Xanthophyll carotenoids, such as lutein and zeaxanthin, may provide potential health benefits in humans against chronic and degenerative diseases. We recently demonstrated cleavage of the xanthophylls lutein and zeaxanthin by carotene‐9’,10’‐monooxygenase (CMO2) at the 9’,10’ and 9,10 double bonds, producing 3‐OH‐β‐apo‐carotenoids. In the present study, we investigated the potential biological activity of the xanthophyll metabolite 3‐OH‐β‐apo‐10’‐carotenal (3‐OH‐ACL) in vitro. We found that incubation of 3‐OH‐ACL with BEAS‐2B, A549 and HepG2 cell lines dose‐ and time‐dependently decreased cell viability. Utilizing High Content Screening (HCS) analysis, we observed that 3‐OH‐ACL treatment significantly increased cytochrome C release and nuclear swelling and decreased mitochondrial potential in HepG2 cells. We further explored the effect of 3‐OH‐ACL on the LPS‐stimulated TNFα, MCP1 and IL‐6 expression in THP‐1 human monocyte/macrophage cells. 3‐OH‐ACL significantly inhibited LPS‐stimulated MCP1 and IL‐6 gene expression but at higher concentrations increased TNFα gene expression in THP‐1 cells. These data indicate that 3‐OH‐β‐apo‐10’‐carotenal may induce apoptotic pathways of cellular death.Grant Funding Source: R01CA104932