (4.0,100 mmol/ of pH
L).30 min are incubated at 37 DEG C, as shown in fig. 7, observation color change.Solution colour is successively deepened, and illustrates GeO2Nano enzyme
It can detecte organophosphorus pesticide.
Embodiment 7
The quantitative detection of basudin:
Catalystic converter system is the basudin (0 50 ng/L) comprising acetylcholinesterase (400 mU/mL) and various concentration
After phosphate buffer (7.0,10 mmol/L of pH) is incubated for 30 minutes at 37 DEG C, the above-mentioned mixed liquor of 10 μ L is taken to be added one
Quantitative acetylthiocholine (0.1 mmol/L), GeO2Nano enzyme (100 μ g/mL), organicvisualization reagent TMB(0.5 mmol/
L), H2O2(0.5 mmol/L), acetate buffer (4.0,100 mmol/L of pH).After reaction 30 minutes, examined using microplate reader
The light absorption value surveyed at its 650 nm simultaneously draws out basudin standard working curve.As shown in figure 8, the range of linearity 5 30 ng/L, y=
0.008x+0.002 (R2=0.997)。
Claims (5)
1. a kind of germanium dioxide (GeO2) nano enzyme, it is characterised in that: the GeO2Nano enzyme has imitative peroxidase activity.
2. germanium dioxide (GeO according to claim 12) nano enzyme, it is characterised in that synthesis step is as follows:
(1) under stiring by GeO2Solid powder is dissolved in sodium hydroxide solution, then by suitable cetyl trimethyl
Ammonium bromide (CTAB) is dissolved under stiring in appropriate distilled water, and 12 h are stirred at room temperature in two solution respectively；(2) it is stirring
It is lower that CTAB solution is poured into GeO2In alkaline solution, continue to stir 8 h at room temperature；(3) that mixture is transferred to high pressure is anti-
It answers in kettle, in 100 DEG C of 56 h of holding, then cools to room temperature to get germanium dioxide (GeO2) nano enzyme.
3. according to claim 1, one kind described in 2 is based on GeO2The application of nano enzyme, it is characterised in that: institute in claim 1,2
State GeO2Nano enzyme can be used as imitative Catalyzed Synthesis By Peroxidase agent, for the application based on imitative peroxidase activity.
4. one kind is based on GeO2The application of the detection of organic phosphorus pesticide of nano enzyme, it is characterised in that organophosphorus pesticide qualitative/quantitative inspection
The step of surveying application is as follows:
(1) phosphate-buffered of the basudin (0 50 ng/L) of acetylcholinesterase (400 mU/mL) and various concentration will be included
Liquid (7.0,10 mmol/L of pH) is incubated for 30 minutes at 37 DEG C；(2) take the 10 above-mentioned mixed liquors of μ L in containing a certain amount of sulphur
For GeO described in acetylcholine (0.1 mmol/L), claim 132Nano enzyme (100 μ g/mL), organicvisualization reagent (0.5
Mmol/L), H2O2In the acetate buffer (4.0,100 mmol/L of pH) of (0.5 mmol/L), continuation is incubated at 37 DEG C
30 minutes；(3) variation of observation solution colour utilizes spectrophotometer detection organicvisualization reagent oxide to realize qualitative detection
Corresponding light absorption value realizes quantitative detection.
5. according to claim 4 a kind of based on GeO2The application of the detection of organic phosphorus pesticide of nano enzyme, it is characterised in that:
The organophosphorus pesticide can be the organophosphorus pesticides such as paraoxon, parathion, 2-dichloroethylk dimethyl phosphate, basudin, the organic chromogenic
Agent is 3,3 ', 5,5 '-tetramethyl benzidines (TMB), o-phenylenediamine (OPD) or 2,2 '-connection bis- (the 3- ethyl benzo thiazole phenanthrolines-of nitrogen-
6- sulfonic acid) diamine salts (ABTS).
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