Afonso, Tiago BarrosBonifácio-Lopes, TeresaCosta, Eduardo M.Macedo, TiagoMoreira, JoanaOliveira, Juliana A. S. A.Pintado, Manuela2026-04-222026-04-222026-05-01e7280fbd-d093-45e5-a9ca-f4d46e754984http://hdl.handle.net/10400.14/57557Pterospartum tridentatum (L.) Willk. (carqueja) is widely used in traditional medicine and culinary practices in the Iberian Peninsula; however, most studies have focused on its flowers, while its stems remain largely unexplored, despite representing a significant proportion of the plant biomass. This study aimed to evaluate the potential of P. tridentatum stems as a source of bioactive compounds using different extraction methodologies. Aqueous, hydroethanolic, ultrasound-assisted extraction (UAE) and pressurized liquid extraction (PLE) were applied, and the resulting extracts were characterized in terms of their extraction yield, protein and carbohydrate content, phenolic composition, antioxidant capacity, antimicrobial activity, and cytotoxicity in HaCaT and Caco-2 cell lines. Phenolic profiling by LC-ESI-QqTOF-HRMS tentatively identified 37 compounds, mainly corresponding to flavonoid and isoflavonoid glycosides, with genistein derivatives representing the dominant constituents across all extracts. Although extraction yields differed among methods, phenolic profiles were broadly similar. UAE and PLE extracts showed slightly higher antioxidant activity, while antimicrobial activity was limited, with only moderate inhibition observed against Staphylococcus epidermidis and Malassezia furfur. Additionally, cytotoxicity assays indicated low toxicity. Overall, the results demonstrate that P. tridentatum stems represent a promising yet underutilized biomass source of phenolic compounds with antioxidant potential and low cytotoxicity under the tested in vitro conditions.engPterospartum tridentatumGenista tridentataBiomass valorizationGreen extractionPhenolic compoundsIsoflavonoidsAntioxidant activityCytotoxicityresearch article10.3390/foods15091461