Hedera helix rhizomatifera L. plant belongs to the family Araliaceae. It is a host of a wide variety of bioactive compounds (mainly triterpenoidal saponins and phenolic compounds) of several biological activities; including spasmolytic, antiinflammatory, antimicrobial and many other activities. It is widely marketed as a herbal cough remedy. The aim of the present study was the phytochemical investigation of the species rhizomatifera, in addition to the assessment of its cytotoxicity on target cancer cell lines. Our results indicated that H. helix spp. rhizomatifera manifested a significant cytotoxic activity against Human hepatocellular carcinoma cell line (HepG2) and human breast cancer cell line (MCF7) with IC50 reaching 1.9125 and 2.0823 μg/ml respectively, in comparison to 1.549 and 1.02 μg/ml respectively for Doxorubicin. The UPLC-ESI-MS/MS study revealed the presence of 24 compounds, 15 of which were tentatively identified as triterpenoidal saponins, flavonoids mainly kaempferol and quercetin derivatives, in addition to some phenolic acids.