Liquid Chromatography-Mass Spectrometry-Based In Vitro Metabolic Profiling Reveals Altered Enzyme Expressions in Eicosanoid Metabolism.

摘要: BACKGROUND Eicosanoids are metabolites of arachidonic acid that rapidly biosynthesized and degraded during inflammation, their metabolic changes reveal altered enzyme expression following drug treatment. We developed an eicosanoid profiling method evaluated on METHODS Simultaneous quantitative 32 eicosanoids in liver S9 fractions obtained from rabbits with carrageenan-induced inflammation was performed validated by liquid chromatography-mass spectrometry coupled to anion-exchange solid-phase purification. RESULTS The limit quantification for the devised ranged 0.5 20.0 ng/mg protein, calibration linearity achieved (R²>0.99). precision (% CV) accuracy bias) 4.7 10.3% 88.4 110.9%, respectively, overall recoveries 58.0 105.3%. Our then applied showed epitestosterone treatment reduced levels all were generated cyclooxygenases lipoxygenases. CONCLUSIONS Quantitative combined vitro assays may be useful evaluating affected drugs metabolism.