Comparative phytochemical analysis of Humulus scandens (Lour.) Merr. ethanol and aqueous extracts using UHPLC-MS/mS combined with feature-based molecular networking

Humulus scandens (Lour.) Merr. (HS) is a medicinal plant belonging to the Cannabaceae family and widely distributed in East Asia. Although it has been traditionally used for health-related purposes, its phytochemical profile remains incompletely characterized, particularly regarding how different extraction solvents influence bioactive compounds. Water and ethanol are the two most commonly used solvents in herbal medicine and food processing. Water extraction reflects traditional decoction practices, while ethanol extraction is widely used in modern studies due to its ability to extract a broader range of compounds. Therefore, comparing these two solvents is important for linking traditional use with modern functional food development. In this study, UHPLC-MS/MS combined with feature-based molecular networking was applied to compare ethanol and aqueous extracts of HS. A total of 164 compounds were annotated in the ethanol extract and 136 in the aqueous extract, with 51 compounds shared. Statistical analysis revealed that N-p-coumaroyltyramine, feruloyltyramine, and arginine were significantly enriched in the ethanol extract. Although the two extraction methods exhibited largely overlapping phytochemical profiles, ethanol extraction provided broader metabolite coverage and preferentially enriched these three discriminatory metabolites. These findings provide a basis for selecting appropriate extraction methods and support the development of HS-derived functional foods and nutraceuticals. Phytochemical profiles of HS ethanol and aqueous extracts were compared using UHPLC-MS/MS coupled with feature-based molecular networking.A total of 164 bioactive constituents were annotated in the ethanol extract and 136 in the aqueous extract, with 51 compounds common to both.Ethanol extraction resulted in a greater diversity of bioactive constituents compared with aqueous extraction.N-p-coumaroyltyramine, feruloyltyramine, and arginine were significantly enriched in the ethanol extract. Phytochemical profiles of HS ethanol and aqueous extracts were compared using UHPLC-MS/MS coupled with feature-based molecular networking. A total of 164 bioactive constituents were annotated in the ethanol extract and 136 in the aqueous extract, with 51 compounds common to both. Ethanol extraction resulted in a greater diversity of bioactive constituents compared with aqueous extraction. N-p-coumaroyltyramine, feruloyltyramine, and arginine were significantly enriched in the ethanol extract.