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

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 t...

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Bibliographic Details
Published in:Journal of lipid research 2006-02, Vol.47 (2), p.431-439
Main Authors: Sun, Dayong, Cree, Melanie G, Zhang, Xiao-Jun, Bøersheim, Elisabet, Wolfe, Robert R
Format: Article
Language:English
Subjects:
Animals
Carbon Isotopes - analysis
Carnitine - analogs & derivatives
Carnitine - analysis
Carnitine - chemistry
Chromatography, High Pressure Liquid - methods
Deuterium - analysis
Fluorocarbons - chemistry
high-performance liquid chromatography-mass spectrometry
ion-pairing high-performance liquid chromatography
Male
Methods
muscle tissue
Muscle, Skeletal - chemistry
Palmitic Acid - blood
Palmitic Acid - chemistry
Palmitic Acid - metabolism
Palmitoylcarnitine - analysis
Palmitoylcarnitine - chemistry
Palmitoylcarnitine - standards
Rabbits
Reference Standards
Reproducibility of Results
Spectrometry, Mass, Electrospray Ionization - methods
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Summary: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.
ISSN:0022-2275
DOI:10.1194/jlr.D500026-JLR200