<p>The potential negative impact of antibiotics in effluents and drinking water poses health risks to the living biota. Therefore, attention has been focused on the removal of antibiotics from wastewater. A UV light in the presence of H₂O₂-based advanced oxidation process (AOP) was employed to degrade and detoxify moxifloxacin (MOX). The efficiency was evaluated based on degradation and reduction in cytotoxicity and mutagenicity. Box–Behnken design (BBD) was used for the RSM study to optimize and validate the degradation efficiency of moxifloxacin MOX. The UV irradiation alone degraded the drug up to 51.95%, while UV/H₂O₂ achieved a degradation of 62.9%. In addition, COD removal rates of 48% and 56% were observed for the UV irradiation alone and UV/H₂O₂ treatments, respectively. HPLC and GC–MS were used to identify the end-products, while FTIR was employed to analyze the functional groups. The cytotoxicity of the moxifloxacin samples was monitored using <i>A. Cepa</i>, Brine Shrimp, and hemolytic bioassays, while the Ames test evaluated mutagenicity before and after treatment for the first time in the current research. The bioassays showed that the toxicity of the drug solution was reduced significantly after treatment. Results revealed that UV radiation along with H₂O₂ is an effective tool for drug degradation which could be implemented for the effective degradation of wastewater containing antibiotics, particularly moxifloxacin.</p> Graphical abstract <p></p>