Simultaneous determination of anionic intermediates for Bacillus subtilis metabolic pathways by capillary electrophoresis electrospray ionization mass spectrometry
Simultaneous determination of anionic intermediates for Bacillus subtilis metabolic pathways by capillary electrophoresis electrospray ionization mass spectrometry
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DOI：
10.1021/ac020064n
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发表时间：
2002-05-15
影响因子：
7.4
通讯作者：
Nishioka, T
中科院分区：
文献类型：
--
作者：
Soga, T;Ueno, Y;Nishioka, T
建立了毛细管电泳-电喷雾质谱联用技术同时测定阴离子代谢物的方法。为了防止电流下降的系统，电渗流（电渗流）逆转使用阳离子聚合物涂层毛细管是必不可少的。一个混合物含有32个标准品，包括羧酸，磷酸化羧酸，磷酸化磷酸，核苷酸，烟酰胺和黄素腺嘌呤辅酶的糖酵解和三羧酸循环途径的分离CE和选择性检测的四极质谱仪与鞘流电喷雾电离接口。分析的关键是使用阳离子聚合物涂覆的毛细管和由50 mM乙酸铵（pH 9.0）组成的电解质系统进行反相。方法的相对标准偏差为0.4%，迁移时间优于0.9%和5.4%之间的峰面积。这些代谢物的浓度检测限在0.3和6.7 mumol/L之间，压力注射为50 mbar，持续30 s（30 nL）;即，在信噪比为3的情况下，质量检测限范围为9至200 fmol。将该方法应用于枯草芽孢杆菌代谢中间产物的综合分析，可直接检测和定量27种阴离子代谢产物。
A method for simultaneous determination of anionic metabolites based on capillary electrophoresis (CE) coupled to electrospray ionization mass spectrometry is described. To prevent current drop by the system, electroosmotic flow (EOF) reversal by using a cationic polymer-coated capillary was indispensable. A mixture containing 32 standards including carboxylic acids, phosphorylated carboxylic acids, phosphorylated saccharides, nucleotides, and nicotinamide and flavin adenine coenzymes of glycolysis and the tricarboxylic acid cycle pathways were separated by CE and selectively detected by a quadrupole mass spectrometer with a sheath-flow electrospray ionization interface. Key to the analysis was EOF reversal using a cationic polymer-coated capillary and an electrolyte system consisting of 50 mM ammonium acetate, pH 9.0. The relative standard deviations of the method were better than 0.4% for migration times and between 0.9% and 5.4% for peak areas. The concentration detection limits for these metabolites were between 0.3 and 6.7 mumol/L with pressure injection of 50 mbar for 30 s (30 nL); i.e., mass detection limits ranged from 9 to 200 fmol, at a signal-to-noise ratio of 3. This method was applied to the comprehensive analysis of metabolic intermediates extracted from Bacillus subtilis, and 27 anionic metabolites could be directly detected and quantified.