[Determination of glycosylated hemoglobin by affinity chromatography on boron phenylagarose]

When Hb AIc was estimated by affinity chromatography where boron phenyl-agarose was used as a sorbent, the highest rate of Hb AIc binding with the sorbent was detected at pH 8.5-9.0 while the maximal elution of the protein occurred at 0.3-0.5 M concentration of sorbitol used as an eluent. Content of Hb AIc, estimated in 56 patients with insulin-dependent diabetes mellitus, constituted 12.3 +/- 0.4%, while in 20 healthy volunteers it was equal to 5.4 +/- 0.2%, which are consistent with the literature data obtained by other methods. At the same time, the data of Hb AIc estimation by means of affinity chromatography correlated exactly with the results of ion exchange chromatography (r = 0.98), thus corroborating the validity of the procedure used. Only the stable ketoamine fraction of Hb AIc was found to interact with boron phenyl-agarose.