This study used a hydrothermal (autoclave) method to successfully prepare iron oxide nanoparticles (Fe2O3-NPs) by combining (Fe2NO3) salt with chitosan extract. In the antibacterial process, the IO-NPs are crucial in getting rid of germs and creating a healthy environment. IO-NPs were investigated using XRD spectra ,FE-SEM, UV-visible and, PL. According to the XRD results, Fe2O3 NPs have a face center cubic (FCC) structure and a crystal size of 10 to 25 nm. The diffraction peaks for Fe2O3 NPs can be assigned with good crystal quality. Furthermore, the FE-SEM results showed that the Fe2O3-NPs had grain size of 66 to 88 nm. The energy band gap, according to UV-visible spectra, is 4.2 eV. The inhibition of zones, which were 20 to 26 mm for gram-positive bacteria (Staphylococcus aureus), was used to determine the antibacterial qualities of Fe2O3-NPs.