Liu Qian, Yang Yanmin, Yao Shouzhuo
State Key Lab of Chemo/Biosensing & Chemometrics, College of Chemistry & Chemical Engineering, Hunan University, Changsha 410082, China.
J Chromatogr A. 2008 Apr 11;1187(1-2):260-6. doi: 10.1016/j.chroma.2008.02.009. Epub 2008 Feb 8.
Semipermanent surfactant coatings are effective for the prevention of wall adsorption of proteins in CE. However, they often suffer from their unsatisfactory coating stability as they essentially degrade from the capillary walls after the surfactants are removed from the buffer. In this paper, we proposed a facile and universal method to improve the stability of semipermanent surfactant coatings based on addition of an oppositely charged surfactant into the coating. Didodecyldimethylammonium bromide (DDAB) and a gemini surfactant, 18-6-18, were used as the model semipermanent coatings, and sodium dodecyl sulfate (SDS) was chosen as their oppositely charged surfactant. SDS can strongly alter the packing parameter P of the cationic surfactants, and consequently mediates the coating stability. With the increase of SDS concentration in coating, the coating stability first dramatically increases due to the enlarged P, and then decreases due to the weakness of electrostatic interaction between the capillary wall and surfactant coating. At the proper SDS concentration, very stable coatings can be obtained that, even after rinsing under 138 kPa for 60 min, the reversed electroosmotic flow (EOF) only decreases by 3.6%. These SDS-enhanced coatings show excellent stability and reproducibility in protein separation (RSD of migration time <1.1% for run-to-run assay, n=9). Also, the high separation efficiency (>500,000 plates/m) and fine recovery of tested proteins indicate that these coatings are powerful in wall adsorption suppression. Finally, we found that the separation efficiency of protein was a more exact indicator for the coating stability than the traditional EOF magnitude.
半永久性表面活性剂涂层对于防止蛋白质在毛细管电泳中吸附到管壁上是有效的。然而,它们常常存在涂层稳定性不尽人意的问题,因为在表面活性剂从缓冲液中去除后,它们本质上会从毛细管壁上降解。在本文中,我们提出了一种简便通用的方法来提高基于在涂层中添加带相反电荷表面活性剂的半永久性表面活性剂涂层的稳定性。十二烷基二甲基溴化铵(DDAB)和一种双子表面活性剂18-6-18被用作典型的半永久性涂层,而十二烷基硫酸钠(SDS)被选为它们带相反电荷的表面活性剂。SDS能强烈改变阳离子表面活性剂的堆积参数P,从而调节涂层稳定性。随着涂层中SDS浓度的增加,由于P增大,涂层稳定性首先显著提高,然后由于毛细管壁与表面活性剂涂层之间静电相互作用减弱而降低。在适当的SDS浓度下,可以获得非常稳定的涂层,即使在138 kPa压力下冲洗60分钟后,反向电渗流(EOF)仅降低3.6%。这些SDS增强的涂层在蛋白质分离中表现出优异的稳定性和重现性(运行间测定迁移时间的相对标准偏差<1.1%,n = 9)。此外,高分离效率(>500,000塔板/米)和测试蛋白质的良好回收率表明这些涂层在抑制壁吸附方面很有效。最后,我们发现蛋白质的分离效率比传统的EOF大小更能准确地指示涂层稳定性。