In this present work, Fe3O4 nanoparticles were obtained by a one-part Co-precipitation method. Secondly, a simple solvothermal method was employed to synthesize Chitosan (CS) coated Fe3O4 nanoparticles. Chitosan was used as a surface modification agent. The surface of Chitosan-coated Fe3O4 nanoparticles was conjugated with Folic Acid (FA). Various properties of the resultant products were performed by characterization studies. The structure and surface morphology of as-prepared nanoparticles were characterized by X-ray diffraction (XRD) and Scanning electron microscopy (SEM). The physical parameter such as strain and the crystallite size was evaluated for all the reflection peaks of the samples by using Williamson–Hall (W-H plot) method. Determination of the chemical component was marked by Fourier transform infrared spectroscopy (FTIR) and Energy-dispersive X-ray spectroscopy (EDAX) analyses. UV- Vis experiment was carried out to record optical absorbance and the bandgap energy of the nanoparticles was determined by Tauc’s plot. Thermogravimetric analysis (TGA) was conducted to recognize the thermal stability of the magnetic nanoparticles and endothermic peaks were observed from the first derivative of the TGA curve (DTG curve).