ISOLATION AND CHARACTERIZATION OF BETA-CYCLODEXTRIN SULFATES BY PREPARATIVE GRADIENT POLYACRYLAMIDE-GEL ELECTROPHORESIS, CAPILLARY-ELECTROPHORESIS AND ELECTROSPRAY IONIZATION-MASS SPECTROMETRY

A beta-cyclodextrin sulfate mixture has been fractionated using discon tinuous gradient polyacrylamide gel electrophoresis. Semidry electrotr ansfer of the sample onto a positively charged nylon membrane and visu alization of a portion of this membrane with Alcian blue stain showed multiple bands. The bands were cut from the remaining portion of the m embrane and after washing with 8 M urea, the beta-cyclodextrin sulfate fractions were eluted with 2 M sodium chloride and dialyzed. Analysis of each fraction using high resolution analytical gradient polyacryla mide gel electrophoresis as well as capillary electrophoresis, using i ndirect detection, showed some of the fractions to be pure while other s were mixtures. Each beta-cyclodextrin sulfate fraction was complexed with a basic synthetic peptide and analyzed by electrospray ionizatio n mass spectrometry to define the mass of the components in each mixtu re and thereby to determine the purity of each sample.