Convenient determination of trace lead in whole blood and other fluids

• US 5,368,707 A
• Filed: 06/07/1993
• Issued: 11/29/1994
• Est. Priority Date: 01/15/1992
• Status: Expired due to Term

• Current Assignee: ESA Biosciences, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Andcare Incorporated
• Inventors: Liang, Zhi-Wei, Zhao, Junguo, Henkens, Robert W., Wojciechowski, Marek, O'Daly, John P., Morris, Susan E.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): Bell, Bruce F.
• Application Number:
  US08/073,806
• Time in Patent Office: 540 Days
• Field of Search: 204/153.12, 204/153.1, 204/153.17, 204/403, 204/412, 204/415, 204/418, 436/74, 436/77
• US Class Current: 205/777.5
• CPC Class Codes: C12Q 1/003 Functionalisation
• CPC Class Codes: C12Q 1/004 mediator-assisted
• 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: Y10S 435/817 Enzyme or microbe electrode

Abstract

The invention relates to methods of determining micromolar levels of lead ion in various fluids, including blood. Detection of lead or other heavy metal ion concentrations as low as 1 μg/dL is achieved. The methods are adaptable to the detection of low levels of lead in whole blood, employing a novel separation and release of lead ion from lead chelating agents. The disclosed methods employ isocitrate dehydrogenase-based electrodes which are particularly suited for detecting nanomolar levels of lead.

Claims

1. A method of determining lead levels in blood, comprising:

admixing a blood sample with an agent specifically binding lead to form a lead complex;

displacing the lead with a metal ion to form an irreversible metal ion complex; and

detecting the displaced lead and determining the lead levels in blood by electrochemical measurement.

2. The method of claim 1 wherein the irreversible metal ion complex is formed by oxidizing the metal ion.

3. The method of claim 2 wherein the metal ion is oxidized from a plus two to a plus three oxidation state.

4. The method of claims 1 or 2 wherein the electrochemical determination is with a bioelectrode which consists essentially of an immobilized enzyme in contact with a conducting surface and wherein inhibition of the enzyme by lead ions detectably affects current generated from redox reactions catalyzed by the enzyme when the bioelectrode is coupled with a reference electrode.

5. The method of claim 4 wherein the enzyme is isocitrate dehydrogenase.

6. The method of claim 1 wherein the agent specifically binding lead is dimercaptosuccinic acid, ethylenediaminetetraacetic acid, methylethylenediaminetetraacetic acid, or ethylene glycol-bis-(β

-aminoethylether) N,N,N'"'"',N'"'"'-tetraacetic acid.

7. The method of claim 1 wherein the agent specifically binding lead is ethylenediamine tetraacetic acid.

8. The method of claim 1 wherein the displacing metal ion is cobalt(II) or iron(II).

9. The method of claim 1 wherein the displacing metal ion is in a plus two oxidation state.

10. The method of claim 1 wherein the displacing metal is cobalt (II).

11. The method of claim 10 wherein cobalt(II) is oxidized to cobalt(III).