Background: Cancer is one of the major causes of death worldwide. It starts when cells grow out of control and crowd out normal cells. Current drug therapy possesses many severe side effects which need safe and effective agent that have the potential to resolve the disease condition. Aim: Evaluation of Cytotoxic potential effect of Catechin on the Cancer cells. Materials and Method: Procurement of the pure Catechin Powder from YUCCA Enterprises, Mumbai. Cytotoxic effects of Catechin were evaluated with the Standard marketed anticancer drug Methotrexate. Different models like In vitro hemolytic test, Brine Shrimp lethality assay and Allium cepa (onion) model was used to investigate the cytotoxic effect and the result was estimated. To check the significance of data, Statistical tests were performed: using ANOVA test and Tuckey’s test. Results and discussion: The effect of Catechin at different concentrations was observed on the cancer cells. Using models like In vitro Hemolytic test the % Hemolysis was observed at the concentration of 150 µg/mL, in Brine Shrimp lethality assay % mortality was calculated at the concentration of 300µg/mL at the interval of 6 hours and in Allium cepa model, the length and numbers of roots were counted. From this study we can conclude that natural phenol Catechin has the potential cytotoxic effect on the Cancer cells. Conclusion: Present research work emphasizes that the Catechin is having beneficial cytotoxic potential and may have therapeutic potential as anti-cancer agents.