Separation of human globin chains by micellar electrokinetic capillary chromatography

A new separation method of human globin chains by micellar electrokinetic c apillary chromatography (MECC) is described. In this method, a 25 mM phosph ate buffer (pH 2.5) containing 7 M urea and 1% (w/v) reduced Triton X-100 b uffer system was used. All experiments were performed in a 47 cmX50 mu m I. D. uncoated fused-silica capillary. The separation voltage was set at 19 kV . Normal globin chains derived from normal adults and newborns, alpha, beta , delta, (G)gamma and (A)gamma globin chains as well as common variant glob in chains were successfully separated within 20 min. High reproducible migr ation times of globin chains (CVs of intra- and inter-assay were less than 1% and 2% respectively), and quantification of (G)gamma and (A)gamma chains (CVs for intra- and inter-assay were less than 5% and 10%, respectively) w ere obtained. This new MECC method provides primary information on structur al modification of globin chains. It can be an important diagnostic tool in clinical laboratory practice in the field of hemoglobinopathies. (C) 1998 Elsevier Science B.V. All rights reserved.