title: Identification of lipid derivatives in Hep G2 cells.
abstract: Metabolism of polyunsaturated fatty acids results in biosynthesis of mediators with different physiological effects. These metabolites include prostaglandins, prostacyclins, isoprostanes and others that are important signalling molecules and regulate a variety of physiological and pathophysiological processes including inflammation. Prostaglandins and isoprostanes are produced by either non-enzymatic lipid peroxidation or by enzyme-induced peroxidation (cyclooxygenases and lipoxygenases). They are used as biomarkers of oxidative stress. The aim of our study was to assess the effect of eicosapentaenoic acid (EPA) supplementation with added benzo(a)pyrene (BaP) on HepG2 cells by using a UHPLC/MS-TOF method. This rapid and simple method was developed for the identification, separation and quantification of 8-iPGF3α, PGF3α, 8-isoPGF2α and 5-iPF2α in cultured cells. The UHPLC/MS-TOF method was validated. The calculated limit of detection was in the range of 0.16-0.50 ng/mL, precision (% RSD): 1.2-2.1% and recoveries better than 88%. This method empowered qualitative and quantitative analysis of the selected individual prostaglandins derived from arachidonic acid and eicosapentaenoic acid from cell extracts.
authors.affiliation: Jagiellonian University, Medical College, Faculty of Pharmacy, Department of Radioligands, Kraków, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Gdula-Argasińska
authors.affiliation: Jagiellonian University, Faculty of Chemistry, Laboratory of Forensic Chemistry, Kraków, Poland.
authors.forename: Aneta
authors.initials: A
authors.lastname: Garbacik
authors.affiliation: Jagiellonian University, Medical College, Faculty of Pharmacy, Department of Radioligands, Kraków, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Tyszka-Czochara
authors.affiliation: Jagiellonian University, Faculty of Chemistry, Laboratory of Forensic Chemistry, Kraków, Poland.
authors.forename: Michał
authors.initials: M
authors.lastname: Woźniakiewicz
authors.affiliation: Jagiellonian University, Medical College, Faculty of Pharmacy, Department of Food Chemistry and Nutrition, Kraków, Poland.
authors.forename: Paweł
authors.initials: P
authors.lastname: Paśko
authors.affiliation: Jagiellonian University, Medical College, Faculty of Medicine, Infections Disease Clinic, Kraków, Poland.
authors.forename: Jacek
authors.initials: J
authors.lastname: Czepiel
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D015118:Eicosapentaenoic Acid / Q000494:pharmacology; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D007249:Inflammation / Q000378:metabolism* / Q000473:pathology; D028421:Isoprostanes / Q000302:isolation & purification / Q000378:metabolism; D057230:Limit of Detection; D015227:Lipid Peroxidation / Q000187:drug effects*; D018384:Oxidative Stress / Q000187:drug effects*; D011453:Prostaglandins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D015415:Biomarkers; D005231:Fatty Acids, Unsaturated; D028421:Isoprostanes; D011453:Prostaglandins; D015118:Eicosapentaenoic Acid