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GC-MS profiling of leukemia cells: an optimized preparation protocol for the intracellular metabolome

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Abstract

Sample preparation is an important step in the untargeted cell metabolomics workflow. In this study, a gas chromatography-mass spectrometry (GC-MS)-based metabolome preparation protocol was optimized with the aim of achieving global, reliable and reproducible intracellular metabolite profiling of leukemia cells. We have evaluated and compared 8 sample preparation methods: 4 extraction solvents (i.e., pure methanol, aqueous acetonitrile, methanol/methanol/water and aqueous methanol) coupled with 2 common trimethylsilylation reagents (i.e., N,O-bis(trimethylsilyl)trifluoroacetamide and N-methyl-N-(trimethylsilyl)trifluoroacetamide). The extraction solvents and derivatization agents were evaluated for their suitability for leukemia cells using criteria based on the number of total detected metabolites, extraction efficiency and repeatability. Based on the overall performance, methanol/methanol/water was chosen as the optimal extraction solvent for covering the highest number of intracellular metabolites with the best reproducibility. The derivatization agent N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) proved to be more suitable for profiling of the leukemia intracellular metabolome with better repeatability than N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA). This optimized method provides a robust, efficient and reproducible way to gather information on the metabolome of a leukemia cell. It also offers a solid basis to further understand the physiology of leukemia cells and unravel the mechanism of leukemia’s multidrug resistance from the perspective of metabolomics.

Graphical abstract: GC-MS profiling of leukemia cells: an optimized preparation protocol for the intracellular metabolome

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Publication details

The article was received on 03 Nov 2017, accepted on 01 Feb 2018 and first published on 01 Feb 2018


Article type: Paper
DOI: 10.1039/C7AY02578E
Citation: Anal. Methods, 2018, Advance Article
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    GC-MS profiling of leukemia cells: an optimized preparation protocol for the intracellular metabolome

    Y. He, Z. M. Zhang, P. Ma, H. C. Ji and H. M. Lu, Anal. Methods, 2018, Advance Article , DOI: 10.1039/C7AY02578E

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