Rambla José L, López-Gresa M P, Bellés J M, Granell Antonio
Instituto de Biología Molecular y Celular de Plantas, CSIC - Universidad Politécnica de Valencia, Ingeniero Fausto Elio s/n., 46022, Valencia, Spain.
Methods Mol Biol. 2015;1284:221-35. doi: 10.1007/978-1-4939-2444-8_11.
Metabolomics is a powerful discipline aimed at a comprehensive and global analysis of the metabolites present in a cell, tissue, or organism, and to which increasing attention has been paid in the last few years. Given the high diversity in physical and chemical properties of plant metabolites, not a single method is able to analyze them all.Here we describe two techniques for the profiling of two quite different groups of metabolites: polar and semi-polar secondary metabolites, including many of those involved in plant response to biotic and abiotic stress, and volatile compounds, which include those responsible of most of our perception of food flavor. According to these techniques, polar and semi-polar metabolites are extracted in methanol, separated by liquid chromatography (UPLC), and detected by a UV-VIS detector (PDA) and a time-of-flight (ToF) mass spectrometer. Volatile compounds, on the other hand, are extracted by headspace solid phase microextraction (HS-SPME), and separated and detected by gas chromatography coupled to mass spectrometry (GC-MS).
代谢组学是一门强大的学科，旨在对细胞、组织或生物体中存在的代谢物进行全面和整体的分析，在过去几年中受到了越来越多的关注。鉴于植物代谢物的物理和化学性质高度多样，没有一种单一的方法能够对它们全部进行分析。在这里，我们描述了两种用于分析两类截然不同代谢物的技术：极性和半极性次生代谢物，包括许多参与植物对生物和非生物胁迫反应的代谢物，以及挥发性化合物，其中包括构成我们对食物风味大部分感知的化合物。根据这些技术，极性和半极性代谢物在甲醇中提取，通过液相色谱（超高效液相色谱）分离，并由紫外可见检测器（光电二极管阵列检测器）和飞行时间质谱仪检测。另一方面，挥发性化合物通过顶空固相微萃取提取，并通过气相色谱-质谱联用（气相色谱-质谱）进行分离和检测。
