In this study, the toxic effects of binary mixtures of Pharmaceutical Active Compounds (PhACs) that are observed in the effluents from a wide range sewage treatment plants and surface water bodies were investigated using the bioluminescent bacterium Aliivibrio fischeri (Microtox® test). The selected chemicals were the nonsteroidal anti-inflammatory drug (NSAID) diclofenac sodium [DCF] and the anti-epileptic carbamazepine [CBZ]. In addition, caffeine [CFF], a psychoactive stimulant of the central nervous system, was also included in the study. Binary combinations were prepared at a predefined ratio that corresponded to the individual IC50 values of the investigated compounds (equitoxic ratio). The experimental results were compared with those obtained using the two most frequently used predictive models in aquatic toxicology: the Concentration Addition (CA) and Independent Action (IA) models. The results indicated that both models predict the observed mixture toxicity of the DCF-CBZ and DCF-CFF mixtures quite well. However, in the case of CFF-CBZ, both models slightly overestimated the experimental results, suggesting the presence of a potential antagonistic effect. The application of the Combination Index (CI) method, which allows us to identify and quantify the nature of the interactions between the chemicals present in a mixture (synergistic, additive or antagonistic effects), confirmed the additive behaviour of the DCF-CBZ and DCF-CFF combinations and the slightly antagonistic effect observed for the binary mixture of CFF-CBZ.

Di Nica, V., Villa, S., & Finizio, A. (2016). Experimental and predicted toxicity of binary combinations of diclofenac sodium, carbamazepine and caffeine to aliivibrio fischeri. ENVIRONMENTAL ENGINEERING AND MANAGEMENT JOURNAL, 15(9), 1971-1980.

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