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In the recent era of research and development, silver nanoparticles (AgNPs) are the most attractive nanomaterial in the development of nanomedicines due to their specific and distinctive physiochemical properties. Nanotechnology deals with the Nanoparticles having a size of 1-100 nm in one dimension used significantly concerning medical chemistry, atomic physics, and all other known fields. These particles can be prepared easily by different physical, chemical, and biological approaches. During the formation of AgNPs, the reduction of Ag+++to Agº occurs in presence of reducing and stabilizing agents which are naturally present in fungi, bacteria, yeast, and plants. In this study, we fabricate the AgNPs using the pineapple peels extract and characterization and optimization were carried out using the UV spectrophotometer, FTIR, and SEM techniques. Silver nanoparticles of Ananas comosus peel extract show the absorbance peak at around 410 nm analyzed by UV-Vis spectroscopy and their spherical shape and size (50 nm to 80 nm) was confirmed by the SEM analysis. FTIR analysis broke the point that the carbonyl group of reducing sugar stabilized the silver nanoparticles. These nanoparticles showed significant antimicrobial activity against Staphylococcus aureus and Escherichia coli. In the future researcher can explore the biomedical properties for the preparation of biosensors using AgNPs and their application in medicine by using the herbal medium.