Hemoglobin sensor

• US 6,632,349 B1
• Filed: 07/14/2000
• Issued: 10/14/2003
• Est. Priority Date: 11/15/1996
• Status: Expired due to Term

• Current Assignee: Lifescan Incorporated (Duv Holding Corporation)
• Original Assignee: Lifescan Incorporated (Duv Holding Corporation)
• Inventors: Beck, Thomas, Hodges, Alastair, Chatelier, Ron
• Primary Examiner(s): Nguyen, Nam
• Assistant Examiner(s): Olsen, Kaj K.
• Application Number:
  US09/616,512
• Time in Patent Office: 1,187 Days
• Field of Search: 204/400, 204/403, 204/409, 204/403.01, 204/403.02, 205/777.5, 205/778, 205/792, 156/292, 435/7.4, 435/817
• US Class Current: 205/792
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH

Abstract

The present invention relates to a device and method for measuring hemoglobin in a fluid sample. The device comprises a disposable electrochemical cell, such as a thin layer electrochemical cell, containing a reagent capable of being reduced by hemoglobin. A suitable fluid sample that may be analyzed according the present invention is whole blood. If the hemoglobin to be analyzed is present in red blood cells, a lysing agent may be added to the sample to release the hemoglobin prior to analysis.

Claims

1. A device for measuring an amount of hemoglobin in a fluid whole blood sample, the whole blood sample containing red blood cells, the red blood cells containing hemoglobin, the device comprising an electrochemical cell having a sensing chamber, a first electrode, a second electrode wherein the second electrode is mounted in opposing relationship a distance of less than about 500 microns from the first electrode, an aperture for admitting the sample into the sensing chamber, a red blood cell lysing agent, and a reagent contained within the sensing chamber wherein the reagent is capable of being reduced by hemoglobin to generate an electrical signal indicative of the amount of hemoglobin, wherein the device further comprises means for applying a positive potential to the first electrode;

means for measuring a current generated by reaction of the reagent and hemoglobin;

means for analyzing the current to give a result, the result comprising a time required for substantial lysis of the red blood cells or a derived final value for the current;

means for calculating a percentage of the reaction completed as a function of time based on the result of the analyzing step;

means for reversing the potential on the first electrode;

means for measuring a transient current; and

means for determining a diffusion coefficient and a concentration of a reduced form of the reagent based on the transient current.

2. The device of claim 1, wherein the device may only be used for a single test.

3. The device of claim 1, wherein the first electrode comprises a sensing electrode.

4. The device of claim 1, wherein the first electrode comprises a material selected from the group consisting of platinum, palladium, carbon, indium oxide, tin oxide, gold, iridium, copper, steel, silver, and mixtures thereof.

5. The device of claim 1, wherein the first electrode is formed by a technique selected from the group consisting of sputtering, vapor coating, screen printing, thermal evaporation, ink jet printing, ultrasonic spraying, slot coating, gravure printing and lithography.

6. The device of claim 1, wherein the second electrode comprises a counter electrode.

7. The device of claim 1, wherein the second electrode comprises a metal in contact with a metal salt.

8. The device of claim 7, wherein the metal in contact with a metal salt is selected from the group consisting of silver in contact with silver chloride, silver in contact with silver bromide, silver in contact with silver iodide, mercury in contact with mercurous chloride, and mercury in contact with mercurous sulfate.

9. The device of claim 7, the electrochemical cell further comprising a third electrode.

10. The device of claim 9, wherein the third electrode comprises a reference electrode.

11. The device of claim 10, wherein the third electrode comprises a metal in contact with a metal salt.

12. The device of claim 11, wherein the metal in contact with the metal salt is selected from the group consisting of silver in contact with silver chloride, silver in contact with silver bromide, silver in contact with silver iodide, mercury in contact with mercurous chloride, and mercury in contact with mercurous sulfate.

13. The device of claim 1, wherein the second electrode comprises a reference electrode.

14. The device of claim 1, wherein the reagent is selected from the group consisting of dichromate, vanadium oxides, permanganate, electroactive organometallic complexes, quinones, and dichlorophenolindophenol.

15. The device of claim 1, wherein the reagent comprises ferricyanide.

16. The device of claim 1, the sensing chamber further comprising a buffer, wherein the buffer is contained within the sensing chamber.

17. The device of claim 16, wherein the buffer is selected from the group consisting of phosphates, carbonates, alkali metal salts of mellitic acid, and alkali metal salts of citric acid.

