Journal
JOURNAL OF LIPID RESEARCH
Volume 48, Issue 1, Pages 252-259Publisher
ELSEVIER
DOI: 10.1194/jlr.D600037-JLR200
Keywords
deuterium-labeled stearic acid; deuterium-labeled oleic acid; deuterium-labeled palmitic acid; high-performance liquid chromatography-electrospray ionization-; mass spectrometry; electrospray ionization negative mode; quantification of plasma levels
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Imbalanced fatty acid metabolism contributes significantly to the increased incidence of metabolic disorders. Isotope-labeled fatty acids (H-2, C-13) provide efficient means to trace fatty acid metabolism in vivo. This study reports a new and rapid method for the quantification of deuterium-labeled fatty acids in plasma by HPLC-MS. The sample preparation protocol developed required only hydrolysis, neutralization, and quenching steps followed by high-performance liquid chromatography-electrospray ionization-mass spectrometry analysis in negative ion mode using single ion monitoring. Deuterium-labeled stearic acid (d7-C18:0) was synthesized to reduce matrix interference observed with d5 analog, which improved the limit of detection (LOD) significantly, depending on the products analyzed. Linearity > 0.999 between the LOD(100nM) and 30 mu M, accuracy > 90%, precision > 88%, and adequate recovery in the dynamic range were obtained for d7-C18:0 and d7-oleic acid (C18:1). Upon oral dosing of d7-C18:0 in rats, the parent compound and its desaturation and beta-oxidation products, d7-C18:1 and d7-C16:0, were circulating with a maximal concentration ranging from 0.6 to 2.2 mu M, with significant levels of d7-fatty acids detected for up to 72h. -Gagne, S., S. Crane, Z. Huang, C. S. Li, K. P. Bateman, and J-F. Levesque. Rapid measurement of deuterium-labeled long-chain fatty acids in plasma byHPLC-ESI-MS.
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