title: Liquid Chromatography-Mass Spectrometry-Based In Vitro Metabolic Profiling Reveals Altered Enzyme Expressions in Eicosanoid Metabolism.
abstract: 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.
authors.affiliation: Materials and Life Science Research Division, Korea Institute of Science and Technology, Seoul, Korea.|Department of Chemistry, Yonsei University, Seoul, Korea.
authors.forename: Su Hyeon
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Cardiovascular Center, Korea University Guro Hospital, Seoul, Korea.
authors.forename: Eung Ju
authors.initials: EJ
authors.lastname: Kim
authors.affiliation: Materials and Life Science Research Division, Korea Institute of Science and Technology, Seoul, Korea.
authors.forename: Dong Hyoung
authors.initials: DH
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Yonsei University, Seoul, Korea.
authors.forename: Won Yong
authors.initials: WY
authors.lastname: Lee
authors.affiliation: Materials and Life Science Research Division, Korea Institute of Science and Technology, Seoul, Korea.
authors.forename: Bong Chul
authors.initials: BC
authors.lastname: Chung
authors.affiliation: Cardiovascular Center, Korea University Guro Hospital, Seoul, Korea. mdhsseo@korea.ac.kr.
authors.forename: Hong Seog
authors.initials: HS
authors.lastname: Seo
authors.affiliation: Materials and Life Science Research Division, Korea Institute of Science and Technology, Seoul, Korea. mh_choi@kist.re.kr.
authors.forename: Man Ho
authors.initials: MH
authors.lastname: Choi
mesh_terms: D000818:Animals; D002351:Carrageenan / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D016207:Cytokines / Q000097:blood; D004195:Disease Models, Animal; D015777:Eicosanoids / Q000032:analysis* / Q000378:metabolism / Q000592:standards; D007249:Inflammation / Q000209:etiology / Q000378:metabolism; D008297:Male; D011817:Rabbits; D012015:Reference Standards; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000592:standards
chemical_list: D016207:Cytokines; D015777:Eicosanoids; D002351:Carrageenan
keywords: Arachidonic acid; Eicosanoids; Epitestosterone; Liquid chromatography-mass spectrometry; Liver S9 fraction