AJP - Endocrinology and Metabolism, Vol 263, Issue 3 E584-E596, Copyright © 1992 by American Physiological Society

ARTICLES

Stable isotope tracer analysis by GC-MS, including quantification of isotopomer effects

J. Rosenblatt, D. Chinkes, M. Wolfe and R. R. Wolfe
Metabolism Unit, Shriners Burns Institute and Office Biomathematics, Galveston, Texas.

In metabolic tracer studies it is frequently useful to infuse tracers that are differently labeled variants of the same molecule. These tracers are known as isotopomers. Analysis of the enrichment of each isotopic analogue can be accomplished by gas chromatography-mass spectrometry (GC-MS). However, the raw GC-MS data must be corrected to give the information required. This paper addresses how to transform the raw GC-MS data, consisting of relative abundance ratios at specific ion masses, into relative molar ratios of tracer and tracee molecules. Several correction factors are necessary. First, the background must be measured and corrected for, since it is always present in the sample. Second, the abundances in the spectrum of the labeled molecule are different from those in the unlabeled molecule, and this proportionality "skew" is corrected. A third correction factor accounts for the overlapping spectra of two or more isotopomers that cannot be measured independently. The final correction removes the "double vision" effect that may appear in some spectra due to the presence of (M - H)+ species.

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