Enzyme electrode and a method of determining concentration of an analyte in a sample solution

• US 5,118,404 A
• Filed: 02/27/1991
• Issued: 06/02/1992
• Est. Priority Date: 04/28/1989
• Status: Expired due to Term

• Current Assignee: NEC Corporation
• Original Assignee: NEC Corporation
• Inventors: Saito, Atsushi
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): Bell, Bruce F.
• Application Number:
  US07/660,911
• Time in Patent Office: 461 Days
• Field of Search: 204/153.12, 204/403, 357/25, 435/14, 435/288, 435/174, 435/817
• US Class Current: 205/778
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/003 Functionalisation
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

An enzyme electrode usable for determining high concentration of analyte in a sample solution is provided, by providing an enzyme-immobilized membrane in the electrode with pH buffer capacity. Durability of the electrode can be improved by using albumin crosslinked by glutaraldehyde as a permeation-restricted membrane in the electrode. Method of using the electrode can be simplified by introducing a stirring step, measuring outputs before and after the stirring step and utilizing difference of measured outputs.

Claims

1. An enzyme electrode comprising:

(a) an electrochemical transducer device for detecting variation of a phase boundary potential,

(b) an enzyme-immobilized membrane wherein an enzyme and carboxyl groups are immobilized, said carboxyl groups acting as a buffering agent against an acid produced by an enzyme reaction, and

(c) a permeation-restricted membrane having a restricted permeability to an analyte but having a good permeability to oxygen and being formed by material comprising an albumin crosslinked by glutaraldehyde;

said enzyme-immobilized membrane (b) and permeation-restricted membrane (c) being formed on the surface of said electrochemical transducer device (a) in the named order.

3. The enzyme electrode according to claim 1, wherein the enzyme-immobilized membrane (b) is formed by a layer (b1) of a material having pH buffer capacity and a layer (b2) of an immobilized enzyme, said layers (b1) and (b2) being formed on the surface of the electrochemical transducer device (a) in this order.

2. An enzyme electrode comprising:

(a) an electrochemical transducer device for detecting variation of a phase boundary potential,

(b) an enzyme-immobilized membrane formed by a layer (b1) of a material for immobilizing and containing carboxyl groups, said carboxyl groups acting as an agent for buffering against an acid produced by an enzyme reaction, and a layer (b2) of an immobilized enzyme, said layers (b1) and (b2) being formed on the surface of the electrochemical transducer device (a) in the named order, and

(c) a permeation-restriction membrane having a permeability restricted to an analyte but having a good permeability relative to oxygen and being formed by material comprising an albumin crosslinked by glutaraldehyde;

said enzyme-immobilized membrane (b) and permeation-restricted membrane (c) being formed on the surface of said electrochemical transducer device (a) in the named order.

4. The enzyme electrode according to claim 2, wherein the layer (b2) of an immobilized enzyme includes glucose oxidase and gluconolactose as immobilized enzyme.

5. A method of determining a concentration of an analyte in a sample solution by means of an enzyme electrode comprising:

(a) an electrochemical transducer device for detecting a variation of a phase boundary potential, (b) an enzyme-immobilized membrane wherein an enzyme and carboxyl groups are immobilized, said carboxyl groups acting as a buffering agent against an acid produced by an enzyme reaction, and (c) a permeation-restriction membrane having a restricted permeability to an analyte but having a good permeability relative to oxygen;

said enzyme-immobilized membrane (b) and permeation-restricted membrane (c) being formed on the surface of said electrochemical transducer device (a) in the named order, which method comprises the steps of;

(i) immersing said enzyme electrode in the sample solution for a predetermined time period,

(ii) stirring the sample solution, and

(iii) measuring a difference of electrode outputs between before the stirring and after the stirring, to determine a concentration of the analyte in the sample solution.