CN1152712A - 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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- CN1152712A CN1152712A CN 95119390 CN95119390A CN1152712A CN 1152712 A CN1152712 A CN 1152712A CN 95119390 CN95119390 CN 95119390 CN 95119390 A CN95119390 A CN 95119390A CN 1152712 A CN1152712 A CN 1152712A
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Abstract
The present invention relates to a method for preparing biosensor. It is characterized by adopting the following steps: using enzyme, medium, 0.1-5% surfactant and 6-2% polyvinyl alcohol liquor; uniformly mixing them, adding 60-85% dimethylformamide; drop-a;;ying the mixed solution to the surface of electrode and forming gel film within 5-20 min. at room temp.. Its preparation process is quick, and the prepared biosensor possesses the excellent properties, and has the higher detection sensitivity for water soluble and non-water soluble material, and is applicable to biodetection of various media of absolute organic solvent, oil-water mixed solvent and 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.Turner, Enzyme Microb.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, Eastman AQ fixedly horseradish peroxidase and tyrosinase such as J.Wang are measured hydrogen peroxide and phenol in the acetonitrile, must in acetonitrile, add 4% water enzyme electrode and just have response [J.Wang, Y.Lin, L.Chen, Analyst, 118 (1993) 277].
Shaojun Dong, Yizhu Guo, Anal.Chem., 66 (1994) 3895; Shaojun Dong, YizhuGuo, 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 60-85%, 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 adds the 60-85% dimethyl formamide in 0.1-5% surfactant and 6-20% polyvinyl alcohol water solution, placed 5-20 minute under the room temperature, prepares 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 be with mediator, even as the potassium ferricyanide, ferrocene and derivant, organic dyestuff etc. with the aqueous solution of surfactant and polyvinyl alcohol (PVA), 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 in organic solvents such as chloroform, chlorobenzene, ethylene dichloride, dioxan, acetonitrile, 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, all have bigger partition factor as organic superoxide, cholerythrin, cholesterol etc., 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 stable existence in water and water-containing organic solvent, the complete anhydrous organic solvent at various media; (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 50ul 5% surfactant and 15% polyvinyl alcohol (PVA), the dimethyl formamide that adds 100ul 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 50ul 2% surfactant and 12% polyvinyl alcohol (PVA), add the 75ul 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 the phenolic compound in the organic solvents such as chloroform, ethylene dichloride, acetonitrile.
Embodiment 3: the bilirubin oxidase electrode.2 milligrams of bilirubin oxidases are dissolved in the aqueous solution of 50ul 0.5% surfactant and 8% polyvinyl alcohol (PVA), add the 150ul 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 50ul 1% surfactant and 10% polyvinyl alcohol (PVA), add the 150ul 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 (1)
1. an organic hydrogel immobilized enzyme prepares the method for biology sensor, it is characterized in that placing 5-20 minute under the room temperature to add the 60-85% dimethyl formamide in 0.1-5% surfactant and the 6-20% polyvinyl alcohol water solution, prepares 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 be with mediator, even as the potassium ferricyanide, ferrocene and derivant, organic dyestuff etc. with the aqueous solution of surfactant and polyvinyl alcohol (PVA), add dimethyl formamide, should mix drop and be coated onto the basal electrode surface, gel film forming under the room temperature; In the organic solvents such as this biology sensor is imitative at oxygen, chlorobenzene, ethylene dichloride, dioxan, acetonitrile, acetone, methyl alcohol, ethanol, normal butyl alcohol, n-octyl alcohol, cyclohexanone, tetrahydrofuran, all has biocatalysis.
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 |
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| Application Number | Priority Date | Filing Date | Title |
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| 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 true CN1152712A (en) | 1997-06-25 |
| CN1082186C 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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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN101947104A (en) * | 2005-12-16 | 2011-01-19 | 拜尔保健有限公司 | Analyte sensor assembly of transdermal and using method thereof |
| CN103800040A (en) * | 2009-08-04 | 2014-05-21 | 希森美康株式会社 | Device for interstitial fluid extraction, production process thereof and analyzing process of interstitial fluid using the device |
| CN105651841A (en) * | 2016-01-05 | 2016-06-08 | 宁夏医科大学 | Method for preparing multiple-structure multi-stimuli-responsive hydrogel layer-by-layer assembled film |
-
1995
- 1995-12-22 CN CN 95119390 patent/CN1082186C/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN101947104A (en) * | 2005-12-16 | 2011-01-19 | 拜尔保健有限公司 | Analyte sensor assembly of transdermal and using method thereof |
| CN101947104B (en) * | 2005-12-16 | 2013-02-06 | 拜尔保健有限公司 | Transdermal analyte sensor assembly and methods of using the same |
| US8504131B2 (en) | 2005-12-16 | 2013-08-06 | Bayer Healthcare Llc | Transdermal analyte sensor assembly and methods of using the same |
| CN103800040A (en) * | 2009-08-04 | 2014-05-21 | 希森美康株式会社 | Device for interstitial fluid extraction, production process thereof and analyzing process of interstitial fluid using the device |
| CN103800040B (en) * | 2009-08-04 | 2016-06-01 | 希森美康株式会社 | Tissue juice extraction device, its manufacture method |
| CN105651841A (en) * | 2016-01-05 | 2016-06-08 | 宁夏医科大学 | Method for preparing multiple-structure multi-stimuli-responsive hydrogel layer-by-layer assembled film |
| 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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| CN1082186C (en) | 2002-04-03 |
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