Biomedical Microscale Analytics, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands.
Methods Mol Biol. 2023;2571:105-114. doi: 10.1007/978-1-0716-2699-3_10.
The simultaneous analysis of cationic and anionic metabolites using capillary electrophoresis-mass spectrometry (CE-MS) has been considered challenging, as often two different analytical methods are required. Although CE-MS methods for cationic metabolite profiling have already shown good performance metrics, the profiling of anionic metabolites often results in relatively low sensitivity and poor repeatability caused by problems related to unstable electrospray and corona discharge when using reversed CE polarity and detection by MS in negative ionization mode. In this protocol, we describe a chemical derivatization procedure that provides a permanent positive charge to acidic metabolites, thereby allowing us to profile anionic metabolites by CE-MS using exactly the same separation conditions as employed for the analysis of basic metabolites. The utility of the overall approach is demonstrated for the analysis of energy metabolism-related metabolites in low numbers of HepG2 cells.
使用毛细管电泳-质谱联用（CE-MS）同时分析阳离子和阴离子代谢物一直具有挑战性，因为通常需要两种不同的分析方法。尽管已经有用于阳离子代谢物分析的 CE-MS 方法，但在使用反相 CE 极性和负电离模式下的 MS 检测时，由于电喷雾和电晕放电不稳定相关问题，阴离子代谢物的分析往往导致相对较低的灵敏度和较差的重现性。在本方案中，我们描述了一种化学衍生化程序，该程序可为酸性代谢物提供永久正电荷，从而使我们能够通过 CE-MS 对阴离子代谢物进行分析，而使用的分离条件与碱性代谢物的分析完全相同。该方法在分析低数量 HepG2 细胞中的能量代谢相关代谢物方面的应用得到了证明。
