Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts
Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts
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DOI：
10.1038/nprot.2007.376
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Nicholson, Jeremy K.
中科院分区：
文献类型：
--
作者：
Beckonert, Olaf;Keun, Hector C.;Nicholson, Jeremy K.
代谢谱、代谢组学和代谢组学研究主要涉及使用NMR光谱和/或质谱（MS）对生物液体、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的主要NMR光谱应用，并提供了生物流体（尿液，血清/血浆）和组织样品收集和制备的详细协议，包括从组织中提取极性和亲脂性代谢物。1H-1 NMR光谱技术，如标准1D光谱、弛豫编辑、扩散编辑和2D J分辨脉冲序列，广泛用于分析阶段，以监测不同的代谢物组，并在此进行描述。它们通常随后进行更详细的统计分析或额外的2D NMR分析以发现生物标志物。对于简单的一维光谱，每个样品的标准采集时间为4-5分钟，并且制备和分析都可以自动化，以允许应用于临床诊断和毒理学研究的高通量筛选，以及分子表型和功能基因组学。
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. H-1 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.