Aim And Objective: The aim of the present study was to synthesize and characterize Titanium Dioxide nanoparticles (TiO2 NP) using the leaf of Eucalyptus Globulus. To evaluate the shape and size of TiO2 NPs synthesized using the leaf of Eucalyptus globulus. Material And Methods: In 100ml distilled water, 1 g of Eucalyptus globulus was mixed and dissolved. The herbal formulation was heated for 15 minutes at 70ºC and filtered. Using an orbital shaker, this filtrate was homogeneously combined with 0.39mg of Titanium dioxide and then used to bio reduce titanium (Ti) ions to titanium dioxide nanoparticles (TiO2 NPs). The surface characterization of the titanium dioxide nanoparticles was assessed using UV–Visible absorbance spectroscopy, Scanning Electron Microscopy (SEM) and functional group evaluation using Fourier transform infrared spectroscopy (FTIR). Result: Scanning electron microscope results revealed that the synthesized Titanium dioxide nanoparticles were spherical in shape and their average size ranged from 20–70 nm. Conclusion: Titanium dioxide nanoparticles can be effectively extracted from Eucalyptus globulus when used as a reducing agent.