Zhang Wei, Hankemeier Thomas, Ramautar Rawi
Biomedical Microscale Analytics, Leiden Academic Center for Drug Research, Leiden University, Leiden, The Netherlands.
Methods Mol Biol. 2019;1972:165-172. doi: 10.1007/978-1-4939-9213-3_11.
Capillary electrophoresis-mass spectrometry (CE-MS) is a strong separation technique for the highly efficient and selective analysis of polar and charged metabolites in biological samples. The CE approach is especially suited for the analysis of limited sample amounts due to its nanoliter injections from only a few microliters of material in the sample injection vial. In this protocol, a CE-MS strategy is outlined for the profiling of cationic metabolites in biomass-limited samples using a small amount of human hepatocellular carcinoma (HepG2) cells as a model system. By employing a sheathless interfacing design for coupling CE to MS, it is shown that information-rich profiles for cationic metabolites can be obtained when working with a starting amount of 10,000 HepG2 cells and even lower. Overall, the proposed CE-MS-based analytical workflow may be considered a useful tool for biomass-limited metabolomics studies.
毛细管电泳-质谱联用(CE-MS)是一种强大的分离技术,用于高效、选择性地分析生物样品中的极性和带电代谢物。由于在样品进样瓶中仅需从几微升材料中进行纳升进样,CE方法特别适合于分析少量样品。在本方案中,概述了一种CE-MS策略,以少量人肝癌(HepG2)细胞作为模型系统,对生物量有限的样品中的阳离子代谢物进行分析。通过采用用于CE与MS联用的无鞘接口设计,结果表明,当起始细胞量为10000个HepG2细胞甚至更低时,仍可获得富含信息的阳离子代谢物谱。总体而言,所提出的基于CE-MS的分析工作流程可被视为生物量有限的代谢组学研究的有用工具。