Carneiro Sónia, Pereira Rui, Rocha Isabel
IBB-Institute for Biotechnology and Bioengineering, Center of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal,
Methods Mol Biol. 2014;1152:197-207. doi: 10.1007/978-1-4939-0563-8_12.
Metabolome sample preparation is one of the key factors in metabolomics analyses. The quality of the metabolome data will depend on the suitability of the experimental procedures to the cellular system (e.g., yeast cells) and the analytical performance. Here, we summarize a protocol for metabolome analysis of yeast cells using gas chromatography-mass spectrometry (GC-MS). First, the main phases of a metabolomics analysis are identified: sample preparation, metabolite extraction, and analysis. We also provide an overview on different methods used to quench samples and extract intracellular metabolites from yeast cells. This protocol provides a detailed description of a GC-MS-based analysis of yeast metabolome, in particular for metabolites containing amino and/or carboxyl groups, which represent most of the compounds participating in the central carbon metabolism.
代谢组学样本制备是代谢组学分析的关键因素之一。代谢组学数据的质量将取决于实验程序对细胞系统（如酵母细胞）的适用性以及分析性能。在此，我们总结了一种使用气相色谱 - 质谱联用（GC-MS）对酵母细胞进行代谢组学分析的方案。首先，确定代谢组学分析的主要阶段：样本制备、代谢物提取和分析。我们还概述了用于淬灭样本以及从酵母细胞中提取细胞内代谢物的不同方法。本方案详细描述了基于GC-MS的酵母代谢组分析，特别是针对含有氨基和/或羧基的代谢物，这些代谢物代表了参与中心碳代谢的大多数化合物。
