A COMPARISON OF HPLC WITH FLUORESCENCE DETECTION AND FIXED WAVELENGTH FLUORESCENCE METHODS FOR THE DETERMINATION OF POLYCYCLIC AROMATIC HYDROCARBON METABOLITES IN FISH BILE

Abstract

Fish exposed to polycyclic aromatic hydrocarbons (PAHs) accumulate only a small amount of PAH in their tissues. Hydrophobic PAHs are readily transformed into hydrophilic metabolites and excreted in bile. Bile samples of five fish species were collected from the Baltic Sea and analyzed using two methods: high-performance liquid chromatography (HPLC) with fluorescence detection and a fixed wavelength fluorescence method. With the HPLC method PAH compounds were quantitated after hydrolysis, and with the fluorescence method without enzyme-assisted hydrolysis. In HPLC analysis the major metabolite in all fish species, 1-hydroxypyrene, was the only metabolite detected in perch and salmon samples. 1-Hydroxyphenanthrene was detected in a few flounder and in most eelpouts. The correlation between the two methods for pyrene-type metabolites was good (r² = 0.773).

Keywords:

• bile
• fish
• fixed fluorescence
• high-performance liquid chromatography method comparison
• polycyclic aromatic hydrocarbon metabolite

Acknowledgments

This work was partly supported by a grant from the European Union on Biological Effects of Environmental Pollution in Marine Coastal Ecosystems, http://beep.lptc.u-bordeaux.fr (EVK3-CT-2000-00025) and partly by the Maj and Tor Nessling Foundation (Grant nos. 2000210 and 2001133). The bile samples from flounder, eelpout, cod, and some of the perch were supplied by Katja Broeg (Alfred Wegener Institutes for Polar and Marine Research, Department of Ecotoxicology, Bremerhaven, Germany), Janina Barsiene (Institute of Ecology, Vilnius, Lithuania), Doris Schiedek and Sabine Schnell (Institut für Ostseeforschung Warnemünde, Rostock, Germany), and Jens Gercken (Institute of Applied Ecology, Neu Broderstorf, Germany). Salmon samples were collected by Timo Myllylä and some of the perch samples by Karl Sundman (Finnish Game and Fisheries Research Institute, Helsinki, Finland). Roy Siddall checked English language in the manuscript.

Notes

^a Waters Corporation, Milford, MA, USA.

^b Vydac 201TP54, Grace Vydac, Columbia, MD, USA.