The toxicological implications of the presence of pharmaceuticals and personal care products (PPCPs) in the aquatic environment remain largely unknown. Acute toxicity tests have generally failed to detect the subtle action elicited by those compounds at environmentally relevant concentrations and they have often overlooked the fact that toxicity can be influenced by additive and synergistic effects. The aim of this study was to further assess the cytotoxicity of different pharmaceuticals and synthetic musks as well as their mixtures on the rainbow trout liver cell line RTL-W1. Eleven pharmaceuticals from different therapeutic classes (anti-inflammatory drugs, serotonin re-uptake inhibitors and lipid regulators) and five synthetic musks from the two major groups (nitro- and polycyclic musks) were selected for the study. Two fluorescent dyes were used to monitor cell viability. Among the tested compounds, estimated EC50s (effective concentration causing 50% decline of cell viability) denoted that polycyclic musks (7-25 microM) followed by anti-depressives (7-50 microM) showed the highest potential to induce cytotoxicity, whereas lipid regulators (20-380 microM), anti-inflammatory drugs (160-260 microM) and nitromusks (100-240 microM) had the lowest toxicity. Within a given therapeutic class, combined toxicity of mixtures was additive, following in most cases the concentration addition concept. However, the combined toxicity was higher than additive for those mixtures that included one compound from each class (i.e. dissimilar mixtures). Overall, this study shows that in the aquatic environment, toxicity of PPCPs on non-target organisms may occur at concentrations lower than expected due to synergistic effects between the different toxicants.