Metabolite Measurement: Pitfalls to Avoid and Practices to Follow.
Annu Rev Biochem
; 86: 277-304, 2017 06 20.
Article
em En
| MEDLINE
| ID: mdl-28654323
ABSTRACT
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 acetonitrilemethanolwater) 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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MEDLINE
Assunto principal:
Espectrometria de Massas
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Espectroscopia de Ressonância Magnética
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Cromatografia Líquida
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Metabolômica
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Cromatografia Gasosa-Espectrometria de Massas
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article