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Today is an age of Green Technology. The green syntheses of nanoparticles have proven to be one of the greatest achievements in the field of Bionanotechnology 1. Off course, biosynthesized nanoparticles have been potential used for wide variety of appliances 2-3. In this regard we have paid emphasized to fabricate nanoparticles via cost-effective, eco-friendly, less hazardous and less-steps procedures 4-5. The phytosynthesis includes surfactant, capping agent, and stabilizing agents as phytochemicals. Considering these advantages, in the present work Zinc Oxide nanoparticles (ZnO NPs) was synthesized by using Cleome Simplicifolia. As-phytosynthesized material was further confirmed by electron spectroscopy techniques such as XRD, FTIR, UV-DRS, EDS, Raman, XPS, SEM, TEM and BET to know its structure, chemical state, size, shape, surface area and porosity. Diffuse reflectance spectroscopy (DRS) was used to evaluate an optical band gap energy of ZnO NPs which was found to be 3.15 eV, however TEM reveals nanoparticles sizes about 11-17 nm. Further, interestingly an antibacterial activity of ZnO NPs was performed during lunar eclipse against human pathogenic bacteria gram-positive (Staphylococcus aureus) and gram-negative