Metabolomic study on ostracods exposed to environmentally relevant concentrations of five pharmaceuticals selected via a novel approach

Pharmaceuticals (PhACs) are increasingly detected in aquatic ecosystems, yet their effects on biota remain largely unknown. The environmentally relevant concentrations of many PhACs may not result in individual-level responses, like mortality or growth inhibition, traditional toxicity endpoints. However, this doesn't imply the absence of negative effects on biota. Metabolomics offers a more sensitive approach, detecting responses at molecular and cellular levels and providing mechanistic understanding of adverse effects. We evaluated bioaccumulation and metabolic alterations in a benthic ostracod, Heterocypris incongruens, exposed to a mixture of five PhACs (carbamazepine, tiapride, tolperisone, propranolol and amlodipine) at environmentally relevant concentrations for 7 days using liquid chromatography coupled with mass spectrometry. The selection of PhACs was based, among other factors, on risk quotient values determined using toxicological data available in the literature and concentrations of PhACs quantified in our previous research in the sediments of the Odra River estuary. This represents a novel approach to PhACs selection for metabolomic studies that considers strictly quantitative data. Amlodipine and tolperisone exhibited the highest bioaccumulation. Significant impacts were observed in Alanine, aspartate and glutamate metabolism, Starch and sucrose metabolism, Arginine biosynthesis, Histidine metabolism, Tryptophan metabolism, Glycerophospholipid metabolism, and Glutathione metabolism pathways. Most of the below-individual-level responses were likely nonspecific and related to dysregulation in energy metabolism and oxidative stress response. Additionally, some pharmaceutical-specific responses were also observed. Therefore, untargeted metabolomics can be used to detect metabolic changes resulting from environmentally relevant concentrations of PhACs in aquatic ecosystems and to understand their underlying mechanism.