18. The device of claim 1, wherein the lysing agent is selected from the group consisting of ionic detergents, nonionic detergents, proteolytic enzymes, and lipases.

19. The device of claim 1, wherein the lysing agent comprises saponin.

20. The device of claim 1, wherein the lysing agent is selected from the group consisting of sodium dodecyl sulfate, cetyl trimethylammonium bromide, and polyethoxylated octylphenol.

21. The device of claim 1, the sensing chamber further comprising a support, the support contained within the sensing chamber.

22. The device of claim 21, wherein the support is a material selected from the group consisting of mesh, nonwoven sheet, fibrous filler, macroporous membrane, sintered powder, and combinations thereof.

23. The device of claim 21, wherein the reagent is contained within or supported on the support.

24. The device of claim 21, further comprising a buffer, wherein the buffer is contained within or supported on the support.

25. The device of claim 21, wherein the lysing agent is contained within or supported on the support.

26. The device of claim 1, wherein the second electrode is mounted in opposing relationship a distance of less than about 150 microns from the first electrode.

27. The device according to claim 1, wherein the second electrode is mounted in opposing relationship a distance of less than about 150 microns and greater than about 50 microns from the first electrode.

28. The device of claim 1, further comprising an interface for communication with a meter.

29. The device of claim 28, wherein the interface communicates a voltage or a current.

30. The device of claim 1, wherein the electrochemical cell comprises a thin layer electrochemical cell.

31. A method for measuring an amount of hemoglobin in a fluid whole blood sample, the whole blood sample containing red blood cells, the red blood cells containing hemoglobin, wherein the method comprises:

providing a device comprising an electrochemical cell having a sensing chamber, a first electrode, a second electrode wherein the second electrode is mounted in opposing relationship a distance of less than about 500 microns from the first electrode, an aperture for admitting the sample into the sensing chamber;

a reagent contained within the sensing chamber and capable of being reduced by hemoglobin; and

a red blood cell lysing agent contained within the sensing chamber;

placing the whole blood sample in the sensing chamber, whereby the lysing agent contained within the sensing chamber releases hemoglobin from the red blood cells, whereby the hemoglobin thus released reduces the reagent; and

obtaining an electrochemical measurement indicative of an amount of hemoglobin present in the whole blood sample, wherein the step of obtaining an electrochemical measurement comprises;

applying a positive potential to the first electrode;

measuring a current generated by reaction of the reagent and hemoglobin;

analyzing the current to give a result, the result comprising a time required for substantial lysis of the red blood cells or a derived final value for the current;

calculating a percentage of the reaction completed as a function of time based on the result of the analyzing step;

reversing the potential on the first electrode;

measuring a transient current; and

determining a diffusion coefficient and a concentration of a reduced form of the reagent based on the transient current.

32. The method of claim 31, wherein the device may only be used for a single test.

33. The method of claim 31, wherein the electrochemical cell is a thin layer electrochemical cell.

34. The method of claim 31, wherein the first electrode comprises a sensing electrode, the sensing electrode comprising a material selected from the group consisting of platinum, palladium, carbon, indium oxide, tin oxide, gold, iridium, copper, steel, silver, and mixtures thereof.

35. The method of claim 31, wherein the second electrode comprises a counter electrode, the counter electrode comprising a metal in contact with a metal salt.

36. The method of claim 35, wherein the metal in contact with a metal salt is selected from the group consisting of silver in contact with silver chloride, silver in contact with silver bromide, silver in contact with silver iodide, mercury in contact with mercurous chloride, and mercury in contact with mercurous sulfate.

37. The method of claim 31, the electrochemical cell further comprising a third electrode, the third electrode comprising a reference electrode.

38. The method of claim 31, wherein the reagent is selected from the group consisting of dichromate, vanadium oxides, permanganate, electroactive organometallic complexes, quinones, dichlorophenolindophenol, and ferricyanide.

39. The method of claim 31, the sensing chamber further comprising a buffer, wherein the buffer is contained within the sensing chamber.

40. The method of claim 39, wherein the buffer is selected from the group consisting of phosphates, carbonates, alkali metal salts of mellitic acid, and alkali metal salts of citric acid.

41. The method of claim 31, wherein the lysing agent is selected from the group consisting of ionic detergents, nonionic detergents, proteolytic enzymes, lipases, saponin, sodium dodecyl sulfate, cetyl trimethylanmmonium bromide, and polyethoxylated octylphenol.

42. The method of claim 31, the sensing chamber further comprising a support, the support contained within the sensing chamber.

