Zn-doped CuFe2O4 nanoparticles (NPs) were eco-friendly synthesized using plant extract. These nanoparticles were characterized by X-ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy and thermal gravimetric analysis (TGA). SEM image showed spherical NPs with size range less than 30 nm. In the EDS diagram, the elements of zinc, copper, iron, and oxygen are shown. The cytotoxicity and anticancer properties of Zn-doped CuFe2O4 NPs were evaluated on macrophage normal cells and A549 lung cancer cells. The cytotoxic effects of Zn-doped CuFe2O4 and CuFe2O4 NPs on A549 cancer cell lines were analyzed. The Zn-doped CuFe2O4 and CuFe2O4 NPs demonstrated IC50 values 95.8 and 278.4 µg/mL on A549 cancer cell, respectively. Additionally, Zn-doped CuFe2O4 and CuFe2O4 NPs had IC80 values of 8.31 and 16.1 µg/mL on A549 cancer cell, respectively. Notably, doping Zn on CuFe2O4 NPs displayed better cytotoxic effects on A549 cancer cells compared with the CuFe2O4 NPs alone. Also spinel nanocrystals of Zn-doped CuFe2O4 (~ 13 nm) had a minimum toxicity (CC50 = 136.6 µg/mL) on macrophages J774 Cell Line.