Amperometric flow injection analysis biosensor for glucose based on graphite paste modified with tetracyanoquinodimethane

• US 5,378,332 A
• Filed: 04/14/1993
• Issued: 01/03/1995
• Est. Priority Date: 04/14/1993
• Status: Expired due to Fees

• Current Assignee: The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: THE GOVERNMENT OF THE UNITED STATES OF AMERICA, AS REPRESENTED BY THE SECRETARY OF COMMERCE
• Inventors: Pandey, Prem C.
• Primary Examiner(s): Gorgos, Kathryn
• Assistant Examiner(s): Bell, Bruce F.
• Application Number:
  US08/046,187
• Time in Patent Office: 629 Days
• Field of Search: 204/403, 204/409, 204/153.12, 204/291, 435/817, 128/635
• US Class Current: 205/777.5
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

A biosensor system using flow injection analysis (FIA) for the analysis of glucose in human serum. The system utilizes a sensor in which the enzyme glucose oxidase (GOD) is incorporated into graphite paste modified with tetracyanoquinodimethane (TCNQ). TCNQ acts as an efficient mediator for oxidation of the reduced enzyme at 200 mV vs Ag/AgCl. The sensor provides a linear response to glucose over a wide concentration range. The flow injection analysis system has a detection limit of 200 nmol glucose using a 100 μl sample and as been demonstrated to be stable after 800 consecutive sample injections.

Claims

1. An electrode for a biosensor which comprises a homogeneous composition of between about 8 to 15 weight percent of a redox enzyme, between about 28 to 35 weight percent of an electron-transfer mediator, between about 28 to 35 weight percent graphite powder, and between about 23 to 30 weight percent of a mineral oil.

2. An electrode for a biosensor according to claim 1, wherein said redox enzyme comprises glucose oxidase.

3. An electrode for a biosensor according to claim 2 wherein said electron-transfer mediator comprises tetracyanoquinodimethane.

4. An electrode for a biosensor according to claim 2 wherein said mineral oil is Nujol oil.

5. An electrode for a biosensor according to claim 2, wherein said mineral oil has a density of between about 0.83 and 0.85 g/mL.

6. An electrode for a biosensor according to claim 2, wherein said graphite powder has a particle size of less than 10 microns.

7. An electrode for a biosensor according to claim 1, wherein said composition is held in a support.

8. In a flow-injection analysis system comprising a pump, an electrochemical cell, a flow line connected between said pump and said electrochemical cell and a sample injector connected to said flow line, the improvement comprising an enzyme electrode for said electrochemical cell which comprises a homogeneous composition of between about 9 to 12 weight percent of a redox enzyme, between about 30 to 33 weight percent of an electron-transfer mediator, between about 30 to 33 weight percent graphite powder, and between about 24 to 28 weight percent of a mineral oil.

9. A flow-injection analysis system according to claim 8, wherein said redox enzyme comprises glucose oxidase.

10. A flow-injection analysis system according to claim 9, wherein said electron-transfer mediator comprises tetracyanoquinodimethane.

11. A flow-injection analysis system according to claim 9, wherein said mineral oil comprises Nujol oil.

12. A flow-injection analysis system according to claim 9, wherein said mineral oil has a density of between about 0.83 and 0.85 g/mL.

13. A flow-injection analysis system according to claim 12, wherein said mineral oil has a density of about 0.838 g/mL.

14. A flow-injection analysis system according to claim 9, wherein said graphite powder has a particle size of less than 10 microns.

15. A flow-injection analysis system according to claim 14, wherein said graphite powder has a particle size of between 1-2 microns.

16. A flow-injection analysis system according to claim 8, wherein said composition is held in a support.

17. A method of measuring glucose concentration in a sample which comprises:

providing a sample fluid containing glucose;

providing an electrochemical cell having a working electrode which comprises a homogeneous composition of between about 8 to 15 weight percent of glucose oxidase, between about 28 to 35 weight percent of tetracyanoquinodimethane, between about 28 to 35 weight percent graphite powder and between about 23 to 30 weight percent of mineral oil;

supplying said sample fluid to said electrochemical cell; and

measuring glucose concentration in said sample fluid by means of said electrochemical cell.

18. A method of measuring glucose concentration in a sample according to claim 17, wherein said electrochemical cell comprises a flow cell and said sample fluid is supplied to said electrochemical cell by means of a flow-injection system.

19. A method of measuring glucose concentration in a sample according to claim 17, wherein said sample comprises blood serum.