In biomedical science, the environmentally friendly and economical production of metal nanoparticles is becoming increasingly significant. Zinc oxide nanoparticles (ZnO NPs) were biosynthesized in this study from Citrus limon extract, and their anticancer efficacy was evaluated utilizing several cytotoxicity assays in JURKAT cancer (Leukemia) cell lines. The morphological and structural characteristics of biogenic ZnO NPs were characterized. The synthesis was confirmed using well-known analytical techniques like UV-visible spectrophotometry, XRD, FTIR, and scanning electron microscopy. The stability of the produced NPs was also investigated. SEM and TEM were used to capture the production of cubical nanoparticles with an average size of 52nm. With a 50% inhibitory dosage of 10 g/mL, we could observe the dose-dependent rise in cytotoxicity of ZnO NPs in the JURKAT cell line.Using an MTT experiment, citrus-produced ZnNPs demonstrated lower vitality with an IC50 value of 152.54μg/ml when compared to control cells (p<0.05), indicating the anti-proliferating action of the nanoparticles. The amount of LDH leakage was dose-dependently increased by our eco-friendly CuNPs (IC50: 85.43±1.25μg/ml).However, it is urged that this research be expanded across various cancer models and that, in light of the results, in vivo validation be conducted.