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KMID : 1100720160360040342
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Annals of Laboratory Medicine
2016 Volume.36 No. 4 p.342 ~ p.352
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Liquid Chromatography-Mass Spectrometry-Based In Vitro Metabolic Profiling Reveals Altered Enzyme Expressions in Eicosanoid Metabolism
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Lee Su-Hyeon
Kim Eung-Ju
Lee Dong-Hyoung
Lee Won-Yong
Chung Bong-Chul
Seo Hong-Seog
Choi Man-Ho
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Abstract
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Background: Eicosanoids are metabolites of arachidonic acid that are rapidly biosynthesized and degraded during inflammation, and their metabolic changes reveal altered enzyme expression following drug treatment. We developed an eicosanoid profiling method and evaluated their changes on drug treatment.
Methods: Simultaneous quantitative profiling of 32 eicosanoids in liver S9 fractions obtained from rabbits with carrageenan-induced inflammation was performed and validated by liquid chromatography-mass spectrometry coupled to anion-exchange solid-phase purification.
Results: The limit of quantification for the devised method ranged from 0.5 to 20.0 ng/mg protein, and calibration linearity was achieved (R2>0.99). The precision (% CV) and accuracy (% bias) ranged from 4.7 to 10.3% and 88.4 to 110.9%, respectively, and overall recoveries ranged from 58.0 to 105.3%. Our method was then applied and showed that epitestosterone treatment reduced the levels of all eicosanoids that were generated by cyclooxygenases and lipoxygenases.
Conclusions: Quantitative eicosanoid profiling combined with in vitro metabolic assays may be useful for evaluating metabolic changes affected by drugs during eicosanoid metabolism.
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KEYWORD
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Eicosanoids, Arachidonic acid, Liquid chromatography-mass spectrometry, Liver S9 fraction, Epitestosterone
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