Metabolite Profile and Biological Activities of Zanthoxylum nemorale: A Metabolomic and Multivariate Statistical...

This study investigated the metabolite profile and biological activities of Zanthoxylum nemorale leaf and stem extracts obtained using hexane, ethyl acetate, and methanol by combining untargeted LC-HRMS metabolomics, multivariate statistical analysis, antioxidant, and antimicrobial assays. A total of 112 metabolites were putatively identified, including coumarins, lignans, flavonoids, alkaloids, phenolic acids, terpenoids, and fatty acids. Methanol extracts exhibited the strongest antioxidant activity, with IC₅₀ values of 53.12 ± 1.11 and 60.70 ± 0.28 µg mL^{- 1} for stem and leaf extracts, respectively, and showed the highest antibacterial activity against Staphylococcus aureus and Bacillus cereus. Chemometric analyses (PCA, PLS-DA, and hierarchical clustering) clearly discriminated the extracts according to plant organ and solvent polarity. Overall, the extracts were more effective against Gram-positive bacteria than Gram-negative bacteria and fungi. These findings highlight Z. nemorale as a promising source of antioxidant and antimicrobial phytochemicals and demonstrate the value of metabolomics for linking chemical composition with biological activity.