Vol. 7, Issue 2, Part C (2025)

This study aimed to isolate high-purity lutein from Trigonella foenum-graecum (methi) leaves and evaluate its protective effects against oxidative DNA damage. Lutein was extracted via open-column chromatography on neutral alumina using hexane and dichloromethane:methanol (1:1, v/v), and its purity and structure were confirmed by UV-Vis spectroscopy, thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and liquid chromatography-mass spectrometry (LC-MS). Bioefficacy was assessed by exposing human lymphocytes and calf thymus DNA to oxidative stress induced by hydrogen peroxide (H₂O₂), ultraviolet (UV) radiation, and Fe²⁺/ascorbate (Fe:AA) systems, with DNA integrity analyzed using agarose gel electrophoresis and the diphenylamine assay. UV-Vis analysis showed a λmax at 445.5 nm, matching authentic lutein (446 nm), while TLC and HPLC confirmed ~99% purity, and LC-MS produced a major ion at m/z 551.48 ([M−H₂O + H]⁺), consistent with lutein’s molecular signature. Methi lutein (10-30 µM) markedly reduced DNA fragmentation, outperforming butylated hydroxyanisole (BHA) and α-tocopherol (each at 400 µM); at 30 µM, it completely inhibited H₂O₂-induced DNA degradation and preserved DNA integrity under UV exposure (345 nm) for up to 60 min at 20 µM. In the Fe:AA model, it achieved a 42% reduction in DNA fragmentation, exceeding the ~50% protection observed with BHA and α-tocopherol. This research provides a promising foundation for the development of natural antioxidant-based therapeutics, addressing the urgent need for effective strategies to mitigate oxidative genotoxicity with greater efficacy and safety.