Tarenaya aculeata is traditionally used in Brazil to treat inflammation and pain-related conditions.This study aimed to investigate the chemical composition, anti-inflammatory potential, and genotoxicity of a freeze-dried stem decoction of T. aculeata (SE) and a fraction free of the bulk of water-soluble compounds (SF).The SE and SF samples were chemically characterized using UHPLC-ESI-QTOF-MS/MS. The anti-inflammatory activity was evaluated at three doses (SE: 30, 100, and 300 mg/kg; SF: 10, 30, and 100 mg/kg) using carrageenan-induced paw edema and pleurisy models, while genotoxicity was assessed using the SMART test.A total of 35 compounds were annotated: one cyanogenic glycoside; three glucosinolates; one benzoic acid derivative; one indole alkaloid; five spermidine alkaloids; six flavonoids; four sesquilignans; and fourteen miscellaneous compounds. Among them, seven new compounds: 4-cyano-4-hydroxybutanoic acid hexoside; tarenayanines A1, B1, C1, and D1; quercetin-sinapoyl-rhamnosyl-hexosyl-hexosyl-rhamnoside; and tarenayolone. SE and SF were not genotoxic and significantly reduced inflammatory parameters and pain sensitivity thresholds. The analgesic effect of SE was observed at 300 mg/kg, while those of SF at 100 mg/kg. SE significantly reduced edema at 100 and 300 mg/kg at 3 and/or 4 h post-induction, preventing mechanical hyperalgesia. SF inhibited edema only at 100 mg/kg, 3 and 4 h after administration. Both samples decreased leukocyte migration to pleural exudate significantly.SE and SF showed in vivo anti-inflammatory action and were found to be rich in compounds associated with this therapeutic effect, supporting the use of T. aculeata for inflammation management.