An efficient one-step scheme for the purification of major xanthophyll carotenoids from lettuce, and assessment of their comparative anticancer potential

The foremost problem in carotenoid research is the excessive cost and difficulty of maintaining pure carotenoid compounds. This work presents an economical, efficient, and simplified one-step scheme for the purification of four major xanthophyll carotenoids from lettuce by utilizing preparative thin layer chromatography on Hyflo-Super-Cel: MgO (Heavy): calcium sulfate hemihydrate (9:9:2 w/w) based adsorbent. The mobile phase of acetone: hexane (1:1) provided the perfect separation of major xanthophylls, resulting in 95-96% purity after just single-step separation, with no interference from chlorophylls or other minor carotenoids. The identity of carotenoids was confirmed by absorption spectroscopy, chemical tests and APCI⁺-MS/MS. The proposed scheme can be used to isolate the carotenoids at the analytical and preparative scale. In Anticancer studies, among four xanthophylls, 9-Z-neoxanthin was found most potent for reduction of cell viability of cervical (HeLa) and lung Cancer (A549) cells, with IC₅₀ values of 3.8 and 7.5 μM, respectively.