Optimization of the Ames/salmonella mutagenicity assay for use with extracts of aquatic sediments

Non-mutagenic components interfered with the ability of the standard Ames/salmonella assay to detect mutagenicity in extracts of contaminated Great Lakes sediments. The use of gel permeation chromatography (GPC) to remove these macromolecules from methylene chloride extracts prior to Ames testing enhanced the likelihood of transfer of mutagenic components into dimethyl sulf oxide (the assay solvent). Therefore, to optimize the assay's sensitivity we pre-treated sediment extracts using GPC and increased metabolic activity through the use of a 30% S9 mix. Increasing the level of Aroclor 1254-induced rat liver S9, typically used to metabolically activate promutagens, had the additional beneficial effect of reducing the cytotoxicity of the extracts. As applied in this study, the Ames assay can serve as a sensitive test for screening the mutagenic potential of large numbers of uncharacterized sediment extracts.