Moritani Tohei, Yoon Kyunghwan, Rafailovich Miriam, Chu Benjamin
Department of Chemistry, State University of New York at Stony Brook, Stony Brook, NY 11794-3400, USA.
Electrophoresis. 2003 Aug;24(16):2764-71. doi: 10.1002/elps.200305546.
Two new methods of inner capillary coating with poly(vinyl alcohol) (PVAL) have been investigated and evaluated by performing DNA capillary electrophoresis (CE) using PVAL as a separation medium and by measuring the electroosmotic flow (EOF) mobility. The treatment of capillaries with a silanol-group modified PVAL (PVAL-Si) has been found to give good coating effects for improving the resolution of DNA CE and for reducing the EOF. This coating must be effectively achieved by combining the adsorptive property of PVAL chains onto silica with the reaction between the silanol groups of PVAL-Si and the silica surface. The adsorption of PVAL onto silica has been observed by using atomic force microscopy (AFM) for PVAL-Si as well as for a nonmodified PVAL as a control. The coating with PVAL that links to the capillary wall surface with more hydrolytically stable bonding, -Si-C-, has been formed by performing the Grignard reaction, followed by in-capillary polymerization of vinyl acetate (VAc) and hydrolysis. This coating has been found to be effective for improving the resolution of DNA CE and for reducing the EOF.
研究并评估了两种用聚乙烯醇(PVAL)进行毛细管内涂层的新方法,通过以PVAL作为分离介质进行DNA毛细管电泳(CE)以及测量电渗流(EOF)迁移率来实现。已发现用硅醇基修饰的PVAL(PVAL-Si)处理毛细管,对于提高DNA CE的分辨率和降低EOF具有良好的涂层效果。这种涂层必须通过将PVAL链在二氧化硅上的吸附特性与PVAL-Si的硅醇基团和二氧化硅表面之间的反应相结合才能有效实现。通过使用原子力显微镜(AFM)观察了PVAL-Si以及作为对照的未修饰PVAL在二氧化硅上的吸附情况。通过进行格氏反应,随后进行醋酸乙烯酯(VAc)的毛细管内聚合和水解,形成了以水解稳定性更高的键合-Si-C-与毛细管管壁表面相连的PVAL涂层。已发现这种涂层对于提高DNA CE的分辨率和降低EOF是有效的。