<p>The present study aimed to synthesize zinc ferrite nanoparticles (ZnFe₂O₄ NPs) using the sol-gel method and evaluate their antibacterial properties against <i>Escherichia coli</i> (<i>E. coli</i>) and <i>Staphylococcus aureus</i> (<i>S. aureus</i>), along with an assessment of their antioxidant activity. Characterization, including X-ray diffraction (XRD) analysis, confirmed that the nanoparticles exhibited a face-centered cubic (FCC) crystalline structure. Transmission electron microscopy (TEM) revealed that the nanoparticles were spherical in shape, with an estimated size ranging from approximately (~30–60 nm). The magnetic properties were investigated using a vibrating sample magnetometer (VSM), while Fourier-transform infrared spectroscopy (FTIR) identified the functional groups present. Notably, this study presents a novel approach by demonstrating, for the first time, the enhanced antibacterial efficacy of ZnFe₂O₄ nanoparticles under an alternating magnetic field (AMF), highlighting their unique DNA-mediated mechanism of action. Furthermore, the ZnFe₂O₄ NPs exhibited sustained antioxidant activity as measured by the DPPH assay. These combined antioxidant and AMF-enhanced antibacterial properties suggest promising pharmacological and biomedical applications for ZnFe₂O₄ NPs, particularly in the development of magnetically responsive therapeutic agents. Notably, this study presents a novel approach by demonstrating, for the first time, the enhanced antibacterial efficacy of ZnFe₂O₄ nanoparticles under an alternating magnetic field (AMF), highlighting their unique DNA-mediated mechanism of action. Furthermore, the ZnFe₂O₄ NPs exhibited sustained antioxidant activity as measured by the DPPH assay. These combined antioxidant and AMF-enhanced antibacterial properties suggest promising pharmacological and biomedical applications for ZnFe₂O₄ NPs, particularly in the development of magnetically responsive therapeutic agents.</p> Graphical Abstract <p></p>