Zinc Ferrite nanoparticles were prepared by chemical coprecipitation method, using PEG as the capping agent. The synthesized materials were annealed at different temperatures such as 400, 500, 600, 700 and 800 °C and the structural characterization studies were conducted by X-Ray Diffraction (XRD), Fourier Transform Infrared Spectrometer (FTIR), UV-Visible Spectroscopy and Field Emission Scanning Electron Microscopy (FESEM). On heat treatment of the as prepared samples, crystallite size was observed to increase from 5 nm-22 nm. XRD pattern confirmed the single crystalline phase of developed particles. It was observed that the particle size and X-ray density of the annealed ZnFe 2 O 4 nanoparticles linearly increased with increase in annealing temperature and decrease in lattice parameters. The calculated micro strain values were found to be decreasing with increasing annealing temperatures. The nanoparticle size was confirmed using SEM measurements. Furthermore FTIR mapping confirmed the presence of ZnFe 2 O 4 nanoparticles. The calculated band gap (Eg) of zinc ferrite nanoparticles decreases with increase in temperature. The antibacterial activity of ZnFe 2 O 4 nanoparticles was studied against gram negative E-coli pathogenic bacteria. DPPH assay was used to determine the antioxidant activity of the compound. In-vitro anticancer activity was screened by MTT assay against MCF-7 (Human breast adenocarcinoma) cell and A549 (Lung Cancer) cell line. It was observed that the increase in particle size and annealing temperature influence the inhibition rate of these bio properties. The investigation points out that ZnFe 2 O 4 nanoparticle possess less viability with increase in concentration towards MCF-7 cell than A549 cell.