Whole blood hemolysis sensor

• US 9,658,181 B2
• Filed: 03/10/2014
• Issued: 05/23/2017
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: Instrumentation Lab Co (Instrumentation Lab)
• Original Assignee: Instrumentation Lab Co (Instrumentation Lab)
• Inventors: D'Orazio, Paul, Balasubramanian, Shankar
• Primary Examiner(s): Dinh, Bach
• Application Number:
  US14/202,398
• Publication Number:
  US 20140262831A1
• Time in Patent Office: 1,170 Days
• Field of Search: 20440301-40315, 2057775, 205779, 205778, 205792, 205793
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• 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 1/26 involving oxidoreductase
• CPC Class Codes: G01N 27/327 Biochemical electrodes , e.g. electrical or mechanical details for in vitro measurements
• 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
• CPC Class Codes: G01N 27/3272 Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels optical biosensors G01N33/52
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48

Abstract

The present invention pertains to a hemolysis sensor, a hemolysis sensor system and methods of utilizing the hemolysis sensor or hemolysis sensor system to monitor or detect hemolysis in a sample, such as a whole blood sample, a plasma sample, a serum sample or hemolyzed blood. The hemolysis sensor responds to extracellular hemoglobin levels, for example, extracellular hemoglobin in a whole blood sample as a method for detecting hemolysis in whole blood.

Claims

1. A method for detecting hemolysis in a whole blood sample, comprising:

(i) introducing said whole blood sample to an electrochemical sensor, said electrochemical sensor comprising an oxidoreductase enzyme capable of generating hydrogen peroxide and a hydrophilic outer membrane comprising a thickness in a range of about 0.1 μ

m to about 50 μ

m, said membrane thickness adapted to enhance efflux of said hydrogen peroxide, followed by;

(ii) detecting an electrochemical signal generated by said hydrogen peroxide in the presence of hemoglobin (Hb(Fe²⁺)), wherein a decrease of the electrochemical signal in a range of about 4% to about 50% as compared to a standard non-hemolyzed whole blood sample, is indicative of said hemolysis in the whole blood sample.

2. The method of claim 1 wherein the oxidoreductase enzyme comprises a glucose oxidase.

3. The method of claim 1 wherein the oxidoreductase enzyme comprises a lactate oxidase.

4. The method of claim 1 wherein, the oxidoreductase enzyme is a creatininase.

5. The method of claim 1 wherein the current generated at the working electrode is lower in the presence of said hemoglobin (Hb(Fe²⁺)) as compared to the current generated in the absence of said hemoglobin (Hb(Fe²⁺)).

7. The method of claim 1 wherein the electrochemical signal is a detectable electrical current.

8. The method of claim 1 wherein said outer hydrophilic membrane comprises a hydrogel.

9. The method of claim 8 wherein the hydrogel comprises water content ranging from about 0.1% to about 100%.

11. The method of claim 9 wherein said polyurethane comprises aliphatic polyether.

12. The method of claim 11 wherein said aliphatic polyether polyurethane comprises about 45-100% water content.

13. The method of claim 1 wherein said outer membrane is layered directly onto and in contact with an enzyme layer.

14. The method of claim 13 wherein said enzyme layer comprises at least one enzyme that is stabilized in a matrix of the enzyme layer.

15. The method of claim 1 wherein the oxidoreductase enzyme is a creatinase.

16. The method of claim 1 wherein the oxidoreductase enzyme is a sarcosine oxidase.

6. A method for determining whether an elevated level of an analyte in a whole blood sample of a patient is an artifact related to hemolysis comprising:

introducing said whole blood sample to an electrochemical sensor, said electrochemical sensor comprising an oxidoreductase enzyme capable of generating hydrogen peroxide and a hydrophilic outer membrane comprising a thickness in a range of about 0.1 μ

m to about 50 μ

m, said membrane thickness adapted to enhance efflux of said hydrogen peroxide, followed by;

detecting an electrochemical signal generated by said hydrogen peroxide in the presence of hemoglobin (Hb(Fe²⁺)),

wherein a decrease of the electrochemical signal in a range of about 4% to about 50% as compared to a standard nor whole blood sample, is indicative of said hemolysis as the cause of the elevated level of the analyte in the whole blood sample of the patient.

10. The method of claim 6 wherein said outer membrane is composed of a polyurethane component.