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Here, we report a simple, eco-friendly and inexpensive approach for the synthesis of zinc oxide nanoparticles (ZnO NPs) using Coptidis Rhizoma. The ZnO NPs were characterized by UV–visible absorption spectroscopy, FTIR, SEM-EDX, TGA, TEM, SAED and XRD. TEM images confirmed the presence of spherical and rod shaped ZnO NPs in the range of 2.90–25.20 nm. Green synthesized ZnO NP_S exhibited moderate antibacterial activity against Gram-positive and Gram-negative bacteria and excellent DPPH free radical scavenging activity. Synthesized ZnO NPs had no toxic effects on the RAW 264.7 cell line.