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Stable Isotope-Labeled Tracers for Metabolic Pathway Elucidation by GC-MS and FT-MS

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Mass Spectrometry in Metabolomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1198))

Abstract

Advances in analytical methodologies, principally nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS), over the last decade have made large-scale analysis of the human metabolome a reality. This is leading to the reawakening of the importance of metabolism in human diseases, particularly widespread metabolic diseases such as cancer, diabetes, and obesity. Emerging NMR and MS atom-tracking technologies and informatics are poised to revolutionize metabolomics-based research because they deliver the high information throughput (HIT) that is needed for deciphering systems biochemistry. In particular, stable isotope-resolved metabolomics (SIRM) enables unambiguous tracking of individual atoms through compartmentalized metabolic networks in a wide range of experimental systems, including human subjects. MS offers a wide range of instrumental capabilities involving different levels of initial capital outlay and operating costs, ranging from gas-chromatography (GC) MS that is affordable by many individual laboratories to the HIT-supporting Fourier-transform (FT) class of MS that rivals NMR in cost and infrastructure support. This chapter focuses on sample preparation, instrument, and data processing procedures for these two extremes of MS instrumentation used in SIRM.

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Acknowledgements

This work was supported in part by National Science Foundation EPSCoR grants# EPS-0447479 (T.W.M.F.) and EPS-0132295 (R.J. Wittebort), NIH NCRR 5P20RR018733 (D.M. Miller, A.N.L.), 1R01CA118434-01A2, 1RO1CA101199-01, R01ES-022191-01 and 3R01CA118434-02S1 (T.W.M.F.), R21CA133-668-02, and P01CA163223-01A1 (A.N.L.) from the National Cancer Institute, the University of Louisville CTSPGP/ARRA grants 20044, the Kentucky Lung Cancer Research Program (OGMB090354B1 and OGMB101380) (T.W.M.F. and A.N.L.), the Robert W. Rounsavall Jr. Family Foundation, and the Kentucky Challenge for Excellence. We thank Jin Lian Tan, Alex Belshoff, Katherine Sellers, and Radhika Burra for technical assistance and/or comments on the manuscript.

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Authors and Affiliations

  1. Graduate Center of Toxicology, University of Kentucky, Biopharm Complex, 789 S. Limestone St., Lexington, KY, 40536, USA

    Richard M. Higashi, Teresa W.-M. Fan & Andrew N. Lane

  2. Diabetes and Obesity Center, University of Louisville, 580 S. Preston St., Louisville, KY, 40202, USA

    Pawel K. Lorkiewicz

  3. Molecular and Cellular Biochemistry, University of Kentucky, CC434 Roach Building, 800 Rose Street, Lexington, KY, 40536-0093, USA

    Hunter N. B. Moseley

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  1. Richard M. Higashi
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  2. Teresa W.-M. Fan
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  3. Pawel K. Lorkiewicz
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  4. Hunter N. B. Moseley
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  5. Andrew N. Lane
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Correspondence to Richard M. Higashi .

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  1. University of Washington Fred Hutchinson Cancer Rsrch Ctr, Seattle, Washington, USA

    Daniel Raftery

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Higashi, R.M., Fan, T.WM., Lorkiewicz, P.K., Moseley, H.N.B., Lane, A.N. (2014). Stable Isotope-Labeled Tracers for Metabolic Pathway Elucidation by GC-MS and FT-MS. In: Raftery, D. (eds) Mass Spectrometry in Metabolomics. Methods in Molecular Biology, vol 1198. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1258-2_11

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  • DOI: https://doi.org/10.1007/978-1-4939-1258-2_11

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