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Improving repeatability of capillary electrophoresis—a critical comparison of ten different capillary inner surfaces and three criteria of peak identification

机译:提高毛细管电泳的可重复性—十种不同毛细管内表面的严格比较和峰鉴定的三个标准

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摘要

A poor repeatability of migration times caused by the fluctuations of electroosmotic flow (EOF) is an inherent weakness of capillary electrophoresis. Most researchers endeavor to prevent this problem using relative migration times or various capillary coatings which are expensive and not easy in comparison. Herein, we present an original approach to this problem: we apply a model sample designed to induce significant EOF instability, in order to critically compare ten capillary types with different physicochemical characteristics. Moreover, we accompany capillary modification with the evaluation of various criteria of peak identification: migration time, migration times ratio, and electrophoretic mobility. Our results show a great effectiveness of a dynamic coating in the stabilization of migration times, with the average RSD(%) value reduced from 3.5% (bare silica capillary) down to 0.5%. The good outcomes were also obtained for the surfactant-modified silica and amine capillaries. For the capillaries exhibiting significant instability of EOF, electrophoretic mobility turned out to be a more universal and reliable criterion of peak identification than relative migration time. It can be explained by an intrinsic dependency of migration times ratio on EOF change, which should always be considered during the selection of an internal standard.Electronic supplementary materialThe online version of this article (doi:10.1007/s00216-017-0382-y) contains supplementary material, which is available to authorized users.
机译:由电渗流量(EOF)的波动引起的迁移时间重复性差是毛细管电泳的固有弱点。大多数研究人员都试图使用相对迁移时间或各种昂贵的毛细管涂层来防止此问题,并且比较起来并不容易。本文中,我们提出了解决此问题的原始方法:我们应用了设计为引起明显的EOF不稳定性的模型样本,以严格比较具有不同理化特性的十种毛细管类型。此外,我们在毛细管修饰的同时对各种峰鉴定标准进行了评估:迁移时间,迁移时间比率和电泳迁移率。我们的结果显示出动态涂层在稳定迁移时间方面的巨大功效,平均RSD(%)值从3.5%(裸二氧化硅毛细管)降至0.5%。表面活性剂改性的二氧化硅和胺毛细管也获得了良好的结果。对于表现出明显的EOF不稳定的毛细管,电泳迁移率是比相对迁移时间更通用和可靠的峰识别标准。可以用迁移时间比率对EOF变化的内在依赖性来解释,这在选择内标时应始终予以考虑。电子补充材料本文的在线版本(doi:10.1007 / s00216-017-0382-y)包含补充材料,授权用户可以使用。

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