Identification of lipid derivatives in Hep G2 cells.

Joanna Gdula-Argasińska; Aneta Garbacik; Małgorzata Tyszka-Czochara; Michał Woźniakiewicz; Paweł Paśko; Jacek Czepiel

doi:10.18388/abp.2013_2064

Joanna Gdula-Argasińska Jagiellonian University, Medical College, Faculty of Pharmacy, Department of Radioligands, Kraków, Poland.;
Aneta Garbacik Jagiellonian University, Faculty of Chemistry, Laboratory of Forensic Chemistry, Kraków, Poland.;
Małgorzata Tyszka-Czochara Jagiellonian University, Medical College, Faculty of Pharmacy, Department of Radioligands, Kraków, Poland.;
Michał Woźniakiewicz Jagiellonian University, Faculty of Chemistry, Laboratory of Forensic Chemistry, Kraków, Poland.;
Paweł Paśko Jagiellonian University, Medical College, Faculty of Pharmacy, Department of Food Chemistry and Nutrition, Kraków, Poland.;
Jacek Czepiel Jagiellonian University, Medical College, Faculty of Medicine, Infections Disease Clinic, Kraków, Poland.;

DOI: https://doi.org/10.18388/abp.2013_2064

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.