Negative mode sheathless capillary electrophoresis electrospray ionization-mass spectrometry for metabolite analysis of prokaryotes
- 作者：James L. Edwards ; Claire N. Chisolm ; Jonathan G. Shackman and Robert T. Kennedy
- 关键词：Metabolomics ; Capillary electrophoresis ; Electrospray ionisation ; Mass spectrometry
- 刊名：Journal of Chromatography A
- 出版年：2006
- 期刊代码：47_00219673
- 类别：ch
- 出版时间：17 February 2006
- 卷：1106
- 期：1-2
- 页码：80-88
- 文件大小：252 K
摘要
Capillary electrophoresis (CE) was coupled to negative mode electrospray ionisation-mass spectrometry (MS) for separation and detection of phosphorylated and acidic metabolites in extracts of prokaryotes. Unlike previous CE–MS systems for metabolite analysis, a sheathless interface was used to improve sensitivity. To accomplish this, the separation capillary was modified by creating a porous junction near the outlet where the electrospray voltage and cathodic voltage for CE were applied. The outlet of the capillary was pulled to a 5 μm inner diameter to form an electrospray emitter and had a frit fabricated near the exit to prevent clogging. During analysis pressure was applied at the inlet of the separation column to create sufficient flow towards the detector. Limits of detection for 19 metabolites in full scan mode ranged from 20 nM for ADP ribose to 2.5 μM for α-ketoglutarate for 40 nL injections. Extracts of Escherichia coli, strain DH5-α, were analyzed using this system. In full scan mode, 118 different metabolites were detected. Tandem mass spectrometry was also employed to attempt identification. Reproducible fragmentation of 19 parent peaks was found and 10 of these produced spectra that were consistent with identification obtained from matching to compounds in the MetaCyc database. These results demonstrate the utility of a sensitive CE–MS system for large scale metabolite detection in biological samples.