出版社：American Society for Biochemistry and Molecular Biology
摘要：We have developed a new method for the simultaneous measurementsof stable isotopic tracer enrichments and concentrations ofindividual long-chain fatty acyl-carnitines in muscle tissueusing ion-pairing high-performance liquid chromatography-electrosprayionization quadrupole mass spectrometry in the selected ionmonitoring (SIM) mode. Long-chain fatty acyl-carnitines wereextracted from frozen muscle tissue samples by acetonitrile/methanol.Baseline separation was achieved by reverse-phase HPLC in thepresence of the volatile ion-pairing reagent heptafluorobutyricacid. The SIM capability of a single quadrupole mass analyzerallows further separation of the ions of interest from the samplematrixes, providing very clean total and selected ion chromatogramsthat can be used to calculate the stable isotopic tracer enrichmentand concentration of long-chain fatty acyl-carnitines in a singleanalysis. The combination of these two separation techniquesgreatly simplifies the sample preparation procedure and increasesthe detection sensitivity. Applying this protocol to biologicalmuscle samples proves it to be a very sensitive, accurate, andprecise analytical tool.Supplementary key words muscle tissue • ion-pairing high-performance liquid chromatography • high-performance liquid chromatography-mass spectrometry
