Determination of glycosylated hemoglobin by affinity electrophoresis on agarose gel.
We describe a method for electrophoretic determination of glycosylated hemoglobin (GHb) on agarose gel membrane. Clear separation of GHb from non-GHb is achieved after 30 min electrophoresis at 60 V using a sodium citrate buffer solution (34 mmol/l, pH 6.5) containing 0.6 g/l dextran sulfate. The results obtained by this method correlate well with those by agar gel electrophoresis (r = 0.98) and column chromatography (r = 0.96). However, unlike column chromatographic methods, GHb values obtained on agarose gel are not affected by fluctuations in ambient temperature (18-28 degrees C), changes in pH (6.2-6.6) and ionic strength of buffer solution (26-40 mmol/l), or variant hemoglobin S and C. Also, electrophoresis on agarose gel membrane eliminates the lengthy preparative steps required for cellulose acetate electrophoresis. We conclude that agarose gel electrophoresis is a simple method for quantitative determination of GHb.
D002846:Chromatography, Affinity / Q000379:methods; D016264:Dextran Sulfate; D003911:Dextrans; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006442:Glycated Hemoglobin / Q000032:analysis*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013696:Temperature
D016264:Dextran Sulfate; D003911:Dextrans; D006442:Glycated Hemoglobin