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. 2020 Aug 19;7:211. doi: 10.3389/fmolb.2020.00211

FIGURE 1.

FIGURE 1

A comprehensive optimization of sample preparation for GC-MS/MS-based metabolomics in U. maydis. Cell cultivations were performed in System Duetz® 24-deep well plates at 30°C in minimal media. To instantly stop cellular metabolism, both fast filtration and quick quenching methods were tested. Then, cells were washed one (x1), two (x2), and three (x3) times with cold saline buffer to remove the extracellular components from culture medium. Eight different strategies combining chemical and mechanical disruption methods were tested for extraction efficiency. Finally, metabolites were derivatized at 37°C, 60°C, or 80°C. All experiments were performed with three biological replicates. Abbreviations: SB, Saline Buffer; CE, cold EtOH; CM, cold MeOH; CC, cold chloroform/MeOH/water mixture; HE, hot EtOH; HM, hot MeOH; SE, EtOH with sonication; SM, MeOH with sonication; SC, chloroform/MeOH/water mixture with sonication.