Bendahl L, Hansen S H, Gammelgaard B
Department of Analytical and Pharmaceutical Chemistry, Royal Danish School of Pharmacy, Copenhagen, Denmark.
Electrophoresis. 2001 Aug;22(12):2565-73. doi: 10.1002/1522-2683(200107)22:12<2565::AID-ELPS2565>3.0.CO;2-I.
A simple coating procedure for generation of a high and pH-independent electroosmotic flow in capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MEKC) is described. The bilayer coating was formed by noncovalent adsorption of the ionic polymers Polybrene and poly(vinylsulfonate) (PVS). A stable dynamic coating was formed when PVS was added to the background electrolyte. Thus, when the PVS concentration in the background electrolyte was optimized for CZE (0.01%), the EOF differed less than 0.3% after 54 runs. The electroosmotic mobility in the coated capillaries was (4.9+/-0.1) x 10(-4) cm2V(-1)s(-1) in a pH-range of 2-10 (ionic strength = 30 mM). When alkaline compounds were used as test substances intracapillary and intercapillary migration time variations (n = 6) were less than 1% relative standard deviation (RSD) and 2% RSD, respectively in the entire pH range. The coating was fairly stable in the presence of sodium dodecyl sulfate, and this made it possible to perform fast MEKC separations at low pH. When neutral compounds were used as test substances, the intracapillary migration time variations (n = 6) were less than 2% RSD in a pH range of 2-9. In addition to fast CZE and MEKC separations at low pH, analysis of the alkaline compounds by CE-MS was also possible.
本文描述了一种在毛细管区带电泳(CZE)和胶束电动毛细管色谱(MEKC)中产生高且与pH无关的电渗流的简单涂层方法。双层涂层由离子聚合物聚凝胺和聚乙烯磺酸盐(PVS)通过非共价吸附形成。当将PVS添加到背景电解质中时,形成了稳定的动态涂层。因此,当针对CZE将背景电解质中的PVS浓度优化为0.01%时,在54次运行后电渗流的差异小于0.3%。在pH值为2至10(离子强度 = 30 mM)的范围内,涂覆毛细管中的电渗迁移率为(4.9±0.1)×10⁻⁴ cm²V⁻¹s⁻¹。当使用碱性化合物作为测试物质时,在整个pH范围内,管内和管间迁移时间变化(n = 6)的相对标准偏差(RSD)分别小于1%和2%。该涂层在十二烷基硫酸钠存在下相当稳定,这使得在低pH值下进行快速MEKC分离成为可能。当使用中性化合物作为测试物质时,在pH值为2至9的范围内,管内迁移时间变化(n = 6)的RSD小于2%。除了在低pH值下进行快速CZE和MEKC分离外,还可以通过CE-MS对碱性化合物进行分析。