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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 879; no. 25; pp. 2487 - 2493
• Main Authors: Zhu, Penny, Peck, Brian, Licea-Perez, Hermes, Callahan, James F., Booth-Genthe, Catherine
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.09.2011; Elsevier
• Subjects: 8,11,14-Eicosatrienoic Acid - analogs & derivatives; 8,11,14-Eicosatrienoic Acid - blood; Analysis; Analytical, structural and metabolic biochemistry; Assaying; Biological and medical sciences; biomarkers; Biomarkers - blood; Chromatography; Chromatography, Liquid; clinical trials; deuterium; Dihydroxyeicosatrienoic acid (DHET); Dihydroxyoctadec-12(Z)-enoic acid (leukotoxin diol); Diols; epoxide hydrolase; Epoxide Hydrolases - blood; Epoxide Hydrolases - metabolism; Epoxyeicosatrienoic acid (EET); Epoxyoctadec-12(Z)-enoic acid (leukotoxin); ethyl acetate; Female; Format; Fundamental and applied biological sciences. Psychology; General pharmacology; Human; Human plasma; Humans; isomers; LC/MS; Linoleic Acids - blood; Liquid chromatography; liquid-liquid extraction; Male; Medical sciences; Pharmacology. Drug treatments; Quantitation; Reproducibility of Results; Sensitivity and Specificity; Soluble epoxide hydrolase (sEH); Stearic Acids - blood; Surgical implants; Tandem Mass Spectrometry; LC/MS; Human plasma; Dihydroxyeicosatrienoic acid (DHET); Dihydroxyoctadec-12(Z)-enoic acid (leukotoxin diol); Quantitation; Soluble epoxide hydrolase (sEH); Epoxyeicosatrienoic acid (EET); Epoxyoctadec-12(Z)-enoic acid (leukotoxin); Biological fluid; Chemical analysis; HPLC chromatography; Biological marker; Epoxide hydrolase; Blood plasma; Development; Quantitative analysis; Human; Automation; Diol; leukotoxin; Enzyme; Epoxyoctadec-12(Z)-enoic acid; Dihydroxyoctadec-12(Z)-enoic acid; In vivo; leukotoxin diol; Mass spectrometry MS/MS; Simultaneous measurement; Ether hydrolases; Hydrolases; Mass spectrometry
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2011.06.042

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.