A comparison of three methods for quantitation of variant hemoglobin fractions

Three methods, cellulose acetate electrophoresis-densitometry (CAE-D), cellulose acetate electrophoresis-elution (CAE-E), and microchromatography (MGC), were evaluated to determine the best method for quantitating variant hemoglobins. Evaluations were performed in two phases. In Phase I samples of known hemoglobins were obtained and Hb S and/or Hb C were quantitated by the three methods to evaluate agreement of results. In Phase II reference samples were prepared. The Hb S concentrations from each method were compared to the reference value to determine the effect of varying concentrations of Hb S on the methods. Cellulose acetate electrophoresis-densitometry proved to be the best method for quantitating variant hemoglobins in which the concentration was greater than 35 percent. Microchromatography was shown to be the best method when Hb S was approximately 30 percent. Either the microchromatography or the cellulose acetate electrophoresis-elution method proved to be a good method for quantitating approximately 25 percent Hb S.