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Optimization of extraction and analytical protocol for mass spectrometry‐based metabolomics analysis of hepatotoxicity
Author(s) -
Yang Rui,
Zhao Qi,
Hu DanDan,
Xiao XueRong,
Li Fei
Publication year - 2018
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4359
Subject(s) - chemistry , chromatography , metabolomics , extraction (chemistry) , mass spectrometry , high performance liquid chromatography , acetonitrile , liquid chromatography–mass spectrometry , quantitative analysis (chemistry) , liver injury , reproducibility , pharmacology , medicine
Drug‐induced liver injury is a clinically leading side‐effect of drugs. In the present study, a liquid chromatography mass spectrometry‐based metabolomics protocol was optimized for extraction and analysis of endogenous metabolites from liver tissue during hepatotoxicity. Various extraction solutions, resuspension solutions, extraction folds and dissolution methods for the supernatant were compared using the number of extracted total ions, relative response and relative extraction efficiency of targeted metabolites from liver tissue. The polar and nonpolar endogenous metabolites associated with liver injury were analyzed by hydrophilic interaction chromatography and reversed‐phase liquid chromatography with UPLC–QTOFMS. The results indicated that extraction with 10‐fold 50% acetonitrile in water and the supernatant diluted (1:1) with 100% acetonitrile rather than resuspension was the optimal extraction protocol. Subsequently, the optimized method was able to examine the change in metabolites in mouse liver tissue resulting from treatment with a toxic natural product, toosendanin. Taken together, the optimized extraction and analytical protocol provides high reliability and reproducibility for polar and nonpolar metabolites in liver tissue and may be suitable for metabolomics analysis of liver injury induced by drugs or chemicals.