Objective: Screening of preliminary phytochemicals, evaluation of in vitro antioxidant and in vitro anticancer activities of Simarouba glauca leaf extracts on T-24 Bladder cancer cell line. Materials and Methods: Herbal extraction was carried out by Soxhlet method using chloroform, ethylacetate, methanol, ethanol, aqueous and hydroalcohol. Phytochemical investigation was done using biochemical tests. Total phenolic content was estimated by Folin-Ciocalteu reagent (FCR) method. Antioxidant potential of leaf extracts was analyzed by Ferric ion reducing antioxidant power (FRAP) assay, Phosphomolybdenum (PM) assay and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. In vitro anticancer activity on T-24 bladder cancer cell line was assessed by MTT assay. Statistical analysis used: Statistical analysis of data was performed by analysis of variance (one-way ANOVA) and level of statistical significance between groups was carried out using GraphPad Prism version 5.0 for Windows (GraphPad Software, San Diego, CA, USA). Results: Phytochemical analysis revealed the presence of rich secondary metabolite present in all the solvent extracts. Hydroalcoholic extract showed highest presence of phenolic content (92.38±0.29 mg/g) GAE. Ethanol and methanol extract showed highest antioxidant capacity in DPPH, FRAP and PM assay as compared to the other extracts based on the test performed. The results confirmed that ethanol extract significantly (p<0.05) inhibited T-24 cell line with IC50 value (533.55±25.02 μg/mL) as compared to standard drug doxorubicin (0.16μM/mL). Conclusions: The results of the present findings strengthen the potential property of Simarouba glauca as a resource for the discovery of novel antioxidant and anticancer agents.