A solvent-free method for spiking terrestrial algae (Desmococcus spp.) with pyrene for use in bioassays

Two methods for spiking terrestrial algae (Desmococcus spp., used as a foodstuff in bioassays with Collembola) with pyrene were tested; a "traditional" method that used a carrier solution of pyrene in acetone, and a solvent-free method developed using the principles of partition driven administration (PDA). The PDA method used a pre-spiked, highly-contaminated C18 disk as the source for pyrene, suspended and rotated in an aqueous algal suspension. The pyrene partitioned from the C18 disk into the aqueous phase, and a concentration of 18.4 +/- 0.7 microg pyrene g(-1)dwt. algae (mean +/- sd) was achieved after 18 h exposure by this method, with good reproducibility also observed for lower concentrations achieved over shorter exposure periods. The acetone-spiked algae lost much of its cell integrity and a significant amount of pigment, while cell integrity was maintained after 120 h spiking using the PDA method. Results from a short bioassay with Orchesella cincta (Collembola, Insecta) showed that animals fed acetone-spiked algae had slower growth rates and higher concentrations of pyrene metabolites than animals fed PDA-spiked algae. It is speculated that this was the result of the poor quality of the acetone-spiked food, and that the difference in food quality between treatments stimulated changes in body composition that may have affected the production of pyrene metabolites. The PDA spiking method is expected to be suitable for introducing a range of persistent organic pollutants into other types of sample matrix.