Baryla N E, Lucy C A
Department of Chemistry, University of Alberta, Edmonton, Canada.
Anal Chem. 2000 May 15;72(10):2280-4. doi: 10.1021/ac991191v.
The zwitterionic surfactant Rewoteric AM CAS U forms a dynamic wall coating that prevents the adsorption of cationic proteins as well as suppresses the electroosmotic flow (EOF). Addition of polarizable anions to buffers containing this zwitterionic surfactant increases the once suppressed EOF to values nearing +3 x 10(-4) cm2/(V s). The retention of the EOF allows for the separation of analytes of widely different mobilities and is demonstrated by the simultaneous separation of cationic and anionic proteins. Using a buffer containing optimal amounts of the polarizable anion perchlorate and surfactant CAS U, the proteins lysozyme, ribonuclease A, alpha-chymotrypsinogen A, and myoglobin are separated in less than 15 min. Efficiencies as high as 1.5 million plates/m and recoveries greater than 91% are observed for proteins injected in distilled water. Migration time reproducibility is approximately 1% RSD within 1 day and approximately 3% RSD from day to day. The anionic and cationic proteins can be separated over a pH range of 5.5-9, all yielding good efficiencies.
两性离子表面活性剂Rewoteric AM CAS U形成一种动态壁涂层，可防止阳离子蛋白吸附并抑制电渗流（EOF）。向含有这种两性离子表面活性剂的缓冲液中添加可极化阴离子，会使曾经被抑制的电渗流增加到接近+3×10⁻⁴ cm²/(V·s)的值。电渗流的保留使得迁移率差异很大的分析物得以分离，阳离子和阴离子蛋白的同时分离证明了这一点。使用含有最佳量可极化阴离子高氯酸盐和表面活性剂CAS U的缓冲液，溶菌酶、核糖核酸酶A、α-胰凝乳蛋白酶原A和肌红蛋白在不到15分钟内即可分离。对于注入蒸馏水中的蛋白质，观察到高达150万理论塔板数/米的柱效和大于91%的回收率。迁移时间重现性在1天内约为1%相对标准偏差，日复一日约为3%相对标准偏差。阴离子和阳离子蛋白可在pH 5.5 - 9的范围内分离，所有分离都具有良好的柱效。
