Enzymatic electrode and its preparation method

• US 5,272,087 A
• Filed: 10/16/1990
• Issued: 12/21/1993
• Est. Priority Date: 04/20/1988
• Status: Expired due to Term

• Current Assignee: Centre National De La Recherche Scientifique, Gwent Sensors Ltd. (Sun Chemical Corporation)
• Original Assignee: Centre National De La Recherche Scientifique
• Inventors: El Murr, Nabil, Slilam, Mohamed
• Primary Examiner(s): ALEXANDER, LYLE
• Application Number:
  US07/598,660
• Time in Patent Office: 1,162 Days
• Field of Search: 435/817, 435/14, 435/25, 435/288, 435/291, 435/176, 435/177, 435/180, 435/181, 435/311, 435/9, 204/403, 204/415, 204/418, 204/290 R, 204/153.12, 204/402, 204/
• US Class Current: 204/403.04
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

An enzymatic electrode, having a homogeneous composition throughout, consists essentially of a uniform intimate admixture of a conducting powder with at least one immobilized enzyme, or with an immobilized enzyme and a mediator agent or a co-enzyme, in a matrix. The electrode is one which has a surface available for direct contact with a substrate without a permeable or semi-permeable intermediate membrane or membranes. The enzymatic electrode is prepared, e.g., by intimately admixing (a) a homogeneous paste or matrix (resulting from an intimate admixture of a conducting powder with a binder), (b) at least one enzyme and (c) an enzyme crosslinking solution, and then incorporating therein a mediator agent and/or a co-enzyme.

Claims

1. An enzymatic electrode in the form of a matrix comprising a substantially homogenous admixture of a conducting powder and a member selected from the group consisting of (a) at least one immobilized enzyme, (b) an immobilized enzyme and a mediator agent and (c) an immobilized enzyme and a co-enzyme, wherein neither the mediator agent nor the co-enzyme is crosslinked to the matrix.

2. An enzymatic electrode of claim 1 having an intimately combined enzymatic part and detection part said electrode having a surface available for direct contact with a substrate without a permeable or semi-permeable intermediate membrane or membranes.

3. Electrode according to claim 2, which is constructed and adaptable is of the disposable, once-only use, type.

4. An electrode according to claim 2, wherein the conducting powder is carbon powder or graphite powder.

5. An electrode according to claim 2, wherein the conducting powder is capable of forming a matrix and physically, confining the enzyme.

6. An electrode according to claim 2, which contains several enzymes immobilized in the matrix.

7. An electrode according to claim 2, wherein the matrix is contained in an inert sheath, from which it protrudes at one end to form the surface, which is the useful surface of the electrode.

8. An electrode according to claim 2, for the determination of glucose, wherein the immobilized enzyme is glucose oxidase and the conducting matrix also contains a mediator selected from the group consisting of p-ferrocenylaniline, nickelocene and benzoquinone.

9. An electrode according to claim 2, for the determination of sucrose, wherein the at least one immobilized enzyme is invertase and glucose oxidase, or invertase, glucose oxidase and mutarotase.

10. An electrode according to claim 2, for the determination of ethanol, wherein the immobilized enzyme alcohol dehydrogenase, which is combined with NAD⁺.

11. An electrode according to claim 1 constructed and adapted to have a defined surface area for contacting the substrate, the surface area being one which is removable friction or cutting means to expose another surface having the identical surface area.

12. An electrode according to claim 1 wherein the mediator agent is a member selected from the group consisting of ferrocene, a ferrocene derivative, nickelocene, a nickelocene derivative and benzoquinone.

13. An electrode according to claim 2, wherein the enzyme is fixed within the matrix of conducting material by means of at least one crosslinking agent.

14. An electrode according to claim 13, wherein the crosslinking agent is glutaraldehyde.

15. An electrode according to claim 1 wherein the matrix comprises a conducting paste which is an intimate mixture of the powder with from 10 to 40% by weight of inert binder relative to the weight of the conducting paste.

16. An electrode according to claim 15, wherein the binder is a hydrophobic binder.

17. An electrode according to claim 16, wherein the binder consists of paraffin oil, alpha-bromo-naphthalene or a silicone oil.

18. A process for the preparation of an enzymatic electrode as defined in claim 2 which comprises intimately mixing

a homogeneous paste or matrix resulting from the intimate admixture of a conducting powder with a binder;

at least one enzyme; and

an enzyme crosslinking solution;

and then incorporating a mediator agent and/or a co-enzyme therein.

19. A process according to claim 18, wherein a conducting powder capable of forming a matrix and at least one enzyme are intimately mixed with a mediator agent and/or a co-enzyme and the resulting mixture is subjected to mechanical forces, in order to obtain a solid matrix physically confirming the immobilized enzyme.

20. A process according to claim 19, wherein the mechanical forces are compression forces.