Extending metabolome coverage for untargeted metabolite profiling of adherent cultured hepatic cells

Analytical and Bioanalytical Chemistry
Juan C García-CañaverasAgustín Lahoz

Abstract

MS-based metabolite profiling of adherent mammalian cells comprises several challenging steps such as metabolism quenching, cell detachment, cell disruption, metabolome extraction, and metabolite measurement. In LC-MS, the final metabolome coverage is strongly determined by the separation technique and the MS conditions used. Human liver-derived cell line HepG2 was chosen as adherent mammalian cell model to evaluate the performance of several commonly used procedures in both sample processing and LC-MS analysis. In a first phase, metabolite extraction and sample analysis were optimized in a combined manner. To this end, the extraction abilities of five different solvents (or combinations) were assessed by comparing the number and the levels of the metabolites comprised in each extract. Three different chromatographic methods were selected for metabolites separation. A HILIC-based method which was set to specifically separate polar metabolites and two RP-based methods focused on lipidome and wide-ranging metabolite detection, respectively. With regard to metabolite measurement, a Q-ToF instrument operating in both ESI (+) and ESI (-) was used for unbiased extract analysis. Once metabolite extraction and analysis conditions were ...Continue Reading

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Citations

Jun 7, 2016·Scientific Reports·Juan Carlos García-CañaverasAgustín Lahoz
Oct 25, 2016·Current Opinion in Biotechnology·Jennifer Haggarty, Karl Ev Burgess
May 18, 2017·Electrophoresis·Juan Carlos García-CañaverasAgustín Lahoz
Jul 3, 2017·Cellular and Molecular Life Sciences : CMLS·Sarah HaytonRobert D Trengove
Aug 30, 2018·Archives of Toxicology·Matthias CuykxAdrian Covaci
Jun 20, 2020·Analytical and Bioanalytical Chemistry·Manhal MiliBénédicte Elena-Herrmann
Sep 16, 2020·Analytical Methods : Advancing Methods and Applications·Yun LuoHaijun Zhang
Sep 24, 2020·International Journal of Molecular Sciences·Aneta BalcerczykGilles J P Rautureau

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