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.
Bibliographical noteFunding Information:
This study was supported in part by a grant from the Korea Institute of Science and Technology Institutional Program (Project No. 2E26110) and a grant from the Bio and Medical Technology Development Program (NRF-2013M3A9B6046413) through the Ministry of Science, ICT and Future Planning.
© The Korean Society for Laboratory Medicine.
- Arachidonic acid
- Liquid chromatography-mass spectrometry
- Liver S9 fraction
ASJC Scopus subject areas
- Biochemistry, medical
- Clinical Biochemistry