Phytochemical Analysis and Bioevaluation of Echinophora sibthorpiana Guss. (Apiaceae): Antioxidant Capacity, DNA Protection and In Vivo Genotoxic Safety

Echinophora sibthorpiana Guss. (Apiaceae) is an aromatic plant of considerable ethnobotanical significance, traditionally employed as a flavouring and preservative agent in food and valued for its medicinal properties across the Eastern Mediterranean region, yet its non-volatile phytochemistry and biosafety profile remain insufficiently characterized. In the present study, the n-butanol fraction of the 80% methanolic extract of the aerial parts of E. sibthorpiana was fractionated using chromatographic methods and the structures of the isolated compounds were elucidated by 1D and 2D-NMR spectroscopy and HR-ESI-MS. The distribution of Echinophora metabolites isolated by our group across six species in Türkiye was assessed by LC-qTOF-MS. Antioxidant capacity was evaluated by CUPRAC, FRAP, and TEAC assays; DNA-protective activity by the pBR322 plasmid model; genotoxic and antigenotoxic potential in the Drosophila melanogaster wing SMART assay. Four secondary metabolites, known as vicenin-2 (1), rutin (2), isoquercitrin (3) and betulalbuside A (4), were isolated and reported from E. sibthorpiana for the first time. Notably, the C-glycoside flavone vicenin-2 and the acyclic monoterpene glucoside betulalbuside A also represent the first isolation of these compounds from the genus Echinophora. LC-qTOF-MS profiling identified widely distributed flavonoid constituents together with more restricted metabolites that may have chemotaxonomic relevance. The n-BuOH fraction and the flavonol glycosides rutin and isoquercitrin exhibited the highest antioxidant and DNA-protective activities. None of the tested samples displayed genotoxic activity, while all showed antigenotoxic effects against ethyl methanesulfonate-induced DNA damage, with total spot frequency inhibition ranging from 47% to 83%. Overall, these findings provide a first comprehensive characterization of the non-volatile phytochemistry and biosafety profile of E. sibthorpiana and support the further investigation of its constituents as safe antioxidant and chemopreventive agents with potential nutraceutical applications.