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Abstract

A rapid method was developed for the simultaneous profiling of 29 free fatty acids in plasma using ultra-performance liquid chromatography/electrospray ionization tandem mass spectrometry (UPLC/ESI-MS/MS). Barium acetate was used as the cationization agent in the positive ion mode for sensitive multiple reaction monitoring (MRM) experiments. The cis- and trans-C18:1 and -C18:2 isomers were baseline-separated using two tandem reversed-phase C18 UPLC columns, while identification of two pairs of positional isomers of C18:3 and C20:3 required isomer-specific product ions, as the analytes were not chromatographically resolved. The assay linearity was greater than three orders of magnitude and correlation coefficients were >0.99; the limits of detections were typically less than 0.2 µM. The method was successfully applied to plasma free fatty acid profiling of samples from volunteers who participated in a randomized crossover study involving the administration of either placebo or fish oil capsules. The results clearly indicate the ability to measure the time profiles of the n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in plasma for the volunteers given fish oil capsules while the concentrations of the other free fatty acids and the total free fatty acid concentration in plasma remained virtually constant. Copyright © 2008 John Wiley & Sons, Ltd.