title: Development of a semi-automated LC/MS/MS method for the simultaneous quantitation of 14,15-epoxyeicosatrienoic acid, 14,15-dihydroxyeicosatrienoic acid, leukotoxin and leukotoxin diol in human plasma as biomarkers of soluble epoxide hydrolase activity in vivo.
abstract: Substrates and products of soluble epoxide hydrolase (sEH) such as 14,15-epoxyeicosatrienoic acid (14,15-EET), 14,15-dihydroxyeicosatrienoic acid (14,15-DHET), leukotoxin, and leukotoxin diol are potential biomarkers for assessing sEH activity in clinical trial subjects. To quantify them, we have developed and validated a semi-automated and relatively high-throughput assay in a 96-well plate format using liquid chromatography-mass spectrometry. 14,15-EET, 14,15-DHET, leukotoxin and leukotoxin diol, as well as their deuterium labeled internal standards were extracted from human plasma by liquid-liquid extraction using ethyl acetate. The four analytes were separated from other endogenous lipid isomers using liquid chromatography coupled with tandem mass spectrometry. The method was validated over a concentration range of 0.05-50 ng/mL. The validation results show that the method is precise, accurate and well-suited for analysis of clinical samples. The turn-around rate of the assay is approximately 200 samples per day.
authors.affiliation: GlaxoSmithKline Pharmaceuticals, 709 Swedeland Road, King of Prussia, PA 19406, USA. penny.x.zhu@gsk.com
authors.forename: Penny
authors.initials: P
authors.lastname: Zhu
authors.forename: Brian
authors.initials: B
authors.lastname: Peck
authors.forename: Hermes
authors.initials: H
authors.lastname: Licea-Perez
authors.forename: James F
authors.initials: JF
authors.lastname: Callahan
authors.forename: Catherine
authors.initials: C
authors.lastname: Booth-Genthe
mesh_terms: D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000097:blood; D015415:Biomarkers / Q000097:blood; D002853:Chromatography, Liquid; D004851:Epoxide Hydrolases / Q000097:blood / Q000378:metabolism*; D005260:Female; D006801:Humans; D008041:Linoleic Acids / Q000097:blood*; D008297:Male; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013229:Stearic Acids / Q000097:blood*; D053719:Tandem Mass Spectrometry
chemical_list: C107016:14,15-dihydroxyeicosatrienoic acid; C120038:9,10-dihydroxy-12-octadecenoic acid; D015415:Biomarkers; D008041:Linoleic Acids; D013229:Stearic Acids; C051208:9,10-epoxy-12-octadecenoic acid; C046782:14,15-epoxy-5,8,11-eicosatrienoic acid; D004851:Epoxide Hydrolases; D015126:8,11,14-Eicosatrienoic Acid