The study aimed to investigate the profile of alkaloids in two ethyl acetate soluble fractions, namely fractionsA and B from an ethanolic extract of Ficus septica leaves and cytotoxic effect on T47D breast cancer cells.Preparation of both fractions involved maceration of leaves with 70% (v/v) ethanol, filtration with Al2O3,precipitation with 0.1 N HCl, Mayer reagent, and 0.1 N NaOH, and also partition with ethyl acetate. Qualitativethin layer chromatography (TLC) was conducted to determine the profile of alkaloids in the two fractions,using alkaloid specific reagents such as Dragendorff, sodium nitrite, and Van Urk-Salkowski. Cytotoxic effectsof both fractions on T47D cells were evaluated using MTT assay with a concentration series of 1.56; 3.12; 6.25;12.5; 25 and 50 μg/mL. The TLC test showed that fractions A and B contained alkaloids with Rx values of 0.74and 0.80 for fraction A and 0.74, 0.84, 0.92 for fraction B with regard to yohimbine using the mobile phase ofn-buthanol:glacial acetic acid:distilled water (3:1:1 v/v/v). Moreover, an indole alkaloid was detected with Rxvalues of 0.80 and 0.84, respectively. Fractions A and B exhibited high cytotoxic effects on T47D cells with IC50values of 2.57 and 2.73 μg/mL, respectively. In conclusion, overall the results of this study showed that fractionsof Ficus septica contain alkaloids including indole alkaloid or its derivatives and possess a cytotoxic effect onT47D cells. This research supports the idea that alkaloids in F. septica have anticancer activity.