Photocatalytic degradation of antibiotic rifampicin (RIF) in aqueous solution, employing 100-nm sized anatase TiO2 nanoparticles (NPs) as a catalyst, under 365-nm light irradiation (0.28 mW cm�2) was investigated. The direct photolysis and adsorptive removal of the ansamycin antibiotic were found to be negligibly inefficient. The experimental data suggest that photocatalysis efficiently degrades RIF, and the effect of irradiation time, catalyst dosage, pH of medium, and temperature was elucidated. The optimum photocatalytic degradation occurred in the colloidal mixture of 30.38 μM RIF solution and 10 mg TiO2 NPs at pH 5.63. The activation energy was estimated to be 1.585 ± 0.079 kJ mol�1. The photocatalytic degradation kinetics of the antibiotic followed the Langmuir�Hinshelwood model with the rate constant of 0.016 ± 0.002 min�1, and the additional of a small amount of H2O2 accelerates the photocatalytic process. The degradation pathway was discussed based on vibrational spectroscopic data of RIF and its by-products. Overall, the findings in this study suggest that photocatalysis employing anatase TiO2 NPs under UV light irradiation is a promising approach for the degradation of antibiotics and other pharmaceuticals in wastewater treatment plants. Graphical abstract: (Figure presented.) © Akadémiai Kiadó, Budapest, Hungary 2024.