Hileman R E, Siegel M M, Tabei K, Balagurunathan K, Linhardt R J
Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242, USA.
Electrophoresis. 1998 Nov;19(15):2677-81. doi: 10.1002/elps.1150191517.
A beta-cyclodextrin sulfate mixture has been fractionated using discontinuous gradient polyacrylamide gel electrophoresis. Semidry electrotransfer of the sample onto a positively charged nylon membrane and visualization of a portion of this membrane with Alcian blue stain showed multiple bands. The bands were cut from the remaining portion of the membrane 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 polyacrylamide gel electrophoresis as well as capillary electrophoresis, using indirect detection, showed some of the fractions to be pure while others were mixtures. Each beta-cyclodextrin sulfate fraction was complexed with a basic synthetic peptide and analyzed by electrospray ionization mass spectrometry to define the mass of the components in each mixture and thereby to determine the purity of each sample.
使用不连续梯度聚丙烯酰胺凝胶电泳对硫酸β-环糊精混合物进行了分级分离。将样品半干式电转移到带正电荷的尼龙膜上，并用阿尔辛蓝染色对该膜的一部分进行可视化，结果显示有多个条带。从膜的其余部分切下这些条带，用8 M尿素洗涤后，用2 M氯化钠洗脱硫酸β-环糊精级分并进行透析。使用高分辨率分析梯度聚丙烯酰胺凝胶电泳以及毛细管电泳对每个级分进行分析，采用间接检测法，结果显示一些级分为纯品，而其他级分为混合物。将每个硫酸β-环糊精级分与一种碱性合成肽复合，并通过电喷雾电离质谱法进行分析，以确定每种混合物中各组分的质量，从而确定每个样品的纯度。
