Amoxicillin (AMX) and Ciprofloxacin (CIP) are antibiotics commonly used in human medicine with high environmental toxicity and poor biodegradability. They have been found in various hospital effluents and groundwater, and their environmental impact is still not fully understood. In this work, we investigated the possibility of treating model wastewaters containing the antibiotics AMX and CIP using ozonation, with the addition of H₂O₂ under various conditions, including different pH values, H₂O₂, and ozone dosages. The quantification of and treatment efficacy for antibiotic removal were determined via solid phase extraction followed by chromatographic separation by liquid chromatography coupled with tandem triple quadrupole mass spectrometry (LC/MS/MS). This analytical system is quite efficient for the detection of all major antibiotic classes, even if they are present at very low concentrations. The efficiency of ozonation was determined by measuring the TOC (Total Organic Carbon) changes after ozonation of the model wastewater and by measuring the concentration of the two antibiotics. In a sequential activated sludge process of ozone-treated model wastewater, almost complete TOC removal and an overwhelming decrease in antibiotic concentrations (up to 99%) were observed. Ozonation resulted in complete removal of AMX and CIP in less than 30 and 120 min, respectively. The results of this work indicate that ozonation could be a suitable pretreatment method to reduce the toxicity of contaminants (AMX and CIP) and improve the biodegradability of hospital wastewater.