| Summary: | <i>Polygonum hydropiperoides</i> Michx. is an Asian native plant species that is also widely distributed in the Americas. Despite its traditional uses, <i>P. hydropiperoides</i> is scarcely scientifically exploited. This study aimed to chemically characterize and investigate the antioxidant and antibacterial activities of hexane (HE-Ph), ethyl acetate (EAE-Ph), and ethanolic (EE-Ph) extracts from aerial parts of <i>P. hydropiperoides</i>. The chemical characterization was performed through HPLC-DAD-ESI/MS<sup>n</sup>. The antioxidant activity was assessed by the phosphomolybdenum reducing power, nitric oxide inhibition, and the β-carotene bleaching assays. The antibacterial activity was determined by the minimal inhibitory concentration (MIC) and the minimal bactericidal concentration followed by the classification of the antibacterial effect. Chemical characterization revealed the expressive presence of phenolic acids and flavonoids in EAE-Ph. An increased antioxidant capacity was revealed in EAE-Ph. Regarding antibacterial activity, EAE-Ph showed weak to moderate property against 13 strains tested with MIC values ranging from 625 to 5000 µg/mL, with bactericidal or bacteriostatic effects. Glucogallin and gallic acid stand out as the most relevant bioactive compounds. These results suggest that <i>P. hydropiperoides</i> is a natural source of active substances, supporting this species’ traditional use.
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