The study focused on the medicinal properties of citrus fruits and their ability to synthesize silver nanoparticles. As the resistance against the modern antibiotic agents is on increase, finding new and effective natural antibiotic agents is the need of the modern era. Similarly, bio-synthesis of nanoparticles is also being encouraged for eco-friendly reasons. Due to remarkable medicinal and industrial applications of silver nanoparticles (AgNPs), citrus fruit juice is used to reduce silver ions for the green synthesis of AgNPs. Phytochemical analysis revealed the presence of various constituents which impart antibacterial property to citrus fruits, analyzed against four pathogenic bacteria. Also, citrus fruit juice exhibits radical scavenging activity because of these constituents. Further, the AgNPs synthesized using citrus fruits were characterized using Ultra-Violet Visible (UV-VIS) spectroscopy, Fourier Transform Infrared (FTIR) spectroscopy and Transmission Electron Microscopy (TEM) to study the shape and size of the AgNPs. Anticancer activity of AgNPs was also evaluated against Colo-205 cell lines and found to inhibit 37.9% growth of cell lines at the concentration of 10 μg/ml. Hence, synthesized AgNPs can be used effectively against cancer cell lines in combination with other anti-cancer agents.