Metabolite Measurement: Pitfalls to Avoid and Practices to Follow.

Metabolite Measurement: Pitfalls to Avoid and Practices to Follow.
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DOI:
10.1146/annurev-biochem-061516-044952
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发表时间:
2017-06-20
影响因子:
16.6
通讯作者:
Rabinowitz JD
Rabinowitz JD
中科院分区:
生物学1区
文献类型:
--
作者:
Lu W;Su X;Klein MS;Lewis IA;Fiehn O;Rabinowitz JD

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代谢物是参与能量转化和生物合成的小生物分子。由于代谢物的反应性、结构多样性和广泛的浓度范围,研究代谢本身就具有挑战性。在此,我们回顾了代谢组学中遇到的常见陷阱,并提供了获得准确代谢物测量的具体指南,重点是水溶性初级代谢物。我们展示了看似简单的样品制备方法如何引入系统误差(例如,由于代谢物之间的相互转化),以及如何正确选择淬火溶剂(例如,酸性乙腈:甲醇:水)可以减轻这些问题。我们讨论了每种常用分析平台的具体优势、缺陷和最佳实践:液相色谱-质谱(LC-MS)、气相色谱-质谱(GC-MS)、核磁共振(NMR)和酶分析。总之,这些信息提供了一个实用的知识库进行生物学信息代谢物测量。
Metabolites are the small biological molecules involved in energy conversion and biosynthesis. Studying metabolism is inherently challenging due to metabolites’ reactivity, structural diversity, and broad concentration range. Herein, we review the common pitfalls encountered in metabolomics and provide concrete guidelines for obtaining accurate metabolite measurements, focusing on water-soluble primary metabolites. We show how seemingly straightforward sample preparation methods can introduce systematic errors (e.g., owing to interconversion among metabolites) and how proper selection of quenching solvent (e.g., acidic acetonitrile:methanol:water) can mitigate such problems. We discuss the specific strengths, pitfalls, and best practices for each common analytical platform: liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), and enzyme assays. Together this information provides a pragmatic knowledge base for carrying out biologically informative metabolite measurements.
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