Metoprolol (MTP) is a compound of concern, considered as an emerging contaminant due to its high consumption, pseudopersistence and potential ecotoxicity. Activated sludge batch experiments were performed to evaluate the biological transformation of MTP and the formation of transformation products under different treatment conditions. Total MTP removal was obtained in aerobic conditions, and the formation of MTP known metabolites (metoprolol acid (MTPA), α-hydroxymetoprolol (α-HMTP) and O-desmethylmetoprolol (O-DMTP)) and unknown transformation products (TPs) was investigated. The three known metabolites and two new TPs generated along the experiments were identified by liquid chromatography coupled to high resolution mass spectrometry. For the two new TPs plausible structures were proposed based on the tentative identification. MTPA had the major ratio formation for the TPs identified along the experiments (up to 40% of initial MTP concentration after 96 h treatment) and its persistence through biological treatment was proven. Ecotoxicity studies using Vibrio fischeribioluminescent bacteria in an acute toxicity test showed that MTP and its known TPs are not toxic with the exception of o-DMTP. Finally, MTP and its TPs were monitored in a full scale membrane bioreactor and in a full scale conventional urban wastewater treatment plant (WWTP) and the results were compared with those obtained in batch experiments. α-HMTP was detected for the first time in a WWTP influent whereas MTPA was detected in influent and effluent WWTP samples at much higher levels (up to 100 folds higher) than MTP itself remarking its high persistence.