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采用发光二极管诱导荧光检测的多毛细管凝胶电泳分析复合碳水化合物的样品制备

Sample preparation for the analysis of complex carbohydrates by multicapillary gel electrophoresis with light-emitting diode induced fluorescence detection.

作者信息

Olajos Marcell, Hajós Péter, Bonn Guenther K, Guttman András

机构信息

Horvath Laboratory of Bioseparation Sciences, Institute of Analytical Chemistry and Radiochemistry, University of Innsbruck, Innsbruck, Austria.

出版信息

Anal Chem. 2008 Jun 1;80(11):4241-6. doi: 10.1021/ac8002598. Epub 2008 May 7.

Abstract

This paper evaluates various sample preparation methods for multicapillary gel electrophoresis based glycan analysis to support electrokinetic injection. First the removal of excess derivatization reagent is discussed. Although the Sephadex G10 filled multiscreen 96-well filter plate and Sephadex G10 filled pipet tips enabled increased analysis sensitivity, polyamide DPA-6S pipet tips worked particularly well. In this latter case an automated liquid handling system was used to increase purification throughput, necessary to feed the multicapillary electrophoresis unit. Problems associated with the high glucose content of such biological samples as normal human plasma were solved by applying ultrafiltration. Finally, a volatile buffer system was developed for exoglycosidase-based carbohydrate analysis.

摘要

本文评估了基于多毛细管凝胶电泳的聚糖分析中用于支持电动进样的各种样品制备方法。首先讨论了去除过量衍生试剂的方法。尽管填充有Sephadex G10的多筛96孔滤板和填充有Sephadex G10的移液器吸头能够提高分析灵敏度,但聚酰胺DPA-6S移液器吸头的效果特别好。在后一种情况下,使用了自动液体处理系统来提高纯化通量,这对于向多毛细管电泳单元供样是必要的。通过应用超滤解决了诸如正常人血浆等生物样品中高葡萄糖含量相关的问题。最后,开发了一种用于基于外切糖苷酶的碳水化合物分析的挥发性缓冲系统。

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