Beckonert Olaf, Keun Hector C, Ebbels Timothy M D, Bundy Jacob, Holmes Elaine, Lindon John C, Nicholson Jeremy K
Department of Biomolecular Medicine, Faculty of Medicine, Imperial College London, Sir Alexander Fleming Building, South Kensington, London, UK.
Nat Protoc. 2007;2(11):2692-703. doi: 10.1038/nprot.2007.376.
Metabolic profiling, metabolomic and metabonomic studies mainly involve the multicomponent analysis of biological fluids, tissue and cell extracts using NMR spectroscopy and/or mass spectrometry (MS). We summarize the main NMR spectroscopic applications in modern metabolic research, and provide detailed protocols for biofluid (urine, serum/plasma) and tissue sample collection and preparation, including the extraction of polar and lipophilic metabolites from tissues. 1H NMR spectroscopic techniques such as standard 1D spectroscopy, relaxation-edited, diffusion-edited and 2D J-resolved pulse sequences are widely used at the analysis stage to monitor different groups of metabolites and are described here. They are often followed by more detailed statistical analysis or additional 2D NMR analysis for biomarker discovery. The standard acquisition time per sample is 4-5 min for a simple 1D spectrum, and both preparation and analysis can be automated to allow application to high-throughput screening for clinical diagnostic and toxicological studies, as well as molecular phenotyping and functional genomics.
代谢谱分析、代谢组学和代谢组学研究主要涉及使用核磁共振光谱和/或质谱（MS）对生物体液、组织和细胞提取物进行多组分分析。我们总结了现代代谢研究中核磁共振光谱的主要应用，并提供了生物体液（尿液、血清/血浆）和组织样本采集与制备的详细方案，包括从组织中提取极性和亲脂性代谢物。在分析阶段，诸如标准一维光谱、弛豫编辑、扩散编辑和二维J分辨脉冲序列等1H核磁共振光谱技术被广泛用于监测不同组别的代谢物，本文将对此进行描述。随后通常会进行更详细的统计分析或额外的二维核磁共振分析以发现生物标志物。对于简单的一维光谱，每个样本的标准采集时间为4 - 5分钟，样本制备和分析均可实现自动化，以便应用于临床诊断和毒理学研究的高通量筛选，以及分子表型分析和功能基因组学研究。
