In the current decade, the use of plant extracts for the green preparation of metal nanoparticles has garnered increasing attention due to their eco-friendliness, cost-effectiveness, and sustainability. In the current study, silver nanoparticles (AgNPs) were synthesized using the ethanolic extract of Circaea lutetiana for the first time. Thetotal flavonoid content (TFC) and total phenolic content (TPC)of the extract were analyzed by spectrophotometric methods. Fourier transform infrared (FT-IR) spectroscopy was employed to determine the functional groups involved in both the reduction and stabilization processes of AgNPs. The formation and optical properties of AgNPs were confirmed by Ultraviolet–Visible (UV–Vis) spectroscopy. The greenlysynthesized AgNPs were characterized by FT-IR, UV–Vis, X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), dynamic light scattering (DLS) and zeta potential analyses, transmission electron microscopy (TEM), and scanning electron microscopy (SEM). The results confirmed that the AgNPs were spherical in shape with an average size of approximately 3.8 nm and showed a good crystalline nature. Additionally, the AgNPs exhibited significant antimicrobial activity against both Gram-positive and Gram-negative bacteria, demonstrating their potential as green antimicrobial agents.