High performance liquid chromatographic separation of fish biliary polynuclear aromatic hydrocarbon metabolites.

A modification of Krahn's procedure for the high performance liquid chromatographic (HLPC) separation of fish biliary polynuclear aromatic hydrocarbon (PAHs) metabolites was examined by using three C-18 columns in tandem, eluted with a mobile phase gradient of 100% water to 100% acetonitile in 240 min. By following the modified procedure, the bile of spot (Leiostomus xanthurus), Atlantic croaker (Micropogonius undulatus), weakfish (Cynoscion regalis), hogchoker (Trinectes maculatus) and oyster toadfish (Opsanus tau) from the PAH-polluted Southern Branch of the Elizabeth River, Virginia gave rise to large numbers of reasonably well resolved PAH metabolite peaks compared to the control Nansemond River fish. The separation of fish biliary PAH metabolites, without elaborate sample extractions (an important feature of Krahn's procedure), indicate the possibility of direct detection and quantification of critical metabolites by HPLC AND HPLC/mass spectrometry (HPLC/MS).