In the present study, green silver nanoparticles (Ag 2 ONPs) were prepared from aqueous and ethanolic leaves extract of Rhamnus virgata in a facile, green, cost-effective, and eco-friendly way. The color changes from light brown to brownish black determined the synthesis of Ag 2 ONPs (Aq) and Ag 2 ONPs (Et). The phytofabrication of Ag 2 ONPs was confirmed using various spectroscopic and microscopic techniques: energy-dispersive X-ray spectroscopy, dynamic light scattering, ultraviolet-visible spectroscopy, Fouriertransform infrared, X-ray powder diffraction, Raman, scanning electron microscopy, and transmission electron microscopy. Detailed in vitro biological activities determined significant biopotentials for Ag 2 ONPs. The Ag 2 ONPs (Aq) and Ag 2 ONPs (Et) were investigated for anticancer potential against HUH-7 (IC 50 : 9.075 μg/ml for Ag 2 O (Aq) and 25.66 μg/ml for Ag 2 O (Et)) and HepG2 (IC 50 : 25.18 μg/ml for Ag 2 O (Aq) and IC 50 : 27.74 μg/ml for Ag 2 O (Aq)) cell lines. Concentration-dependent cytotoxicity was performed against brineshrimps (IC 50 : 36.04 μg/ml for Ag 2 O (Aq) and 28.82 μg/ml for Ag 2 O (Et)) and Leishmanial parasite (amastigotes and promastigotes). Disc-diffusion method revealed significant antimicrobial activities. In addition, significant enzyme inhibitory activity and antiradical potentials were studied. The hemocompatible nature of Ag 2 ONPs (Aq) and Ag 2 ONPs (Et) was revealed using biocompatibility tests. In conclusion, the green Ag 2 ONPs (Aq) and Ag 2 ONPs (Et) are nontoxic and biocompatible and has shown significant biological activities. We further encourage in vivo studies to ensure biosafety and biocompatibility, so that they can be effectively utilized in nano-pharmaceutical industries.