Based on the strong anti-proliferative activity of compound 8 on the four hepatocarcinoma cell lines, its ability to induced apoptosis was investigated. This was evaluated through measurement of Annexin V-FITC and PI staining of HepG2 cells, 24 hours after incubation with compound followed by flow cytometry for detection of these markers. The results (Table , Figure ) showed that HepG2 cells treated with madecassic acid (2) at 1x GI50 exhibit a higher proportion of cells in early and late apoptosis when compared with the untreated control (9.00 and 4.29% respectively) and that increasing the MA concentration to 3x GI50 increased the proportion of cells in both early and late apoptosis further . This result is in accordance with the reported induction of apoptosis by MA when colon cancer cells are treated with this compound. Compound 8 at 1 x GI50 resulted in induction of early and late apoptosis of 10.6% and 4.92% respectively along with a strong necrotic signal of 13.32%, whilst at 3 x GI50 early and late apoptosis values were lower than the untreated control, whilst necrosis was still increased (13.81%) Because cysteinyl aspartate specific proteinase-3 (caspase-3) plays an important role in the apoptotic pathway, the capacity of madecassic acid (2) and compound 8 to induce caspase 3 activity was measured at 1, 6, and 24 hours post treatment. The results in Table demonstrate that compound 2 caused a significant increase in caspase 3 activity in the HepG2 cells at 6 h post treatment, and at 3