43. The method of claim 31, wherein the second electrode is mounted in opposing relationship a distance of less than about 150 microns from the first electrode.

44. The method of claim 31, wherein the second electrode is mounted in opposing relationship a distance of less than about 150 microns and greater than about 50 microns from the first electrode.

45. The method of claim 31, wherein the electrochemical cell comprises a thin layer electrochemical cell.

46. A method of manufacture of a device for determining an amount of hemoglobin in a fluid whole blood sample, the whole blood sample comprising red blood cells, the red blood cells comprising hemoglobin, the device comprising an electrochemical cell having a sensing chamber, a first electrode, a second electrode, an aperture for admitting the sample into the sensing chamber, a red blood cell lysing agent, and a reagent contained within the sensing chamber, wherein the reagent is capable of being reduced by hemoglobin to generate an electrical signal indicative of the amount of hemoglobin in the sample, the method comprising:

forming an aperture extending through a sheet of electrically resistive material, the aperture defining a side wall of the sensing chamber;

mounting a first layer having a first electrode to a first side of the sheet and extending over the aperture whereby to define a first sensing chamber end wall, the first electrode facing the first side of the sheet;

mounting a second layer having a second electrode to a second side of the sheet and extending over the aperture whereby to define a second sensing chamber end wall in substantial overlying registration with the first layer, the second electrode facing the second side of the sheet, whereby the sheet and layers form a strip, and wherein the second electrode is mounted in opposing relationship a distance of less than about 500 microns from the first electrode;

providing an aperture in the strip to permit entry of sample into the sensing chamber;

providing a reagent capable of being reduced by hemoglobin, wherein the reagent is contained within the sensing chamber;

providing a red blood cell lysing agent, wherein the lysing agent is contained within the sensing chamber;

providing means for applying a positive potential to the first electrode;

means for measuring a current generated by reaction of the reagent and hemoglobin;

providing means for analyzing the current to give a result, the result comprising a time required for substantial lysis of red blood cells or a derived final value for the current;

providing means for calculating a percentage of the reaction completed as a function of time based on the result of the analyzing step;

providing means for reversing the potential on the first electrode;

providing means for measuring a transient current; and

providing means for determining a diffusion coefficient and a concentration of a reduced form of the reagent based on the transient current.

47. The method according to claim 46, further comprising the step of providing a vent in the strip to permit the escape of air displaced from the sensing chamber as sample fills the sensing chamber.

48. The method according to claim 46, wherein at least one of the electrodes comprises a noble metal.

49. The method according to claim 48, wherein the noble metal is selected from the group consisting of palladium, platinum, and silver.

50. The method according to claim 46, wherein the aperture is of a rectangular cross-section.

51. The method according to claim 46, wherein at least one of the electrodes is a sputter coated metal deposit.

52. The method according to claim 46, wherein the electrodes are adhered to the sheet.

53. The method according to claim 52, wherein the electrodes are adhered to the sheet by an adhesive.

54. The method according to claim 53, wherein the adhesive is selected from the group consisting of heat activated adhesive, pressure sensitive adhesive, heat cured adhesive, chemically cured adhesive, hot melt adhesive, and hot flow adhesive.

55. The method according to claim 46, further comprising the step of providing a buffer, the buffer being contained within the sensing chamber.

56. The method according to claim 55, further comprising the step of printing the buffer onto at least one wall of the sensing chamber.

57. The method according to claim 46, further comprising the step of printing the red blood cell lysing agent onto at least one wall of the sensing chamber.

58. The method according to claim 46, further comprising the step of printing the reagent onto at least one wall of the sensing chamber.

59. The method according to claim 46, further comprising the step of providing a support, the support contained within the sensing chamber.

60. The method according to claim 59, wherein the support is selected from the group consisting of mesh, fibrous filler, macroporous membrane, sintered powder, and combinations thereof.

61. The method according to claim 60, wherein the reagent is supported on or contained within the support.

62. The method according to claim 46, wherein at least the sheet or one of the layers comprises a polymeric material selected from the group consisting of polyester, polystyrene, polycarbonate, polyolefin, and mixtures thereof.

63. The method according to claim 46, wherein at least the sheet or one of the layers comprises polyethylene terephthalate.

64. The method according to claim 46, wherein the second electrode is mounted in opposing relationship a distance of less than about 150 microns from the first electrode.

65. The method according to claim 46, wherein the second electrode is mounted in opposing relationship a distance of less than about 150 microns and greater than about 50 microns from the first electrode.

66. The method according to claim 46, wherein the device may only be used for a single-test.