Internal concentrations link external exposure to the potential effect, as they reflect what the organisms actually take up and experience physiologically. In this study, we investigated whether frequently detected risk-driving substances in water were found in the exposed organisms and if they are classified the same based on the whole body internal concentrations. Field gammarids were collected upstream and downstream of ten wastewater treatment plants in mixed land use catchments. The sampling was conducted in autumn and winter, during low flow conditions when diffuse agricultural input was reduced. The field study was complemented with laboratory and flume experiments to determine the bioaccumulation potentials of selected substances. For 32 substances, apparent bioaccumulation factors in gammarids were determined for the first time. With a sensitive multiresidue method based on online-solid phase extraction followed by liquid chromatography coupled to high resolution mass spectrometry, we detected 63 (semi-) polar organic substances in the field gammarids, showing higher concentrations downstream than upstream. Interestingly, neonicotinoids, which are particularly toxic toward invertebrates, were frequently detected and were further determined as major contributors to the toxic pressure based on the toxic unit approach integrating internal concentration and toxic potency. The total toxic pressure based on internal concentrations was substantially higher compared to when external concentrations were used. Thus, internal concentrations may add more value to the current environmental risk assessment that is typically based solely on external exposure.

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