Lisec Jan, Schauer Nicolas, Kopka Joachim, Willmitzer Lothar, Fernie Alisdair R
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476 Potsdam-Golm, Germany.
Nat Protoc. 2006;1(1):387-96. doi: 10.1038/nprot.2006.59.
The concept of metabolite profiling has been around for decades, but technical innovations are now enabling it to be carried out on a large scale with respect to the number of both metabolites measured and experiments carried out. Here we provide a detailed protocol for gas chromatography mass spectrometry (GC-MS)-based metabolite profiling that offers a good balance of sensitivity and reliability, being considerably more sensitive than NMR and more robust than liquid chromatography-linked mass spectrometry. We summarize all steps from collecting plant material and sample handling to derivatization procedures, instrumentation settings and evaluating the resultant chromatograms. We also define the contribution of GC-MS-based metabolite profiling to the fields of diagnostics, gene annotation and systems biology. Using the protocol described here facilitates routine determination of the relative levels of 300-500 analytes of polar and nonpolar extracts in approximately 400 experimental samples per week per machine.
代谢物谱分析的概念已经存在了几十年，但技术创新使得它现在能够在大规模上进行，无论是在测量的代谢物数量还是进行的实验方面。在这里，我们提供了一个基于气相色谱 - 质谱联用（GC-MS）的代谢物谱分析的详细方案，该方案在灵敏度和可靠性之间取得了良好的平衡，比核磁共振（NMR）灵敏得多，比液相色谱 - 串联质谱更稳健。我们总结了从收集植物材料、样品处理到衍生化程序、仪器设置以及评估所得色谱图的所有步骤。我们还定义了基于GC-MS的代谢物谱分析在诊断、基因注释和系统生物学领域的贡献。使用这里描述的方案有助于每周每台机器在大约400个实验样品中常规测定300 - 500种极性和非极性提取物分析物的相对水平。
