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A more recent version of this article appeared on February 1, 2006

Papers In Press, published online ahead of print November 21, 2005
J. Lipid Res., doi:10.1194/jlr.D500026-JLR200
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Submitted on August 3, 2005
Revised on October 31, 2005
Accepted on November 21, 2005

Measurement of stable Isotopic enrichment and concentration of long chain fatty acyl-carnitines in tissue by Ion-pairing HPLC / MS

Dayong Sun, Melanie G. Cree, Xiao-jun Zhang, Elisabet Boersheim, and Robert R. Wolfe

Metabolism Unit, University of Texas Medical Branch, Galveston, TX 77550

Corresponding Author: daysun{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. Base-line separation was achieved by a reversed phase HPLC in the presence of the volatile ion-pairing reagent, heptafluorobutyric acid (HFBA). 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.


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