Characterization of quantitation of glycosylated hemoglobins in canine blood

Canine hemolysates exhibited two minor and one major hemoglobins when chromatographed on Bio Rex-70 column with 0.05 M phosphate-0.01 M KCN (pH 6.5) containing 0.015 M NaCl. The first minor fraction was electrophoretically heterogeneous, constituting at least two hemoglobins. The second was completely homogeneous with somewhat faster anodal mobility than the major one and the difference was shown to be localized solely in the beta subunits. Thiobarbituric acid reaction was strongly positive for this hemoglobin, indicating the presence of glycosyl ketoamine linkage. We describe a simple and reliable procedure for quantitation of the canine glycosylated hemoglobins with a home-made Bio Rex-70 mini-column.