CN1082186C - Preparation of organic state biological sensor from organic aquogel fixed enzyme - Google Patents
Preparation of organic state biological sensor from organic aquogel fixed enzyme Download PDFInfo
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- CN1082186C CN1082186C CN 95119390 CN95119390A CN1082186C CN 1082186 C CN1082186 C CN 1082186C CN 95119390 CN95119390 CN 95119390 CN 95119390 A CN95119390 A CN 95119390A CN 1082186 C CN1082186 C CN 1082186C
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Abstract
The present invention relates to a method for preparing a biosensor by using organic hydrogel immobilized enzymes. When the biosensor is prepared by the method, after enzymes, medium bodies, 0.1 to 5% surface active agent and 6 to 20% water solution of polyvinyl alcohol are evenly mixed, 60 to 85% dimethylformamide is added, and the mixed solution is dropwise coated on the surface of an electrode and gelatinized into films in 5 to 20 minutes at room temperature. The preparation process is rapid. The prepared biosensor has excellent properties: the biosensor has larger detection sensitivity to water-soluble substances and water-insoluble substances and is suitable for the biological detection in various media, such as waterless organic solvent, mixed oil-water solvent, water, etc.
Description
The invention belongs to organic hydrogel immobilized enzyme and prepare the method for biology sensor.
Organic phase biologic sensor, especially enzyme electrode have greatly been expanded Study on Biosensor and range of application, have a wide range of applications in fields such as organic chemical industry, bioengineering, medicine analysis, environmental protection.1988, people's reported first such as Hall organic phase enzyme electrode [G.F.Hall, D.I.Best, A.P.F.Tumer, EnzymeMicrob.Technol., 10 (1988) 543].Thenceforth so far, reported the organic phase enzyme electrode of many kinds, but all adopted conventional in the past water biology sensor process for fixation preparation, and must in organic solvent, add a certain amount of water, to guarantee the catalytic activity of enzyme.For example, J.Wang etc. are with Eastman AQ fixedly horseradish peroxidase and Tyrosinase, measure hydrogen peroxide and phenol in the nitrile, must add 4% water enzyme electrode and just have response [J.Wang, Y.Lin, L.Chen, Analyst, 118 (1993) 277] in nitrile.
Shaojun Dong, Yizhu Guo, Anal.Chem., 66 (1994) 3895; Shaojun Dong, Yizhu Guo, J.Chem.Soc., Chem.Commun., (1995) 483 report organic phase biologic sensor prepared from low temperature hydrogel enzymes, because carrier itself can provide little water environment for immobilised enzymes, need not to add water in medium, has realized the biological detection of hydrogen peroxide, phenolic compound in the real complete anhydrous organic solvent.But to water-insoluble compound, this method is powerless.
The objective of the invention is to add the organic solvent dimethyl formamide in the aqueous solution of surfactant and polyvinyl alcohol (PVA), gelation takes place at room temperature 5-20 minute in this mixed solution, prepares organic hydrogel.Preparation is during organic phase biologic sensor, with the aqueous solution of enzyme, mediator and surfactant and polyvinyl alcohol (PVA) even after, the dimethyl formamide of adding 60-75%, gel film forming under the room temperature.Preparation process is rapid, and it is short that the biology sensor of preparing reaches equilibration time, and response is fast.
The present invention is with percentage by weight: add the 60-75% dimethyl formamide in 0.5-5% surfactant and the 8-15% polyvinyl alcohol water solution, placed 5-20 minute under the room temperature, prepare organic hydrogel; During the preparation biology sensor, with enzyme, comprise peroxidase, cholesterol oxidase, Tyrosine oxidase, alcohol oxidase, alcohol dehydrogenase, laccase, CHE, choline oxidase, bilirubin oxidase, lipase etc., also can the aqueous solution of mediator iron hydrofining, ferrocene and derivant, organic dyestuff and surfactant and polyvinyl alcohol (PVA) is even, add dimethyl formamide, should mix drop and be coated onto the basal electrode surface, gel film forming under the room temperature; This biology sensor all has biocatalysis at chloroform, chlorobenzene, ethylene dichloride, dioxan, in the organic solvents such as nitrile, acetone, methyl alcohol, ethanol, normal butyl alcohol, n-octyl alcohol, cyclohexanone, tetrahydrofuran.
Owing to comprise ＂ alembroth ＂ dimethyl formamide and water in organic hydrogel of the present invention's preparation, it is to water-soluble substances, as hydrogen peroxide, phenolic compound etc., and water-insoluble materials, as organic superoxide, cholerythrin? cholesterol etc. all have bigger partition factor, thereby water-soluble and non-water soluble substance are all had bigger detection sensitivity.
