The use of spectrofluorimetry for monitoring the bioaccumulation and the biotransformation of polycyclic aromatic hydrocarbons in Daphnia magna

A rapid and direct spectrofluorimetric method was tested in order to monitor the bioaccumulation of PAHs in Daphnia magna in a control media. After exposure to Water containing benzo[a]pyrene or fluoranthene, daphnids were put in solvent, sonicated, and filtered. The fluorescence spectrum observed in the filtrate was recorded. Results were compared to HPLC measurements of the same pools of organisms. In fluoranthene experiments, the fluorescence peak of the daphnid extract spectrum was linearly related to the PAH content as measured with HPLC. In benzo[a]pyrene experiments, other fluorescent compounds progressively appeared in the sample. They were assumed to be metabolites. A linear regression involving fluorescence intensities at two different wavelengths was necessary for a satisfactory correlation with HPLC measurements, A water extraction was performed to isolate metabolites, None or very few fluoranthene metabolites were isolated in the aqueous phase, whereas increasing ben:a[a]pyrene metabolites were observed while the exposure time increased.