Analysis method and cuvette therefor

• US 6,638,769 B2
• Filed: 10/10/2001
• Issued: 10/28/2003
• Est. Priority Date: 06/28/2000
• Status: Expired due to Term

• Current Assignee: Migrata U.K. Limited (Desert Shield Technologies Ltd.)
• Original Assignee: Migrata U.K. Limited (Desert Shield Technologies Ltd.)
• Inventors: Eriksson, Annika, Nilsson, Sven-Erik, Lilja, Jan, Svensson, Johnny
• Primary Examiner(s): WALLENHORST, MAUREEN
• Application Number:
  US09/972,918
• Publication Number:
  US 20020058342A1
• Time in Patent Office: 748 Days
• Field of Search: 436/63, 436/66, 436/164, 436/165, 436/166, 422/58, 422/82.05, 422/82.09, 422/99, 422/102, 422/939, 422/940
• US Class Current: 436/66
• CPC Class Codes: G01N 2021/0321 One time use cells, e.g. integrally moulded
• CPC Class Codes: G01N 2021/0325 Cells for testing reactions, e.g. containing reagents
• CPC Class Codes: G01N 2021/0346 Capillary cells; Microcells
• CPC Class Codes: G01N 2021/0389 Windows
• CPC Class Codes: G01N 21/03 Cuvette constructions
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 21/272 for following a reaction, e.g. for determining photometrically a reaction rate (photometric cinetic analysis)
• CPC Class Codes: G01N 21/31 Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry G01N21/72 takes precedence
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/72 involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood

Abstract

A method for hemoglobin determination is described which includes the steps of introducing a sample of undiluted whole blood by capillary action into a disposable microcuvette having at least one cavity for receiving the sample. The cavity includes a dry essentially non-hygroscopic hemolysing agent, which is dissolved by the blood, hemolyses the red blood cells and releases the haemoglobin contained in the blood cells. A first absorption measurement at a wavelength range 490-520 nm is then performed directly on the sample in the microcuvette, and a second absorption measurement is performed to compensate for background interference.

Claims

1. A method for quantitative haemoglobin determination in undiluted whole blood comprising the steps of:

introducing a sample of undiluted whole blood containing red blood cells having haemoglobin therein by capillary action into a disposable microcuvette having at least one cavity for receiving the sample, the cavity including an essentially non-hygroscopic hemolysing agent in a dry form, the cavity being essentially free of azide and nitrite, whereby the hemolysing agent is dissolved in the whole blood, the agent hemolyses the red blood cells and the haemoglobin contained in the blood cells is released;

performing a first absorption measurement at a wavelength range directly on the hemolysed sample in the cuvette,

further conducting a second absorption measurement on the hemolysed sample to compensate for background interference, and

analyzing the results of the absorption measurements and determining the concentration of haemoglobin in the whole blood sample.

2. The method according to claim 1 wherein the hemolysing agent is soluble in organic solvents.

3. The method according to claim 2 wherein the organic solvent is an alcohol.

4. The method according to claim 3 wherein the hemolysing agent present in the microcuvette permits a rapid introduction of whole blood into the cuvette.

5. The method according to claim 2 wherein the hemolysing agent U present in the microcuvette, permits a rapid introduction of whole blood into the cuvette.

6. The method according to claim 1 wherein the hemolysing agent present in the microcuvette, permits a rapid introduction of whole blood into the cuvette.

7. The method according to claim 6 wherein the time for the introduction of whole blood into the microcuvette is shorter than the time required for dissolving the hemolysing agent.

8. The method according to claim 1 wherein the hemolysing agent is selected from the group consisting of ionic and nonionic, surface active substances.

9. The method according to claim 1 wherein the hemolysing agent is selected from the group consisting of salts of deoxycholic acid and quaternary ammonium salts.

10. The method according to claim 1 wherein the hemolysing agent is selected from the group consisting of sodium deoxycholate, potassium deoxycholate, calcium deoxychoholate, morfolin deoxycholate, cyclohexylammonium deoxycholate, ammonium deoxycholate and mixtures thereof.

11. The method according to claim 1 wherein the hemolysing agent essentially consists of a mixture of sodium deoxycholate and ammonium deoxycholate.

12. The method according to claim 11 wherein the amount of ammonium deoxycholate is between 20 and 80 percent by weight.

13. The method according to claim 1 wherein the second absorption measurement is performed in the range of 850-910 nm.

14. The method according to claim 1 wherein the first absorption measurement is performed in the range of 500-510 nm.

15. The method according to claim 1 wherein the second absorption measurement is performed in the range of 860-900 nm.

16. Disposable microcuvette for spectrophotometric determination of haemoglobin in undiluted whole blood, wherein the microcuvette comprises a cavity containing a dried, non-hygroscopic hemolysing agent or a combination of said agents, provided that the cavity is essentially free from azide and nitrite, wherein the hemolysing agent is a salt of deoxycholic acid.

17. The disposable microcuvette according to claim 16, wherein the hemolysing agent is selected from the group consisting of sodium deoxycholate, potassium deoxycholate, calcium deoxycholate, morfolin deoxycholate, cyclohexylammonium deoxycholate, ammonium deoxycholate and mixtures thereof.

18. The disposable microcuvette according to claim 16, wherein the hemolysing agent essentially consists of a mixture of sodium deoxycholate and ammonium deoxycholate.

19. The disposable microcuvette according to claim 18, wherein the amount of ammonium deoxycholate is between 20 and 80 percent by weight of the mixture.