Claims
- 1. A method for the determination, quantification and monitoring of extracellular hemoglobin in a whole blood, plasma or serum sample performed on an automated hematology analyzer having at least two analytic channels for hemoglobin measurement, comprising:
(a) determining total hemoglobin concentration in an aliquot of the sample in a hemoglobin analytic channel of the analyzer; (b) determining cellular hemoglobin concentration in the sample aliquot in a red blood cell analytic channel of the analyzer; and (c) calculating the difference between the total hemoglobin concentration determined from the hemoglobin channel and the cellular hemoglobin concentration determined from the red blood cell channel of the analyzer to yield the extracellular hemoglobin concentration in the whole blood, plasma, or serum sample.
- 2. The method according to claim 1, wherein the sample is a normal whole blood sample.
- 3. The method according to claim 1, wherein the sample is an abnormal whole blood sample.
- 4. The method according to claim 1, wherein the sample is a plasma or serum sample.
- 5. The method according to claim 1, wherein the total hemoglobin concentration in step (a) is from about 0.5 g/dL to about 25 g/dL of blood, plasma, or serum.
- 6. The method according to claim 5, wherein the total hemoglobin concentration in step (a) is from about 6 g/dL to about 22 g/dL of blood, plasma, or serum.
- 7. The method according to claim 1, wherein the cellular hemoglobin concentration in step (b) is from about 0.5 g/dL to about 25 g/dL of blood, plasma, or serum.
- 8. The method according to claim 7, wherein the cellular hemoglobin concentration in step (b) is from about 4 g/dL to about 24 g/dL of blood, plasma, or serum.
- 9. The method according to claim 8, wherein the cellular hemoglobin concentration is about 6.0 g/dL of blood, plasma, or serum.
- 10. The method according to claim 3, wherein the abnormal whole blood sample is derived from an individual having a pathological condition.
- 11. The method according to claim 10, wherein the pathological condition is selected from blood loss during surgery, blood loss during trauma, and hemorrhagic shock.
- 12. The method according to claim 1, wherein the hemoglobin analytic channel of step a) measures the total hemoglobin concentration in the sample by hemolysis and extraction of the hemes from their biological complex with globin by forming a ligated ferric heme species capturable in a surfactant micelle.
- 13. The method according to claim 1, wherein the red blood cell analytic channel of step b) measures red cell concentration, red cell volume, and hemoglobin concentration of individual red blood cells as they pass through two light scattering detectors virtually one cell at a time.
- 14. The method according to claim 1, wherein the total hemoglobin concentration of step (a) is determined from calorimetric absorbance in the hemoglobin channel of the analyzer.
- 15. The method according to claim 1, wherein the cellular hemoglobin of step (b) is determined by the formula:
- 16. The method according to claim 1, wherein the extracellular hemoglobin in the sample is due to the presence of a cell-free hemoglobin product or an oxygen-carrying hemoglobin substitute in the blood.
- 17. The method according to claim 16, wherein the cell-free hemoglobin product or the oxygen-carrying hemoglobin substitute is selected from the group consisting of purified hemoglobin, recombinant hemoglobin, cross-linked hemoglobin, polymerized hemoglobin and hemoglobin coupled to polyethylene glycol (PEG-HGB).
- 18. The method according to claim 17, wherein the purified hemoglobin is purified bovine hemoglobin or purified human hemoglobin.
- 19. The method according to claim 17, wherein the recombinant hemoglobin is recombinant human hemoglobin.
- 20. An automated method for the determination, quantification and monitoring of extracellular hemoglobin in a whole blood, plasma or serum sample performed on an automated hematology analyzer having at least two analytic channels for hemoglobin measurement, comprising:
(a) determining total hemoglobin concentration in an aliquot of the whole blood, plasma, or serum sample in a hemoglobin analytic channel of the analyzer, wherein said hemoglobin analytic channel measures the total hemoglobin concentration in the sample by hemolysis and extraction of hemes from their biological complex with globin by forming a ligated ferric heme species capturable in a surfactant micelle, and further wherein the total hemoglobin concentration is determined from calorimetric absorbance in the hemoglobin channel of the analyzer; (b) determining cellular hemoglobin concentration in the sample aliquot in a red blood cell analytic channel of the analyzer, wherein the red blood cell analytic channel measures red cell concentration, red cell volume and hemoglobin concentration of individual red blood cells as they pass through two light scattering detectors virtually one cell at a time; and (c) calculating the difference between the total hemoglobin concentration determined from the hemoglobin channel and the cellular hemoglobin concentration determined from the red blood cell channel of the analyzer to yield the extracellular hemoglobin concentration in the whole blood, plasma, or serum sample.
- 21. The method according to claim 20, wherein the sample is a normal whole blood sample or an abnormal whole blood sample.
- 22. The method according to claim 20, wherein the sample is a plasma or serum sample.
- 23. The method according to claim 20, wherein the total hemoglobin concentration in step (a) is from about 0.5 g/dL to about 25 g/dL of blood, plasma, or serum; and the cellular hemoglobin concentration in step (b) is from about 0.5 g/dL to about 25 g/dL of blood, plasma, or serum.
- 24. The method according to claim 23, wherein the cellular hemoglobin concentration is about 6.0 g/dL.
- 25. The method according to claim 20, wherein the abnormal blood sample is derived from an individual having a pathological condition.
- 26. The method according to claim 25, wherein the pathological condition is selected from the group consisting of blood loss during surgery, blood loss during trauma, and hemorrhagic shock.
- 27. The method according to claim 20, wherein the extracellular hemoglobin in the sample is due to the presence of a cell-free hemoglobin product or an oxygen-carrying hemoglobin substitute in the blood.
- 28. The method according to claim 27, wherein the cell-free hemoglobin product or the oxygen-carrying hemoglobin substitute is selected from purified hemoglobin, recombinant hemoglobin, cross-linked hemoglobin, polymerized hemoglobin, purified bovine hemoglobin and hemoglobin coupled to polyethylene glycol (PEG-HGB).
- 29. The method according to claim 28, wherein the purified hemoglobin is purified bovine hemoglobin or purified human hemoglobin.
- 30. The method according to claim 28, wherein the recombinant hemoglobin is recombinant human hemoglobin.
- 31. The method according to claim 20, wherein the cellular hemoglobin of step (b) is determined by the formula:
Parent Case Info
[0001] This application claims benefit of patent application U.S. Ser. No. 60/210,625, filed Jun. 9, 2000.
Provisional Applications (1)
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Number |
Date |
Country |
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60210625 |
Jun 2000 |
US |