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

  • Richard M. Higashi
  • Teresa W.-M. Fan
  • Pawel K. Lorkiewicz
  • Hunter N. B. Moseley
  • Andrew N. Lane
Part of the Methods in Molecular Biology book series (MIMB, volume 1198)


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.

Key words

Metabolomics Stable isotope Mass spectrometry FT-MS GC-MS 



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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Richard M. Higashi
    • 1
  • Teresa W.-M. Fan
    • 1
  • Pawel K. Lorkiewicz
    • 2
  • Hunter N. B. Moseley
    • 3
  • Andrew N. Lane
    • 1
  1. 1.Graduate Center of ToxicologyUniversity of KentuckyLexingtonUSA
  2. 2.Diabetes and Obesity CenterUniversity of LouisvilleLouisvilleUSA
  3. 3.Molecular and Cellular BiochemistryUniversity of KentuckyLexingtonUSA

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