Method for the determination of glycated hemoglobin

• US 20020172992A1
• Filed: 05/14/2002
• Published: 11/21/2002
• Est. Priority Date: 05/16/2001
• Status: Active Grant

• Current Assignee: Abbott Diabetes Care Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: Abbott Diabetes Care Incorporated (Abbott Laboratories Incorporated)
• Inventors: Heller, Adam
• Granted Patent: US 7,005,273 B2
• Time in Patent Office: Days
• US Class Current: 435/25
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: F01B 1/12 Separate cylinder-crankcase elements coupled together to form a unit
• CPC Class Codes: F01L 9/20 by electric means
• CPC Class Codes: F02B 75/20 with cylinders all in one line
• CPC Class Codes: G01N 2333/805 Haemoglobins; Myoglobins
• CPC Class Codes: G01N 33/723 Glycosylated haemoglobin
• CPC Class Codes: H01M 8/16 Biochemical fuel cells, i.e. cells in which microorganisms function as catalysts
• CPC Class Codes: Y02E 60/50 Fuel cells

Abstract

A method of determining the percentage of glycated hemoglobin in a blood sample is disclosed wherein at least one of the assay steps is performed electrochemically. The method includes determining the total amount of hemoglobin in a sample by electrochemically measuring, in an oxygen electroreduction reaction at a cathode, the amount of oxygen in the sample. Because the amount of oxygen dissolved in the sample is known, the total hemoglobin is determined by subtracting the amount of free oxygen from the total oxygen measured, recognizing the fast equilibrium Hb+O₂⇄HbO₂. This can be followed by determining the amount of glycated hemoglobin in the sample. The cathode reaction is accomplished by contacting the sample with an enzyme, the enzyme being a copper-containing enzyme having four copper ions per active unit. The family of these enzymes includes, for example, laccases and bilirubin oxidases.

Claims

1. A method for determining the percentage of glycated hemoglobin in a fluid sample comprising:

electrochemically determining the total amount of hemoglobin in a blood sample;

determining the amount of glycated hemoglobin in the sample; and

calculating the ratio or percentage of glycated hemoglobin to total hemoglobin.

2. The method of claim 1, wherein electrochemically determining the total amount of hemoglobin comprises:

contacting the sample in an electrochemical cell with an enzyme capable of determining the oxygen content of the sample; and

calculating the amount of hemoglobin from the oxygen content.

3. The method of claim 2, wherein the enzyme is a copper-containing enzyme.

4. The method of claim 2, wherein the enzyme is selected from the group consisting of laccase, bilirubin oxidase, and ascorbate oxidase.

5. The method of claim 1, wherein determining the amount of glycated hemoglobin is performed spectrophotometrically.

6. The method of claim 1, wherein determining the amount of glycated hemoglobin is performed electrochemically.

7. The method of claim 1, wherein determining the amount of glycated hemoglobin is performed by direct enzymatic action.

8. The method of claim 1, wherein determining the amount of glycated hemoglobin is performed by direct enzymatic reaction.

9. A method for determining the percentage of glycated hemoglobin to total hemoglobin in blood comprising:

electrochemically determining the total amount of hemoglobin in a blood sample;

separating the glycated hemoglobin from the sample to form a second sample;

determining the amount of glycated hemoglobin in the second sample; and

calculating the percentage of glycated hemoglobin to total hemoglobin in the sample.

10. The method of claim 9, wherein determining the amount of glycated hemoglobin in the second sample is done electrochemically.

11. The method of claim 9, wherein determining the amount of glycated hemoglobin in the second sample is done spectrophotometrically.

12. A method for determining the percentage of glycated hemoglobin to total hemoglobin in blood comprising:

electrochemically determining the total amount of hemoglobin in a blood sample by exposing the sample to an enzyme associated with a cathode, determining the amount of oxygen from an enzymatic redox reaction, and calculating the amount of hemoglobin from the amount of oxygen;

determining the amount of glycated hemoglobin in the sample; and

calculating the percentage of glycated hemoglobin to total hemoglobin in the sample.