Several Saharan plants, despite their abundance of natural compounds, have received little attention. In this study, the chemical composition of polar extracts of Tourneuxia variifolia Coss. (Asteraceae), an endemic species to Algerian Sahara, was investigated and their anticancer activity was evaluated in vitro. The phytoconstituents of both ethyl acetate (EtOAc) and n-butanol (n-BuOH) extracts were screened using LC/MS-MS technique. The anticancer activity of the above extracts was measured against human cervical adenocarcinoma (HeLa) cell line. The LC/MS-MS analyses results revealed that twenty-seven phytochemicals in EtOAc extract and twenty-three in n-BuOH extract were identified and quantified from which isoquercetin and astragalin were the most present. Moreover; the EtOAc extract was found to have a strong anticancer activity (IC50: 46.797 ± 0.060 µg/mL). These findings identified T. variifolia as a potential plant exhibiting anticancer properties.