Glycosylated Hemoglobin Determinations by Isocratic High-Performance Liquid Chromatography

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Abstract
Glycosylated hemoglobins were separated isocratically by high-performance liquid chromatography (HPLC) on a cation-exchanger (CM-300). Both the stable and the labile fractions eluted together. The labile fraction was eliminated by incubating the red blood cells at 37[ddot]C for 20 min in a acidic buffer before injecting the sample on the column The column plate number was found to be dependent on the amount of sample injected. The capacity factor was dependent on the type of buffer, pH and ionic strength. Controls were preserved by preparing the hemolysates in 5% ethylene glycol. The method compared favorably with a commercial disposable minicolumn method.