Global urinary metabolic profiling procedures using gas chromatography

Global urinary metabolic profiling procedures using gas chromatography–mass spectrometry
Eric Chun Yong Chan,1 Kishore Kumar Pasikanti1, 2 & Jeremy K Nicholson3

Journal name: Nature Protocols 
Volume: 6, Pages: 1483–1499 
Year published: (2011) DOI: doi:10.1038/nprot.2011.375 
Published online 08 September 2011

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.

原文：http://www.nature.com/nprot/journal/v6/n10/abs/nprot.2011.375.html