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Background and purpose: Antibiotics are crucial in various scientific fields, including human and veterinary medicine, and are widely utilized medicinal compounds. Ciprofloxacin is a widely used antibiotic in the treatment of infections, most of which is excreted unmetabolized and finally enters water sources through the discharge of sewage and effluents. Even at low concentrations, ciprofloxacin in sewage and effluents can cause ecological harm and pose significant risks to human health. It is crucial to devise effective solutions for the removal of this antibiotic. Advanced Oxidation Processes (AOPs) are a promising method for the future removal of antibiotics, particularly in aquatic solutions, due to their potential to effectively remove ciprofloxacin. This study investigated the antibacterial and catalytic activity of synthesized magnetite nanoparticles (MNPs) (Fe3O4) in removing ciprofloxacin. Materials and methods: In this experimental research, Fe3O4 magnetic nanoparticles (MNPs) were prepared using the co-precipitation method. The nanoparticles were then examined for their physical and structural characteristics using scanning electron microscopy(SEM), X-ray powder diffraction (XRD), transmission electron microscopy (TEM), and BET technique. Fe3O4 magnetic nanoparticles (MNPs) were initially synthesized, and their properties were determined. We performed separate investigations to assess the effectiveness of various methods for removing ciprofloxacin. This included evaluating ultrasound alone (US), hydrogen peroxide alone, magnetite nanoparticles alone (MNPs), and combinations such as ultrasound with hydrogen peroxide, hydrogen peroxide with magnetite nanoparticles, and magnetite nanoparticles with ultrasound. All tests were conducted under the same conditions. The first step assessed the catalytic activity of synthesized nanoparticles for ciprofloxacin through the Fenton process. In this study, experimental parameters such as Fe3O4 catalyst dosage, initial pH of the solution, Hydrogen peroxide concentration, reaction time, and initial antibiotic concentration were investigated. In the second step, the antibacterial efficacy of synthesized nanoparticles against Escherichia coli and Staphylococcus aureus was determined via the Broth Macro dilution method. Results: The examination of the characteristics of synthesized nanoparticles showed that the average size of magnetite nanoparticles is approximately 20-30 nm. TEM results confirmed that the synthesized nanoparticles have a uniform size and structure. The combined process of MNPs/hydrogen peroxide/US exhibited the highest removal efficiency at 88.36%. This high efficiency can be attributed to the direct effect of the reaction between Fenton agents and ultrasonic waves, leading to the generation of numerous hydroxyl radicals. The maximum removal of ciprofloxacin was achieved in 0.5 g/L catalyst, pH=3, five mM hydrogen peroxide concentration, 550 W ultrasonic power, and 60 min reaction time. For Escherichia coli and Staphylococcus aureus, the Minimum Inhibitory and Minimum bactericidal concentrations were 3.125, 6.25, and 1.56, 3.125 μg/mL, respectively. Conclusion: Synthesized iron magnetite nanoparticles have high antibacterial properties against bacterial strains and many catalytic properties in the Sono Fenton process to remove ciprofloxacin, so this process can be an effective method for eliminating hospital and pharmaceutical wastewater.