The biology sensor of the present invention's preparation is compared with the various process for fixation of reporting in the past, and have following characteristic: (1) carrier can provide little water environment for immobilised enzymes; (2), comprise water and water-containing organic solvent at various media? stable existence in the complete anhydrous organic solvent; (3) all kinds of analytes all had bigger partition factor and detection sensitivity; (4) preparation process is rapid, need not specific installation.Therefore, this biology sensor wide adaptability both can also can be worked in absolute anhydrous organic phase in water or organic water phase.
Embodiment provided by the invention is as follows:
Embodiment 1: the horseradish peroxidase electrode.3 milligrams of horseradish peroxidases are dissolved in the aqueous solution of 50ml5% surfactant and 15% polyvinyl alcohol (PVA), add the dimethyl formamide of 100ml again, after mixing, this is mixed drop be coated in graphite electrode surface, following about 10 minutes gel film forming of room temperature.This enzyme electrode is to organic peroxides such as tert-butyl peroxide, peroxidating-2-butanone, the different acenes of hydrogen peroxide, and in the presence of the medialess body, all have sensitive response, and have excellent response performance: it is short to reach equilibration time, within 10 minutes; Response time is fast, within 1 minute; Measurement range is wide, 0-5mM.Can in organic solvents such as chloroform, chlorobenzene, alcohols, work.
Embodiment 2: the Tyrosinase electrode.1.5 milligrams of Tyrosinases are dissolved in the aqueous solution of 50ml 2% surfactant and 12% polyvinyl alcohol (PVA), add the 75ml dimethyl formamide again, after stirring, drip and to be coated onto the glass-carbon electrode surface, gel film forming about 15 minutes.This electrode performance: equilibration time, within 5 minutes; Response time, 0.5-1 minute.This enzyme electrode is measured chloroform, ethylene dichloride, the phenolic compound in the organic solvent such as nitrile.
Embodiment 3: the bilirubin oxidase electrode.2 milligrams of bilirubin oxidases are dissolved in the aqueous solution of 50ml 0.5% surfactant and 8% polyvinyl alcohol (PVA), add the 150ml dimethyl formamide again, after mixing, drip and to be coated onto the glass-carbon electrode surface, gel film forming about 8 minutes.This electrode is a mediator with the carboxylic acid ferrocene, measures the cholerythrin in chloroform, the dimethyl formamide.
Embodiment 4: the cholesterol oxidase electrode.4 milligrams of cholesterol oxidases are dissolved in the aqueous solution of 50ml 1% surfactant and 10% polyvinyl alcohol (PVA), add the 150ml dimethyl formamide again, after mixing, drip the glass-carbon electrode surface that is coated onto pre-modification platinum particulate, gel film forming about 8 minutes.This enzyme electrode is used for measuring the cholesterol of organic solvents such as chloroform, chlorobenzene.
Claims (3)
1. an organic hydrogel immobilized enzyme prepares the method for biology sensor, it is characterized in that with percentage by weight: in 0.5～5% surfactant and 8～15% polyvinyl alcohol water solutions, add enzyme or enzyme and mediator, after mixing, add the 60-75% dimethyl formamide, should mix drop and be coated onto the basal electrode surface, place the gel film forming under the room temperature 5～20 minutes.
2. organic hydrogel immobilized enzyme as claimed in claim 1 prepares method for biosensor, it is characterized in that described enzyme is peroxidase, cholesterol oxidase, Tyrosine oxidase, alcohol oxidase, alcohol dehydrogenase, laccase, CHE, choline oxidase, bilirubin oxidase or lipase.
3. organic according to claim 1 hydrogel immobilized enzyme prepares method for biosensor, it is characterized in that described mediator is the carboxylic acid ferrocene.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95119390 CN1082186C (en) | 1995-12-22 | 1995-12-22 | Preparation of organic state biological sensor from organic aquogel fixed enzyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95119390 CN1082186C (en) | 1995-12-22 | 1995-12-22 | Preparation of organic state biological sensor from organic aquogel fixed enzyme |
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| CN1152712A CN1152712A (en) | 1997-06-25 |
| CN1082186C true CN1082186C (en) | 2002-04-03 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 95119390 Expired - Fee Related CN1082186C (en) | 1995-12-22 | 1995-12-22 | Preparation of organic state biological sensor from organic aquogel fixed enzyme |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1067430C (en) * | 1997-09-15 | 2001-06-20 | 中国科学院长春应用化学研究所 | Method for preparing bio-sensor with embedding enzyme of collosol-gel |
| BRPI0619833A2 (en) * | 2005-12-16 | 2011-10-18 | Bayer Healthcare Llc | transdermal patch sensor assembly and methods of use |
| JP5572016B2 (en) * | 2009-08-04 | 2014-08-13 | シスメックス株式会社 | Tissue fluid extraction device, manufacturing method thereof, and tissue fluid analysis method using the device |
| CN105651841B (en) * | 2016-01-05 | 2018-03-06 | 宁夏医科大学 | The preparation method of the multiple stimulation response type hydrogel of multi-factor structure component film layer by layer |
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