SEPARATION OF HEMOGLOBIN-VARIANTS IN SINGLE HUMAN ERYTHROCYTES BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED NATIVE FLUORESCENCE DETECTION

Single human red blood cells, in which the hemoglobin (Hb) molecules e xist in their native, tetrameric states, were analyzed. Upon injection and lysis of a cell, the tetramers were dissociated on-column into th eir respective polypeptide chains, separated, and detected by laser-in duced native fluorescence detection with 275-nm excitation. This techn ique was applied to the determination of hemoglobin variants as found in adult (normal and elevated Hb A(1)) and fetal erythrocytes. Normal adult cells contained 9.6% and 4.8% glycated beta- and alpha-chains, r espectively. Cells with elevated Hb A(1) gave 30% and 12%, respectivel y. The amounts of glycated Hb and total Hb in a given cell were found to be uncorrelated.