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Background: Wedang Uwuh is a traditional Javanese functional beverage with historical roots in the Mataram Sultanate. Despite its traditional prominence, the comprehensive metabolite profile and underlying biological mechanisms of wedang uwuh remain insufficiently characterized, necessitating a systematic evaluation of its chemical constituents and associated bioactivities. Objective: This study characterized the metabolite profile of Wedang Uwuh using advanced mass spectrometry and evaluated its antioxidant and α-glucosidase inhibitory activities. Methods: Aqueous extracts were analyzed via Liquid Chromatography-Mass Spectrometry (LC-MS). Antioxidant capacity was measured as Ascorbic Acid Equivalent Antioxidant Capacity (AEAC), while α-glucosidase inhibition was tested across a 15.625–1000 ppm gradient. Results: LC-MS identified a complex phytochemical landscape dominated by flavonoids, tannins, and benzopyrans, featuring key markers such as brazilin, hesperetin, and cinnamtannin B1. The aqueous powder extract demonstrated superior antioxidant capacity (94.64 ppm AEAC) compared to ethanolic and traditional preparations. Furthermore, all extracts exhibited potent, dose-dependent α-glucosidase inhibition, reaching peak suppression at 1000 ppm. Conclusion: These in vitro findings demonstrate that wedang uwuh extract exhibits a potential role in glucose regulation via α-glucosidase inhibition, thereby supporting further exploration in in vivo models. The synergistic high antioxidant capacity and enzymatic inhibition position this beverage as a promising standardized functional drink for managing oxidative stress and postprandial hyperglycemia.
