Ibuprofen (IBU) has been considered as one of emergent pharmaceutical contaminants in environments due to its occurrences in natural water bodies. Some reports suggested that the IBU was biodegradable but details about biodegradation pathways and functional microbial community were still not fully clear. This study was aimed to assess IBU biodegradation using three different activated sludges (i.e. H: hospital, M: municipal, and D: distillery) with foci on surmising degradation pathway based on UPLC/Q-ToF/MS (ultra-performance liquid chromatography quadrupole time of flight mass spectrometry) analyses and concluding microbial community according to high-throughput sequencing for partial 16S rRNA gene. Results showed that some IBU (∼5 mg/L) was able to be degraded only by sludges H and M during 2–5 days incubation under aerobic conditions. From LC/MS analysis of degradation byproducts, two major ring-opening precursors were identified in extracted ion chromatogram data. Ring-opening degradation pathways leading to the formation of low-molecular-weight carboxylic acids were elucidated. Additionally, the principal coordinate analyses using Fast UniFrac method for the partial 16S rRNA gene showed the microbial communities in the three sludges were significantly different but typically with high proportion of sequences matched gene fragments from Proteobacteria phylum. Some sequences with first matches with previously reported IBU degraders (i.e. Nocardia sp., Sphingomonas sp., and Variovorax sp.) were only found in the sludges H and M which showed capability for IBU biodegradation. These results demonstrated some functional microbes in activated sludges from hospital and municipal WWTPs had potential to break down IBU into smaller molecules.