Perfluorosulfonic acid polymer-coated indicator electrodes

• US 4,604,182 A
• Filed: 01/29/1985
• Issued: 08/05/1986
• Est. Priority Date: 08/15/1983
• Status: Expired due to Term

• Current Assignee: Dade Behring, Inc. (Siemens AG)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Seago, James L.
• Primary Examiner(s): Niebling, John F.
• Assistant Examiner(s): Chapman, Terryence
• Application Number:
  US06/696,196
• Time in Patent Office: 553 Days
• Field of Search: 204/1 E, 204/1 T, 204/403, 204/415, 204/418, 435/288, 435/291
• US Class Current: 204/403.11
• CPC Class Codes: C12M 21/18 Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
• CPC Class Codes: C12M 41/32 of substances in solution
• CPC Class Codes: C12Q 1/005 involving specific analytes or enzymes including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence

Abstract

A method is described for the amperometric determination of the concentration in a biological sample of a constituent which is a substrate for an oxidase enzyme by (1) catalyzing the substrate with the enzyme, thereby consuming O₂ and producing H₂ O₂ and (2) measuring the O₂ consumption or H₂ O₂ production by means of an inert metal indicator electrode intimately coated with a thin film of a perfluorosulfonic acid polymer.

A reaction chamber is described which provides means for contacting the sample with the enzyme and means for making amperometric measurements utilizing inert metal indicator electrodes coated with a thin film of a perfluorosulfonic acid polymer.

Claims

1. A flow-through enzyme reactor for the amperometric determination of a constituent of interest contained in a liquid sample, comprising:

a first and a second end member, each having an internal chamber communicable with a source and a sink, respectively, of said liquid sample, said first and second chambers each having disposed therein a noble metal wire having coated thereon a thin film of a perfluorosulfonic acid polymer, said wires being externally connectable to electronic circuitry, said end members being made from an insulating material; and

a central member having an internal chamber adapted to contain an immobilized oxidase enzyme, said chamber in communication with said internal chambers of said end members, said central chamber being separated from each of said end chambers by a foraminous septum, said septum being capable of containing the immobilized enzyme within the central chamber, said central chamber being made from an inert, electrically conductive material.

2. The reactor of claim 1 wherein the noble metal wire is selected from the group consisting of platinum and gold.

3. The reactor of claim 2 wherein the noble metal wire is platinum.

4. The reactor of claim 1 wherein the thin film is less than about 0.005 inch.

5. The reactor of claim 1 wherein the thin film is about 0.001 inch.

6. The reactor of claim 1 wherein the perfluorosulfonic acid polymer has an equivalent weight of at least about 900.

7. The reactor of claim 6 wherein the perfluorosulfonic acid polymer has an equivalent weight of about 1100.

8. The reactor of claim 1 wherein the central member is made from an inert conductive material selected from the group consisting of graphite, wax-filled graphite, polymer-filled graphite, pyrolitic graphite and glassy carbon.

9. The reactor of claim 8 wherein the inert conductive material is glassy carbon.

10. The reactor of claim 1 wherein the noble metal wire is about 38 gauge.

11. The reactor of claim 1 wherein the foraminous septum is a nylon mesh.