Protocol | Published:

Global urinary metabolic profiling procedures using gas chromatography–mass spectrometry

Nature Protocols volume 6, pages 14831499 (2011) | Download Citation

Abstract

The role of urinary metabolic profiling in systems biology research is expanding. This is because of the use of this technology for clinical diagnostic and mechanistic studies and for the development of new personalized health care and molecular epidemiology (population) studies. The methodologies commonly used for metabolic profiling are NMR spectroscopy, liquid chromatography mass spectrometry (LC/MS) and gas chromatography–mass spectrometry (GC/MS). In this protocol, we describe urine collection and storage, GC/MS and data preprocessing methods, chemometric data analysis and urinary marker metabolite identification. Results obtained using GC/MS are complementary to NMR and LC/MS. Sample preparation for GC/MS analysis involves the depletion of urea via treatment with urease, protein precipitation with methanol, and trimethylsilyl derivatization. The protocol described here facilitates the metabolic profiling of 400–600 metabolites in 120 urine samples per week.

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Acknowledgements

GC/TOFMS method development was supported by the National University of Singapore (NUS) grant R-148-000-100-112 and National Medical Research Council grant R-176-000-119-213 provided to E.C.Y.C. GC/TOFMS was sponsored by the NUS grant R-279-000-249-646. P.K.K. is supported by a NUS President's Graduate fellowship.

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Affiliations

  1. Department of Pharmacy, Faculty of Science, National University of Singapore, Singapore.

    • Eric Chun Yong Chan
    •  & Kishore Kumar Pasikanti
  2. GlaxoSmithKline R&D China, Singapore Research Center, Biopolis at One-North, Singapore.

    • Kishore Kumar Pasikanti
  3. Biomolecular Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, UK.

    • Jeremy K Nicholson

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Contributions

E.C.Y.C. and P.K.K. designed the protocol and conducted the initial study; E.C.Y.C., P.K.K. and J.K.N. wrote the manuscript; J.K.N. gave conceptual advice.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Eric Chun Yong Chan.

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DOI

https://doi.org/10.1038/nprot.2011.375

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