Methods and reagents for cyanide-free determination of hemoglobin and leukocytes in whole blood

  • US 5,958,781 A
  • Filed: 05/27/1997
  • Issued: 09/28/1999
  • Est. Priority Date: 07/14/1994
  • Status: Expired due to Term
First Claim
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1. A method for making hemoglobin (Hb) and white blood cell (WBC) determinations comprising, in combination:

  • (a) diluting a whole blood sample with a diluent to give a diluted whole blood test sample, said diluent consisting of;

    sodium sulfate, sodium chloride, EDTA, imidazole, and a mixture of hexahydro-1,3,5-tris (2-hydroxyethyl)-s-triazine and sodium 2-pyridinethiol-1-oxide;

    (b) mixing said diluted whole blood test sample with an aqueous multipurpose lyse reagent to give a whole blood sample/lyse reagent mixture, said whole blood sample/lyse reagent mixture forming a chromogen, said multipurpose lyse reagent comprising from about 0.1 to about 20% by weight of at least one quaternary ammonium salt, said at least one quaternary ammonium salt selected from the group consisting of;

    tetradecyltrimethyl ammonium bromide (TTAB), dodecyltrimethyl ammonium chloride, cetyltrimethyl ammonium bromide, hexadecyltrimethy ammonium bromide, benzalkonium chloride, and cetylpyridium chloride and from about 0.1 to about 15% by weight of at least one hydroxylamine salt selected from the group consisting of;

    hydrochloride, sulfate, phosphate and other acid salts;

    (c) presenting said whole blood sample/lyse reagent mixture to an absorbance spectrophotometer;

    (d) measuring the optical density of the chromogen;

    (e) determining total Hb concentration in the sample from the measured optical density of the chromogen, wherein the chromogen has a maximum absorbance of from about 530 nm to about 550 nm;

    (f) presenting said whole blood sample/lyse reagent mixture to a particle analyzing system;

    (g) detecting WBC populations; and

    (h) determining the number of white blood cells in said WBC populations in said whole blood sample.

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