| abstract |
A cyanide-free lytic reagent composition and method for measuring the total hemoglobin concentration in a blood sample, for counting the number of leukocytes and for deferential counting of three leukocyte subpopulations including lymphocytes, monocytes, and granulocytes are described. The cyanide-free lytic reagent composition includes a hemolytic surfactant chosen from quaternary ammonium salts, pyridinium salts, organic phosphate esters, and alkyl sulfonates to lyse erythrocytes and release hemoglobin, and an organic ligand chosen from triazole and its derivatives, tetrazole and its derivatives, alkaline metal salts of oxonic acid, melamine, aniline-2-sulfonic acid, quinaldic acid, 2-amino-1,3,4-thiadiazole, triazine and its derivatives, urazole, DL-pipecolinic acid. isonicotinamide, anthranilonitrile, 6-aza-2-thiothymine, adenine, 3-(2-thienyl)acrylic acid, benzoic acid and alkali metal and ammonium salts of benzoic acid, and pyrazine and its derivatives to form a stable chromogen with hemoglobin. The lytic reagent composition has a pH ranging from about 1 to about 13. The lytic reagent composition is mixed with a blood sample which is prediluted with a suitable blood diluent and the UV absorption of the sample mixture is measured at the predetermined absorption wavelength of the formed chromogen from 510 nm to 560 nm. Counting the numbers of leukocytes and differential counting of three leukocyte subpopulations are accomplished simultaneously on an automated cell counter utilizing DC impedance measurement. Alternatively, the organic ligands can be added to a suitable blood diluent for hemoglobin and leukocyte analysis. |