The study aims to determine the secondary metabolites of Hypericum androsaemum L. extracts by liquid chromatography-high resolution mass spectrometry (LC-HRMS), and investigate the antioxidant and cytotoxic activities of the plant. Cytotoxic activity was evaluated by MTT assay, and apoptosis induction abilities on human prostate adenocarcinoma (PC-3), and hepatocellular carcinoma (Hep G2) cell lines. Accordingly, major secondary metabolites were found as hederagenin (762 +/- 70.10 mu g/g) in the leaves dichloromethane (LD), herniarin (167 +/- 1.50 mu g/g) in fruit dichloromethane (FD), (-)-epicatechin (6538 +/- 235.36 mu g/g) in the leaves methanol (LM), (-)-epigallocatechin gallate (758 +/- 20.46 mu g/g) in the fruit methanol (FM), and caffeic acid (370 +/- 8.88 mu g/g) in the fruit water (FW), and (3313 +/- 79.51 mu g/g) in the leaves water (LW) extracts. LM exerted strong antioxidant activity in DPPH free (IC50 10.94 +/- 0.08 mu g/mL), and ABTS cation radicals scavenging (IC50 9.09 +/- 0.05 mu g/mL) activities. FM exhibited cytotoxic activity with IC50 values of 73.23 +/- 3.06 mu g/mL and 31.64 +/- 2.75 mu g/mL on PC-3 and Hep G2 cell lines, respectively. Being the richest extract in terms of quillaic acid (630 +/- 18.9 mu g/g), which is a well-known cytotoxic triterpenoid with proven apoptosis induction ability on different cells, FM extract showed apoptosis induction activity with 64.75% on PC-3 cells at 50 mu g/mL concentration. The study provides promising results about the potential of Hypericum androsaemum on cancer prevention.