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Comparison of Five Extraction Methods for Intracellular Metabolites of Salmonella typhimurium

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Abstract

Salmonella enterica serovar typhimurium (S. typhimurium) causes food poisoning in human and animals. Its infection rate is the highest among all salmonella serotypes. Metabolomics is a potential way to study the pathogenesis of S. typhimurium via analysis of various small molecular substances. Due to the lack of a uniform protocol for the extraction of metabolites, we evaluated five commonly used extraction methods including cold methanol (CM), hot ethanol (HE), chloroform–methanol cocktail (CMC), perchloric acid (PCA), and alkali (AL) for their efficacy in extracting the intracellular metabolites of S. typhimurium. Samples were quenched in 60% methanol at − 40 °C, and then the five methods were used to extract the metabolites. After derivatization, all samples were analyzed on a gas chromatography–triple quadrupole mass spectrometry (GC–MS/MS). Our results suggest that CM and HE extraction methods provide the best compromise allowing identification of 98 and 95 metabolites in a single analysis. For targeted metabolome analysis, the optimal extraction method for alcohols and organic acids is HE. CMC preferentially extracted lipid metabolites. PCA is suitable for extraction of small molecular carbohydrates. The optimal extraction method for macromolecular carbohydrates is the CM method.

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Acknowledgements

This study is supported by the China Postdoctoral Science Foundation (Grant No. 2016M602691) and Sichuan Science & Technology Department Foundation (Grant No. 2017JY0240).

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  1. Sicheng Tian and Chuan Wang contribute equally to this study and are listed as first authors.

Authors and Affiliations

  1. Department of Laboratory Sciences of Public Health, West China School of Public Health and West China Fourth Hospital, Sichuan University, 16#, Section 3, South Renmin Road, West China, Chengdu, People’s Republic of China

    Sicheng Tian, Chuan Wang, Le Yang, Yunwen Zhang & Tian Tang

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  1. Sicheng Tian
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  2. Chuan Wang
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  3. Le Yang
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  4. Yunwen Zhang
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Correspondence to Tian Tang.

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Tian, S., Wang, C., Yang, L. et al. Comparison of Five Extraction Methods for Intracellular Metabolites of Salmonella typhimurium. Curr Microbiol 76, 1247–1255 (2019). https://doi.org/10.1007/s00284-019-01750-4

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  • DOI: https://doi.org/10.1007/s00284-019-01750-4

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