Phenolic Composition, Chemometric Analysis, and Multifunctional Biological Activities of Organ-Specific Extracts From Endemic Hedysarum erythroleucum

Hedysarum erythroleucum is a plant species native to Türkiye and represents a potential source of bioactive compounds. This study investigated the phytochemical composition and biological activities of methanolic extracts prepared from the leaves, stems, and roots. LC-MS/MS analysis identified 53 phenolic compounds and revealed clear differences in metabolite distribution among plant organs. Quinic acid was the predominant phenolic acid, particularly in the leaf extract. Flavonoid glycosides such as isoquercitrin, astragalin, and rutin were also abundant in the aerial parts of the plant. The antioxidant potential of the extracts was evaluated using DPPH, FRAP, total phenolic content, and total flavonoid content assays. Among the tested plant parts, the leaf extract exhibited the strongest antioxidant activity, with the lowest DPPH IC50 value, the highest reducing power, and the highest total phenolic and flavonoid contents. These findings indicate that the leaves contain high levels of antioxidant compounds. Enzyme inhibition assays showed that the leaf extract exhibited the highest inhibitory activity against acetylcholinesterase, butyrylcholinesterase, and α-glycosidase. Its activity was comparable to that of the reference inhibitors under the experimental conditions. Antiproliferative activity was assessed against lung and breast cancer cell lines. All extracts inhibited cancer cell growth, with the highest sensitivity observed in H1650 and MDA-MB-231 cells. Tumor selectivity indices suggested moderate selectivity, especially for root extracts. LDH cytotoxicity analysis indicated limited membrane damage, suggesting that the extracts primarily exert cytostatic effects by suppressing cell proliferation rather than inducing extensive cell death. Multivariate analyses revealed organ-specific metabolite patterns and associations between phenolic composition and the measured biological activities. Overall, the findings indicate that H. erythroleucum, particularly the leaf extract, contains a high content of phenolic compounds and exhibits promising in vitro antioxidant, enzyme inhibitory, and antiproliferative activities. These findings provide a basis for future phytochemical investigations, mechanistic studies, compound isolation, and in vivo validation. © 2026 Royal Society of New Zealand Te Apārangi.