Pharmaceuticals are emerging contaminants in the environment and are a ubiquitous presence in rivers downstream of wastewater treatment plant outfalls. Questions remain about the persistence of pharmaceuticals in rivers, and the uptake and bioconcentration of pharmaceuticals by aquatic plants. Our study took place in the Yarrowee/Leigh/Barwon River system in southeastern Australia. We quantified the concentrations of five pharmaceuticals (carbamazepine, primidone, propranolol, tramadol, and venlafaxine) in surface water at five sites along a 144-km stretch of river, downstream of the presumed primary point source (a wastewater treatment plant outfall). We quantified pharmaceuticals in the leaves of two aquatic plant species (Phragmites australis and Vallisneria australis) sampled at each site, and calculated bioconcentration factors. All five pharmaceuticals were detected in surface waters, and the highest detected concentration exceeded 500 ng.