Due to great calling for medical and antimicrobial textiles based on eco-friendly natural agents, silver nanoparticles were prepared using herbal extract peppermint, tulsi, chamomile, sage and green tea at optimized conditions. Plant extracts are very cost effective, eco-friendly and thus can be economic alternatively for large-scale synthesis of nanoparticles. In this research firstly, the herbal extracts were prepared by aqueous method, secondly using of herbal extracts in synthesis of silver nanoparticles from aqueous solutions of silver nitrate (AgNO3). Synthesis of silver nanoparticles was characterized and confirmed by UV–visible spectroscopy and TEM. The herbal extracts and silver nanoparticles were applied in presence of eco-friendly finishing agent (glyoxal, aluminuim sulphate) on cotton fabrics which in turn characterized using scanning electron microscopy (SEM), (EDX) patterns, physical properties as tensile strength, water permeability, air permeability and roughness were also analyzed. Final characterization of treated cotton fabric against antimicrobial activity including gram negative strain, gram positive strains in addition to yeasts unicellular fungi . The results showed that cotton treated with AgNPs synthesis from herbal extracts had better antimicrobial activity.