title: LC-MS/MS for the simultaneous analysis of arachidonic acid and 32 related metabolites in human plasma: Basal plasma concentrations and aspirin-induced changes of eicosanoids.
abstract: Eicosanoids play an important role in various biological responses and can be used as biomarkers for specific diseases. Therefore, we developed a highly selective, sensitive, and robust liquid chromatography-tandem mass spectrometric method to measure arachidonic acid and its 32 metabolites in human plasma. Sample preparation involved solid phase extraction, which efficiently removed sources of interference present in human plasma. Chromatographic separation was performed using a Luna C(8)-column with 0.5mM ammonium formate buffer and acetonitrile as the mobile phase under gradient conditions. Detection was performed using tandem mass spectrometry equipped with an electrospray ionization interface in negative ion mode. The matrix did not affect the reproducibility and reliability of the assay. All analytes showed good linearity over the investigated concentration range (r>0.997). The validated lower limit of quantitation for the analytes ranged from 10 to 400pg/mL. Intra- and inter-day precision (RDS%) over the concentration ranges for all eicosanoids were within 16.8%, and accuracy ranged between 88.1 and 108.2%. This assay was suitable for the determination of basal plasma levels of eicosanoids and the evaluation of effect of aspirin on eicosanoid plasma levels in healthy subjects.
authors.affiliation: Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Busan, Republic of Korea.
authors.forename: Dhananjay D
authors.initials: DD
authors.lastname: Shinde
authors.forename: Kwon-Bok
authors.initials: KB
authors.lastname: Kim
authors.forename: Kyung-Suk
authors.initials: KS
authors.lastname: Oh
authors.forename: Nagi
authors.initials: N
authors.lastname: Abdalla
authors.forename: Kwang-Hyeon
authors.initials: KH
authors.lastname: Liu
authors.forename: Soo Kyung
authors.initials: SK
authors.lastname: Bae
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Shon
authors.forename: Ho-Sook
authors.initials: HS
authors.lastname: Kim
authors.forename: Dong-Hyun
authors.initials: DH
authors.lastname: Kim
authors.forename: Jae Gook
authors.initials: JG
authors.lastname: Shin
mesh_terms: D016718:Arachidonic Acid / Q000097:blood* / Q000737:chemistry / Q000378:metabolism; D001241:Aspirin / Q000494:pharmacology*; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D008297:Male; D055442:Metabolome / Q000187:drug effects; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D016718:Arachidonic Acid; D001241:Aspirin