Characterization of biliary metabolites of fluoranthene in the common sole (Solea solea)

Abstract

Fluoranthene is one of the most abundant polycyclic aromatic hydrocarbon (PAH) pollutants in the environment. Studies of the metabolism of PAHs have highlighted the importance of the gallbladder in concentrating xenobiotics in fish before excretion in feces. Analysis of bile metabolites can be considered useful for monitoring and assessing the exposure of fish to PAHs. Although the fate of several PAHs in marine organisms has been widely investigated, information is lacking regarding the metabolism of fluoranthene in fish. Therefore, we investigated the metabolic pathways of [14C]fluoranthene in the common sole (Solea solea) by identifying bile metabolites using electrospray ionization/mass spectrometry (ESI/MS) and nuclear magnetic resonance (NMR) spectroscopy. [14C]Fluoranthene was administered by intraperitoneal injection to 20 common soles. Groups of animals (n = 5) were killed 1, 2, 3, and 4 d postdosing, and gallbladders were excised for radioactivity counting and bile analysis. Biliary metabolites were separated and quantified by radio-high-performance liquid chromatography, and structure identification was performed by ESI/MS. Isomeric structures were confirmed by NMR analyses. At the end of the experiment, 12.2% of the administered radioactivity was recovered in bile. As expected, hydroxylation and glucuronidation were the predominant metabolic pathways. The 7-O-glucuronide-fluoranthene metabolite (representing 13.3% of total radioactivity found in bile), 8-O-glucuronide-fluoranthene (11.8%), trans-2,3-dihydro-3-hydroxy-2-O-glucuronide-fluoranthene (17.9%), and cis-2,3-dihydro-2-hydroxy-3-O-glucuronide-fluoranthene (13.9%) were the major metabolites observed in bile. Minor metabolites, such as trans-2,3-dihydro-2-hydroxy-3-O-glucuronide-fluoranthene (3.9%) and 2,3-di-O-glucuronide-fluoranthene (6.6%), also were identified. The 2,3-dihydrodiol-fluoranthene metabolite, which is found in bile conjugated to glucuronic acid, would be, after hydrolysis of the conjugates, a suitable biomarker of PAH pollution in the marine environment.