Protocol abstract
Nature Protocols 2, - 2692 - 2703 (2007)
Published online: 25 October 2007 | doi:10.1038/nprot.2007.376
Subject Categories: Genomics and proteomics | Spectroscopy and structural analysis
Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts
Olaf Beckonert1, Hector C Keun1, Timothy M D Ebbels1, Jacob Bundy1, Elaine Holmes1, John C Lindon1 & Jeremy K Nicholson1
Abstract
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.
- Department of Biomolecular Medicine, Faculty of Medicine, Imperial College London, Sir Alexander Fleming Building, South Kensington, London, UK.
Correspondence to: Jeremy K Nicholson1 e-mail: j.nicholson@imperial.ac.uk
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A-Z product listing
- 1 ml(Genevac Ltd.)
- 1.5 ml(VWR International Ltd.)
- 2(Sigma-Aldrich)
- 3-d4 acid sodium salt(Sigma-Aldrich)
- 600 MHz Avance DRX NMR spectrometer(Bruker Biospin)
- 70% wt/wt(Sigma-Aldrich)
