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Translational Bioinformatics for Therapeutic Development pp 301–313Cite as

Sample Preparation and Data Analysis for NMR-Based Metabolomics

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2194))

Abstract

NMR spectroscopy has become one of the preferred analytical techniques for metabolomics studies due to its inherent nondestructive nature, ability to identify and quantify metabolites simultaneously in a complex mixture, minimal sample preparation requirement, and high degree of experimental reproducibility. NMR-based metabolomics studies involve the measurement and multivariate statistical analysis of metabolites present in biological samples such as biofluids, stool/feces, intestinal content, tissue, and cell extracts by high-resolution NMR spectroscopy—the goal then is to identify and quantify metabolites and evaluate changes of metabolite concentrations in response to some perturbation. Here we describe methodologies for NMR sample preparation of biofluids (serum, saliva, and urine) and stool/feces, intestinal content, and tissues for NMR experiments including extraction of polar metabolites and application of NMR in metabolomics studies. One dimensional (1D) 1H NMR experiments with different variations such as pre-saturation, relaxation-edited, and diffusion-edited are routinely acquired for profiling and metabolite identification and quantification. 2D homonuclear 1H-1H TOCSY and COSY, 2D J-resolved, and heteronuclear 1H-13C HSQC and HMBC are also performed to assist with metabolite identification and quantification. The NMR data are then subjected to targeted and/or untargeted multivariate statistical analysis for biomarker discovery, clinical diagnosis, toxicological studies, molecular phenotyping, and functional genomics.

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Authors and Affiliations

  1. Department of Chemistry, Pennsylvania State University, University Park, PA, USA

    Tapas K. Mal

  2. Department of Veterinary and Biomedical Sciences, Pennsylvania State University, University Park, PA, USA

    Yuan Tian & Andrew D. Patterson

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  1. Tapas K. Mal
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  2. Yuan Tian
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  3. Andrew D. Patterson
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Correspondence to Tapas K. Mal .

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Editors and Affiliations

  1. Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA

    Joseph Markowitz

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Mal, T.K., Tian, Y., Patterson, A.D. (2021). Sample Preparation and Data Analysis for NMR-Based Metabolomics. In: Markowitz, J. (eds) Translational Bioinformatics for Therapeutic Development. Methods in Molecular Biology, vol 2194. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0849-4_16

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  • DOI: https://doi.org/10.1007/978-1-0716-0849-4_16

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