Extraction, LC/MS analysis and stress comparison of the Saccharomyces cerevisiae metabolome

We describe a non-targeted discovery-based metabolomics study of S.cerevisiae\ud using a novel metabolite extraction protocol, an LC-QTOF MS, and a variety of data\ud analysis techniques. A thorough, multi-step optimization of the extraction protocol was\ud completed prior to carrying out stress comparison experiments. With the efficacy of the\ud novel extraction protocol established, stress comparison experiments were carried out\ud to study the yeast response to calcium and immunosuppressant drug stress, the latter\ud effected with exposure to either FK506 or Cyclosporin A. Raw data was filtered,\ud baselined and deconvoluted. All entities found in the initial raw data were then aligned\ud and statistically analyzed in order to determine entities that showed differential\ud abundance across data sets. Database tools and MS/MS analysis were also used in\ud order to identify compounds and begin the transition from a non-targeted to a targeted\ud metabolomics workflow