Polarographic method for measuring both analyte and oxygen with the same detecting electrode of an electroenzymatic sensor

• US 5,030,333 A
• Filed: 10/14/1986
• Issued: 07/09/1991
• Est. Priority Date: 09/13/1984
• Status: Expired due to Fees

• Current Assignee: Children's Hospital & Medical Center
• Original Assignee: Children's Hospital & Medical Center
• Inventors: Clark, Leland Jr.
• Primary Examiner(s): Nguyen, Nam X.
• Application Number:
  US06/918,628
• Time in Patent Office: 1,729 Days
• Field of Search: 204/1 T, 204/1 Y, 204/1 P, 204/1 E, 204/403, 204/415, 204/418, 204/153.1, 204/153.12, 204/153.16, 204/153.17, 435/817, 128/637, 128/639, 128/642, 128/635
• US Class Current: 205/777.5
• CPC Class Codes: A61B 5/14865 invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors
• CPC Class Codes: C12Q 1/005 involving specific analytes or enzymes including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: G01N 27/3271 Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood amperometry per se G01N27/49; aspects concerning the enzyme reagent C12Q1/001

Abstract

Novel polarographic (voltametric) methods for measuring in vivo or in vitro analytes in an animal tissue and oxygen in the sensor enzymatic reaction layer with an electroenzymatic sensor are disclosed. The new and vastly improved methods are directed to generating from the electroenzymatic sensor at a suitable voltage an anodic current derived from an anodically active product generated by enzymatic activity on an analyte for measuring the analyte and a cathodic current derived from reducing the oxygen in the sensor enzymatic reaction layer for measuring the oxygen. Additionally, the method contemplates switching the voltage between anodic and cathodic potentials to generate anodic and cathodic currents or applying the voltage at a fixed rate. Advantageously, the methods of this invention may be utilized with implanted sensors in an animal including a human. The sensors may be implanted in blood or non-blood sites. Finally, the methods of this invention further contemplate analyzing the type of conditions around the implanted sensor site to improve the reliability of the sensor as well as the detection of the analyte by the sensor.

Claims

1. A polarographic method for detecting both an analyte and oxygen with a detecting electrode of an electro-enzymatic sensor having at least two or more electrodes, said method comprises:

contacting the analyte with the detecting electrode wherein the detecting electrode includes an enzymatic reaction layer containing an enzyme for reacting with the analyte in the presence of oxygen to generate an anodically active product;

exposing the same detecting electrode having a first polarity to a voltage for generating an anodic current derived from the anodically active product generated from the reaction between the enzyme and the analyte in the presence of oxygen;

reversing the polarity of the same detecting electrode;

exposing the same detecting electrode having a polarity opposite said first polarity to a voltage for generating a cathodic current derived from electrochemically reducing the oxygen in the enzymatic reaction layer; and

detecting both the anodic current and the cathodic current generated from the same detecting electrode as a function of the analyte and the oxygen, respectively, in the same enzymatic reaction layer.

2. A method of claim 1 further comprising implanting the electroenzymatic sensor in or on any tissue in an animal for in vivo measurement of the substrate and oxygen.

3. A method of claim 2 wherein the animal tissue is a non-blood site.

4. A method of claim 3 wherein the non-blood site is selected from the class consisting of a subcutaneous, intramuscular, epihepatic, intraperitoneal or brain site.

5. A method of claim 2 further comprising detecting an in vivo condition around the sensor site comprising

administering to the animal by inhalation a gas wherein the gas dissolves in the animal tissue,

detecting the current generated by the dissolved gas, and

determining the duration of the current which functions as an indication of the in vivo condition around the sensor.

6. A method of claim 5 wherein the dissolved gas is selected from the class consisting of a hydrogen gas or an oxygen gas.

7. A method of claim 6 wherein an anodic current derived from the dissolved hydrogen gas is detected and a cathodic current derived from the dissolved oxygen gas is detected.

8. A method of claim 7 wherein the in vivo condition is blood flow.

9. A method of claim 1 wherein the analyte is selected from the class consisting of glucose or lactate.

10. A method of claim 9 wherein the enzyme is glucose oxidase when the analyte is glucose and lactate oxidase when the analyte is lactate.

11. In a polarographic method for detecting glucose and oxygen with an electroenzymatic sensor having at least two or more electrodes, the improvement comprises:

contacting the glucose with a detecting electrode of the electroenzymatic sensor wherein the detecting electrode includes an enzymatic reaction layer containing glucose oxidase for reacting with the glucose in the presence of oxygen to generate gluconic acid and hydrogen peroxide;

exposing the same detecting electrode having a first polarity to a voltage for generating an anodic current derived from the hydrogen peroxide produced from the reaction between the glucose and glucose oxidase in the presence of oxygen;

reversing the polarity of the same detecting electrode;

exposing the same detecting electrode having a polarity opposite said first polarity to a voltage for generating a cathodic current derived from electrochemically reducing the oxygen in he enzymatic reaction layer; and

detecting the anodic current and the cathodic current generated from the same detecting electrode as a function of the glucose and the oxygen, respectively, in the same enzymatic reaction layer.