Dietary intake of phytochemicals having antioxidant activity is associated with a lower risk of mortality from many diseases. Therefore, the aim of this study was to determine the free radical scavenging, cytotoxic and hemolytic activities of leaves of Acacia niloticaby using various methods. The results of the present study revealed that ethanol extract was the most effective and IC50value was found to be 53.6μg mL−1for Vero cell lines and 28.9μg mL−1for Hela cell lines in cytotoxicity assays. The zone of color retention was 14.2 mm in -carotene bleaching assay, which was as significant as positive control, butylated hydroxy toluene. None of the tested extracts possessed any hemolytic activity against rat and human erythrocytes revealing their cytotoxic mechanism and non-toxicity. Thus, only the ethanol extract could be considered as a potential source of anticancer and antioxidant compounds. Further phytochemical studies will be performed for specification of the biologically active principles.