Enzyme electrode and method for determination of alcohol content using the same
First Claim
1. An enzyme electrode for amperometric measurement, comprising a conductive base material and an immobilized enzyme membrane or an immobilized enzyme layer, said immobilized enzyme membrane or immobilized enzyme layer comprised of a crosslinked reaction product of an alcohol oxidase solution containing alcohol oxidase, a crosslinking agent and reduced glutathione.
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Abstract
An enzyme electrode possessing an immobilized enzyme membrane or an immobilized enzyme layer composed by applying a solution containing alcohol oxidase and a crosslinking agent, in which the solution further contains reduced glutathione.
According to the invention it is possible to immobilize alcohol oxidase stably, and an excellent immobilized enzyme electrode for alcohol measurement is obtained. Furthermore, by this invention, a highly sensitive and stable measuring method is conducted quite easily.
17 Citations
18 Claims
- 1. An enzyme electrode for amperometric measurement, comprising a conductive base material and an immobilized enzyme membrane or an immobilized enzyme layer, said immobilized enzyme membrane or immobilized enzyme layer comprised of a crosslinked reaction product of an alcohol oxidase solution containing alcohol oxidase, a crosslinking agent and reduced glutathione.
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12. A method for determination of alcohol content in a sample comprising the steps of immersing an enzyme electrode in a sample solution, said enzyme electrode comprising a conductive base material and an immobilized enzyme membrane or an immobilized enzyme layer, said immobilized enzyme membrane or immobilized enzyme layer comprised of a crosslinked reaction product of an alcohol oxidase solution containing alcohol oxidase, a crosslinking agent and reduced glutathione, and measuring with said enzyme electrode the amount of oxygen consumed by the oxidation of alcohol catalyzed by said alcohol oxidase.
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13. A method for determination of alcohol content in a sample comprising the steps of immersing an enzyme electrode in a sample solution, said enzyme electrode comprising a conductive base material and an immobilized enzyme membrane or an immobilized enzyme layer, said immobilized enzyme membrane or immobilized enzyme layer comprised of a crosslinked reaction product of an alcohol oxidase solution containing alcohol oxidase, a crosslinking agent and reduced glutathione, and measuring with said enzyme electrode the amount of hydrogen peroxide produce by the oxidation of alcohol catalyzed by said alcohol oxidase.
- 14. A method for determination of alcohol content in a sample by amperometric measurement comprising the steps of immersing an enzyme electrode in a sample solution comprising a buffer solution and sodium azide, said enzyme electrode comprising a conductive base material and an immobilized enzyme membrane or an immobilized enzyme layer, said immobilized enzyme membrane or immobilized enzyme layer comprised of a crosslinked reaction product of an alcohol oxidase solution containing alcohol oxidase, reduced glutathione and a crosslinking agent, and measuring with said enzyme electrode either the amount of oxygen consumed or the amount of hydrogen peroxide produced by the oxidation of alcohol catalyzed by said alcohol oxidase.
- 16. A method for determination of alcohol content in a sample by amperometric measurement comprising the steps of immersing an enzyme electrode in a sample solution comprising a buffer solution and polyacrylic acid, said enzyme electrode comprising a conductive base material and an immobilized enzyme membrane or an immobilized enzyme layer, said immobilized enzyme membrane or immobilized enzyme layer comprised of a crosslinked reaction product of an alcohol oxidase solution containing alcohol oxidase, reduced glutathione and a crosslinking agent, and measuring with said enzyme electrode either the amount of oxygen consumed or the amount of hydrogen peroxide produced by the oxidation of alcohol catalyzed by said alcohol oxidase.
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18. A method for determination of alcohol content in a sample comprising the steps of immersing an enzyme electrode in a sample solution comprising a buffer solution and sodium azide, said enzyme electrode comprising a conductive base material and an immobilized enzyme membrane or an immobilized enzyme layer, said immobilized enzyme membrane or immobilized enzyme layer comprised of a crosslinked reaction product of an alcohol oxidase solution containing alcohol oxidase, a crosslinking agent and reduced glutathione, and measuring with said enzyme electrode the amount of hydrogen peroxide produced by the oxidation of alcohol catalyzed by the alcohol oxidase, wherein the concentration of sodium azide in the buffer solution is 0.1 μ
- M or more and 1 mM or less.
Specification