Method and sensor electrode system for the electrochemical determination of an analyte or an oxidoreductase as well as the use of suitable compounds therefor

• US 5,122,244 A
• Filed: 02/04/1991
• Issued: 06/16/1992
• Est. Priority Date: 02/03/1990
• Status: Expired due to Term

• Current Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Original Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Inventors: Hoenes, Joachim, Schaeffler, Jurgen
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): Bell, Bruce F.
• Application Number:
  US07/650,265
• Time in Patent Office: 498 Days
• Field of Search: 204/153.12, 204/403
• US Class Current: 205/777.5
• CPC Class Codes: C12Q 1/004 mediator-assisted

Abstract

The subject matter of the invention is a method for the electrochemical determination of an analyte in the presence of an oxidoreductase and a reducible substance which transfers electrons which arise during the course of the determination reaction from the oxidoreductase onto an electrode and thus leads to a signal which is a measure for the analyte to be determined whereby the reducible substance is enzymatically reduced and oxidized at the electrode, which is characterized in that the substance which forms at the electrode by oxidation is different from the reducible substance used initially, as well as a corresponding sensor electrode system and the use of compounds suitable therefor. Finally new nitrosoaniline derivatives and a process for their production are also subject matter of the invention.

Claims

1. A method for the electrochemical determination of an analyte in the presence of an oxidoreductase and a reducible substance which transfers electrons which arise during the course of the determination reaction from the oxidoreductase onto an electrode and thus leads to a signal which is a measure for the analyte to be determined whereby the reducible substance is enzymatically reduced and oxidized at the electrode in an overall irreversible reaction, wherein the substance which forms at the electrode by oxidation is an oxidized form of the enzymatically reduced substance other than the reducible substance used initially.

2. The method of claim 1, wherein said substance which forms at the electrode by oxidation is subsequently reduced by said oxido-reductase.

3. The method of claim 1, wherein said reducible substance accepts electrons arising during the course of the determination reaction from the oxidoreductase, thereby forming an electron-rich aromatic amine.

4. The method of claim 3, wherein said reducible substance is selected from the group of compounds of the formula I

5. The method of claim 1, wherein the oxidoreductase is selected from the group consisting of an oxidase, a non-NAD(P)-dependent dehydrogenase and diaphorase.

6. A method for the electrochemical determination of an oxidoreductase in the presence of a corresponding enzyme substrate and a reducible substance which is capable of transferring electrons from the oxidoreductase onto an electrode and thus leads to a signal which is a measure for the enzyme to be determined, whereby the reducible substance is enzymatically reduced and oxidized at the electrode in an overall irreversible reaction, wherein the substance which forms by oxidation at the electrode is an oxidized form of the enzymatically reduced substance other than the reducible substance used initially.

7. A method for the electrochemical determination of an analyte, comprising

contacting a sample containing said analyte with an oxidoreductase and a reducible substance which transfers electrons which are generated by the oxidation of said analyte onto an electrode;

oxidizing said analyte with said oxidoreductase to produce a reduced enzyme;

enzymatically reducing said reducible substance by said reduced enzyme to produce a reduced substance and regenerate said oxidoreductase,

oxidizing said reduced substance at an electrode, thereby transferring electrons generated by the oxidation of said analyte to said electrode, wherein said oxidation of said reduced substance forms a substance which is different from said reducible substance, wherein the overall reaction is irreversible.

8. The method of claim 7, wherein said reducible substance is selected from the group consisting of N,N-bis-(2-hydroxyethyl) nitrosoaniline and N-(2-hydroxyethyl)-N-(2-(2-hydroxyethyoxy-4-nitrosoaniline.

9. A method for the electrochemical determination of an oxidoreductase, comprising

contacting a sample containing said oxidoreductase in the presence of a corresponding enzyme substrate with a reducible substance which transfers electrons which are generated by the oxidation of said enzyme substrate onto an electrode;

enzymatically reducing said reducible substance by said oxidoreductase to produce a reduced substance,

oxidizing said reduced substance at an electrode, thereby transferring electrons generated by the reduction of said reducible substance to said electrode, wherein said oxidation of said reduced substance forms a substance which is different from said reducible substance, wherein the overall reaction is irreversible.