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The accumulation of 17 human pharmaceuticals (HPs) was investigated in the muscle of three fish species characteristic of the "Rio de la Plata Basin" with different feeding habits and of relevance for human consumption: Megaleporinus obtusidens, Salminus brasiliensis, and Prochilodus lineatus. Fish were sampled in fall and spring from 8 localities distributed along 500 Km of the Uruguay River. Atenolol and carbamazepine were the most frequently detected HPs (>50%), but at concentrations always below 1 μg/kg wet weight (w/w). Hydrochlorothiazide, metoprolol, venlafaxine, propranolol, codeine, and the carbamazepine metabolite, 2-hydroxycarbamazepine, were accumulated at higher levels showing maximum concentrations between 1 and 10 μg/kg (w/w), but infrequently (<50%). The other HPs were always below 1 μg/kg (w/w) and at frequencies lower than 50%. Distinctive accumulation patterns were observed among species at different trophic levels. However, biomagnification trends were not identified for any compound. The highest number and concentration of HPs were found in M. obtusidens (omnivorous), followed by P. lineatus (detritivorous), and lastly S. brasiliensis (piscivorous). The most recurrent HPs (i.e. carbamazepine and atenolol) were present in all species, but others exclusively in one. Geographical variations were only found for carbamazepine and atenolol in M. obtusidens and P. lineatus, showing higher concentrations in localities closer to the Rio de la Plata estuary. Differences in the HPs concentrations among seasons were not identified. Acceptable daily intake and predicted no effect concentrations would indicate that measured muscle concentrations in fish from the Uruguay River do not pose a serious risk for human consumption nowadays. Further studies will be necessary for assessing the potential adverse effects on studied fish species., (Copyright © 2019 Elsevier Ltd. All rights reserved.)