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Metabolites 2014, 4(2), 142-165;

Application of Stable Isotope-Assisted Metabolomics for Cell Metabolism Studies

2,†,* and 1,*
Department of Energy, Environmental and Chemical Engineering, Washington University, St. Louis, MO 63130, USA
Plant Metabolomics Group, Institute of Plant Physiology and Ecology, Shanghai Institute for Biological Sciences, CAS, Shanghai 20032, China
Current address: Analytical Services, Lipospectrum LLC, 3830 Washington Blvd, Suite 121-122, St. Louis, MO 63108, USA.
Authors to whom correspondence should be addressed.
Received: 13 January 2014 / Revised: 18 March 2014 / Accepted: 20 March 2014 / Published: 31 March 2014
(This article belongs to the Special Issue Cell and Tissue Metabolomics)
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The applications of stable isotopes in metabolomics have facilitated the study of cell metabolisms. Stable isotope-assisted metabolomics requires: (1) properly designed tracer experiments; (2) stringent sampling and quenching protocols to minimize isotopic alternations; (3) efficient metabolite separations; (4) high resolution mass spectrometry to resolve overlapping peaks and background noises; and (5) data analysis methods and databases to decipher isotopic clusters over a broad m/z range (mass-to-charge ratio). This paper overviews mass spectrometry based techniques for precise determination of metabolites and their isotopologues. It also discusses applications of isotopic approaches to track substrate utilization, identify unknown metabolites and their chemical formulas, measure metabolite concentrations, determine putative metabolic pathways, and investigate microbial community populations and their carbon assimilation patterns. In addition, 13C-metabolite fingerprinting and metabolic models can be integrated to quantify carbon fluxes (enzyme reaction rates). The fluxome, in combination with other “omics” analyses, may give systems-level insights into regulatory mechanisms underlying gene functions. More importantly, 13C-tracer experiments significantly improve the potential of low-resolution gas chromatography-mass spectrometry (GC-MS) for broad-scope metabolism studies. We foresee the isotope-assisted metabolomics to be an indispensable tool in industrial biotechnology, environmental microbiology, and medical research. View Full-Text
Keywords: 13C-fingerprinting; flux; GC-MS; isotopologue; mass-to-charge; regulatory mechanisms 13C-fingerprinting; flux; GC-MS; isotopologue; mass-to-charge; regulatory mechanisms

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You, L.; Zhang, B.; Tang, Y.J. Application of Stable Isotope-Assisted Metabolomics for Cell Metabolism Studies. Metabolites 2014, 4, 142-165.

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