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Methods Mol Biol. 2019;1859:171-184. doi: 10.1007/978-1-4939-8757-3_9.

Exploiting High-Resolution Mass Spectrometry for Targeted Metabolite Quantification and 13C-Labeling Metabolism Analysis.

Li Z1,2,3, Li Y4, Tang YJ5, Shui W6.

Author information

1
iHuman Institute, ShanghaiTech University, Shanghai, China.
2
Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
3
University of Chinese Academy of Sciences, Beijing, China.
4
College of Life Sciences, Nankai University, Tianjin, China.
5
Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, St. Louis, MO, USA.
6
iHuman Institute, ShanghaiTech University, Shanghai, China. shuiwq@shanghaitech.edu.cn.

Abstract

Quantification of targeted metabolites, especially trace metabolites and structural isomers, in complex biological materials is an ongoing challenge for metabolomics. In this chapter, we summarize high-resolution mass spectrometry-based approaches mainly used for targeted metabolite and metabolomics analysis, and then introduce an MS1/MS2-combined PRM workflow for quantification of central carbon metabolism intermediates, amino acids, and shikimate pathway-related metabolites. Major steps in the workflow, including cell culture, metabolite extraction, LC-MS analysis and data processing, are described. Furthermore, we adapt this new approach to a dynamic 13C-labeling experiment and demonstrate its unique advantage in capturing and correcting isotopomer labeling curves to facilitate nonstationary 13C-labeling metabolism analysis.

KEYWORDS:

13C-labeling metabolism analysis; High-resolution MS; Metabolite quantification; Parallel reaction monitoring (PRM)

PMID:
30421229
DOI:
10.1007/978-1-4939-8757-3_9
[Indexed for MEDLINE]

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