In the present study, green silver nanoparticles (Ag₂ONPs) 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 Ag₂ONPs_(Aq) and Ag₂ONPs_(Et). The phytofabrication of Ag₂ONPs was confirmed using various spectroscopic and microscopic techniques: energy‐dispersive X‐ray spectroscopy, dynamic light scattering, ultraviolet–visible spectroscopy, Fourier‐transform infrared, X‐ray powder diffraction, Raman, scanning electron microscopy, and transmission electron microscopy. Detailed in vitro biological activities determined significant biopotentials for Ag₂ONPs. The Ag₂ONPs_(Aq) and Ag₂ONPs_(Et) were investigated for anticancer potential against HUH‐7 (IC₅₀: 9.075 μg/ml for Ag₂O_(Aq) and 25.66 μg/ml for Ag₂O_(Et)) and HepG2 (IC₅₀: 25.18 μg/ml for Ag₂O_(Aq) and IC₅₀: 27.74 μg/ml for Ag₂O_(Aq)) cell lines. Concentration‐dependent cytotoxicity was performed against brine‐shrimps (IC₅₀: 36.04 μg/ml for Ag₂O_(Aq) and 28.82 μg/ml for Ag₂O_(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 Ag₂ONPs_(Aq) and Ag₂ONPs_(Et) was revealed using biocompatibility tests. In conclusion, the green Ag₂ONPs_(Aq) and Ag₂ONPs_(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.