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无鞘毛细管电泳-质谱联用技术用于生物样品中带电代谢物的分析

Sheathless Capillary Electrophoresis-Mass Spectrometry for the Profiling of Charged Metabolites in Biological Samples.

作者信息

Ramautar Rawi

机构信息

Leiden Academic Center for Drug Research, Leiden University, Leiden, The Netherlands.

出版信息

Methods Mol Biol. 2018;1738:183-192. doi: 10.1007/978-1-4939-7643-0_12.

Abstract

Capillary electrophoresis (CE) is well suited for the profiling of highly polar and charged metabolites as compounds are separated on the basis of their charge-to-size ratio. The protocol presented here is based on using a recently developed sheathless interfacing design, i.e., a porous tip interface, for coupling CE to electrospray ionization mass spectrometry (MS). It is demonstrated that sheathless CE-MS employing a bare fused-silica capillary at low-pH separation conditions can be used for the profiling of both cationic and anionic metabolites by only switching the MS detection and electrophoretic separation voltage polarity. The proposed sheathless CE-MS protocol allows efficient and sensitive profiles to be obtained for a broad array of charged metabolites, including amino acids, organic acids, nucleotides, and sugar phosphates, in various biological samples, such as urine and extracts of the glioblastoma cell line.

摘要

毛细管电泳(CE)非常适合用于分析高极性和带电代谢物,因为化合物是根据其电荷与大小之比进行分离的。本文介绍的方案基于使用一种最近开发的无鞘接口设计,即多孔尖端接口,用于将CE与电喷雾电离质谱(MS)联用。结果表明,在低pH分离条件下使用裸露熔融石英毛细管的无鞘CE-MS,仅通过切换MS检测和电泳分离电压极性,就可用于分析阳离子和阴离子代谢物。所提出的无鞘CE-MS方案能够在各种生物样品(如尿液和成胶质细胞瘤细胞系提取物)中,高效且灵敏地分析包括氨基酸、有机酸、核苷酸和糖磷酸盐在内的多种带电代谢物。

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