A solid-phase extraction method combined with liquid chromatography tandem mass spectrometry was developed to analyze simultaneously prostaglandins (PGE2, PGE3), prostacyclins (6-keto-PGF1α, Δ17-6-keto-PGF1α), resolvins (RvD1, RvD2) and leukotriene (LTB4) released into cL-15 medium by salmon liver cells. The optimal concentrations of different internal standards, for determining the analytical performance parameters, were selected by means of a uniform shell design. The limit of detection, quantification and recovery for the seven released pro- and anti-inflammatory biomarkers into cL-15 medium ranged from 0.3-1.0 ng/mL, 0.5-2.0 ng/mL and 83-127% respectively. The validated method was used to investigate the effect of polyunsaturated fatty acids (PUFA) on the production of prostaglandins, prostacyclins, resolvins and leukotriene by salmon liver cells. Statistically significant increases in the concentration of some eicosanoids were observed after adding arachidonic acid (PGE2, 6-keto-PGF1α and Δ17-6-keto-PGF1α) and eicosapentaenoic acid (PGE2, 6-keto-PGF1α, Δ17-6-keto-PGF1α and LTB4). Neither linoleic nor docosahexaenoic acid affected the production of both arachidonic acid or docosahexaenoic acid derived metabolites. Although RvD1 and RvD2 were not detected, there was some indication that the production of RvD3 and RvD4 was preferred over RvD1 and RvD2 after exposing the cells to different PUFA.

Keywords: Prostaglandins; Prostacyclins; Resolvins; Leukotriene; Solid Phase Extraction; Liquid Chromatography Mass Spectrometry; Cell Cultures; Atlantic salmon (Salmo salar).