J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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Originally published In Press as doi:10.1194/jlr.D500026-JLR200 on November 21, 2005

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Journal of Lipid Research, Vol. 47, 431-439, February 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Methods

Measurement of stable isotopic enrichment and concentration of long-chain fatty acyl-carnitines in tissue by HPLC-MS

Dayong Sun, Melanie G. Cree, Xiao-jun Zhang, Elisabet Bøersheim and Robert R. Wolfe1

Metabolism Unit, Shriners Burn Hospital, University of Texas Medical Branch, Galveston, TX 77550

Published, JLR Papers in Press, November 21, 2005.

1 To whom correspondence should be addressed. e-mail: rwolfe{at}utmb.edu

We have developed a new method for the simultaneous measurements of stable isotopic tracer enrichments and concentrations of individual long-chain fatty acyl-carnitines in muscle tissue using ion-pairing high-performance liquid chromatography-electrospray ionization quadrupole mass spectrometry in the selected ion monitoring (SIM) mode. Long-chain fatty acyl-carnitines were extracted from frozen muscle tissue samples by acetonitrile/methanol. Baseline separation was achieved by reverse-phase HPLC in the presence of the volatile ion-pairing reagent heptafluorobutyric acid. The SIM capability of a single quadrupole mass analyzer allows further separation of the ions of interest from the sample matrixes, providing very clean total and selected ion chromatograms that can be used to calculate the stable isotopic tracer enrichment and concentration of long-chain fatty acyl-carnitines in a single analysis. The combination of these two separation techniques greatly simplifies the sample preparation procedure and increases the detection sensitivity. Applying this protocol to biological muscle samples proves it to be a very sensitive, accurate, and precise analytical tool.

Supplementary key words muscle tissue • ion-pairing high-performance liquid chromatography • high-performance liquid chromatography-mass spectrometry


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Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.