出版社：American Society for Biochemistry and Molecular Biology
摘要：Imbalanced fatty acid metabolism contributes significantly tothe increased incidence of metabolic disorders. Isotope-labeledfatty acids (2H, 13C) provide efficient means to trace fattyacid metabolism in vivo. This study reports a new and rapidmethod for the quantification of deuterium-labeled fatty acidsin plasma by HPLC-MS. The sample preparation protocol developedrequired only hydrolysis, neutralization, and quenching stepsfollowed by high-performance liquid chromatography-electrosprayionization-mass spectrometry analysis in negative ion mode usingsingle ion monitoring. Deuterium-labeled stearic acid (d7-C18:0)was synthesized to reduce matrix interference observed withd5 analog, which improved the limit of detection (LOD) significantly,depending on the products analyzed. Linearity > 0.999 betweenthe LOD (100 nM) and 30 µM, accuracy > 90%, precision> 88%, and adequate recovery in the dynamic range were obtainedfor d7-C18:0 and d7-oleic acid (C18:1). Upon oral dosing ofd7-C18:0 in rats, the parent compound and its desaturation andß-oxidation products, d7-C18:1 and d7-C16:0, werecirculating with a maximal concentration ranging from 0.6 to2.2 µM, with significant levels of d7-fatty acids detectedfor up to 72 h.Supplementary key words 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
Abbreviations: C16:0, palmitic acid; C16:1, palmitoleic acid; C18:0, stearic acid; C18:1, oleic acid; HPLC-ESI-MS, high-performance liquid chromatography-electrospray ionization-mass spectrometry; LOD, limit of detection; MRM, multiple reaction monitoring; SIM, single ion monitoring
