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Method and kit for chromatographic separation of hemoglobin A.sub.1c

  • US 4,389,491 A
  • Filed: 04/26/1982
  • Issued: 06/21/1983
  • Est. Priority Date: 04/26/1982
  • Status: Expired due to Term
First Claim
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1. A method for the separation of hemoglobin A1c from other glycosylated and nonglycosylated hemoglobins and the Schiff base precursor to hemoglobin A1c in a sample of human blood which comprises:

  • (a) lysing the red blood cells contained in said sample to form a hemolysate containing said hemoglobin A1c, said glycosylated and nonglycosylated hemoglobins, and said Schiff base precursor,(b) impregnating a weak cation exchanger with said hemolysate,(c) passing through said exchanger a first buffer solution with ions of an alkali metal dissolved therein at a concentration of from about 0.02 M to about 0.05 M to dissociate said Schiff base precursor into glucose and hemoglobin A and to preferentially elute said glucose and said other glycosylated hemoglobins over said hemoglobin A, said hemoglobin A1c and said other nonglycosylated hemoglobins,(d) passing through said exchanger a second buffer solution containing ions of an alkali metal dissolved therein at a concentration of from about 0.06 M to about 0.11 M to preferentially elute said hemoglobin A1c over said hemoglobin A and said other nonglycosylated hemoglobins, and(e) recovering the eluate from step (d).

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