One new cycloartane-type triterpenoid, named 24-epimarkhacanasin C (1), together with four known flavonoids, apigenin (2), luteolin (3), apigenin 7-O-β-D-glucopyranoside (4), and luteolin 7-O-β-Dglucopyranoside (5), was isolated by various chromatographic methods from the ethyl acetate extract of Markhamia stipulata var. canaense leaves. Their structures were elucidated by IR, UV, HR-ESI-MS, and NMR experiments. All compounds were reported for the first time from this species. Their cytotoxicities were evaluated against human breast cancer cell line (MCF-7) by SRB assay. The isolated compounds 1, 3, and 4 exhibited significant cytotoxic activity (IC50 ranged from 53.38 to 68.32 μg/mL), while compounds 2 and 5 did not show activity.