PARENT SESSION
HA6 General and multipurpose analysis
9:00 AM to 1:30 PM, Thursday, 10 May 2001
Session Chair: P. Sorensen
Room 6

(468) Immunoassays of crab urine to monitor exposure to PAH.

Fillmann, Gilberto^1,2,3, Watson, Giles³, Francioni, Eleine⁴, Readman, James², Depledge, Michael³, ^{1 2 3 4}

ABSTRACT- During the last decade immunoassays have been developed to detect selected pollutants in water and sediment/soil samples. Several commercial kits are now available for this purpose and offer rapid screening at costs much lower than by conventional techniques. Improvements in immunoassay generally relate to sensitivity and specificity. Recently, however, researchers have begun to apply the techniques to the analysis of biological media. Pollutants and their metabolites become concentrated in the tissues and body fluids of all biota thereby providing a measure of exposure to biologically "available" contaminants. The main objective of our work was to extend this approach to the analysis of crab urine to provide a non-destructive ecotoxicological tool (urine can be repeatedly sampled without damage to the organism) for monitoring PAHs and their metabolites in these organisms. Following one week of acclimation, crabs (Carcinus maenas) were exposed in filtered seawater (salinity 34 ppt) to the PAHs phenanthrene and pyrene at nominal concentrations ranging from 20-200 g L-1 in separate experiments. Concentrations in the tanks were monitored using GC-MS. Following 48 hours exposure, urine samples (50-200 L) were taken from crabs by opening the antennal gland opercula and allowing urine to flow out for collection. The samples were split (volumetrically) for analysis by immunoassay (enzyme-linked immunosorbent assay (ELISA) with antibodies attached to magnetic particles) and UV-fluorescence spectrophotometry. Following corrections using relative response factors (RRFs), high correlations between the immunoassay and spectrofluorometric analyses were found (r2=0.75 for phenanthrene and r2=086 for pyrene). Results demonstrate the potential for the use of this technique in environmental assessments and work is currently continuing to investigate relationships between exposure and the extent of adverse biological effects.

Key words: Immunoassay, PAHs, Urine, Crab