1. Nontargeted, Discovery-Based Profiling of the Yeast Metabolome Using QTOF LC-MS and LC-MS-MS.
- Author
-
Jenkins, Stefan, Fischer, Steven M., and Sana, Theodore R.
- Subjects
- *
LIQUID chromatography-mass spectrometry , *YEAST , *IMMUNOSUPPRESSIVE agents , *CALCIUM , *TIME-of-flight mass spectrometry , *CALCINEURIN - Abstract
This article describes a nontargeted, discovery-based approach for analyzing the yeast metabolome in response to calcium and immunosuppressant drug treatment, using multiple liquid chromatography (LC) conditions and quadrupole time-of-flight mass spectrometry (QTOFMS) analysis. Yeast cultures were exposed to calcium only or calcium and one of two immunosuppressant drugs with the goal of perturbing the calcineurin- and any other calcium- or immunosuppressant-responsive pathways. Compounds were extracted from the raw data using an unbiased extraction algorithm. These results were subsequently analyzed using multidimensional statistical analysis and data visualization software and reduced to a list of compounds that were differentially expressed according to culture condition. Accurate-mass-based identifications were done using an endogenous metabolite database. MS-MS spectra were acquired for a subset of compounds, allowing for matching at multiple fragmentation energies against an endogenous metabolite library to provide higher confidence identifications. [ABSTRACT FROM AUTHOR]
- Published
- 2013