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Stryphnodendron adstringens, called ‘barbatimão’, is used in Brazilian folk medicine given its healing, astringent, anti-inflammatory, and antimicrobial properties. However, data on its safe use remain inconclusive due to controversy about the risk posed by its ethnopharmacological intake to human health. Thus, the present study aims to quantify the main classes of compounds found in S. adstringens bark powder and investigate the biological activity of ‘barbatimão’ bark decoction against pathogenic bacterial strains. We also investigated its toxic activity using the Artemia salina test and its cytogenotoxicity using the Allium cepa test. Phenols (24.5 ± 1.5 mg mL-1), tannins (15.08 ± 0.02 mg mL-1), and flavonoids (1.33 ± 0.02 mg mL-1) were quantified in the S. adstringens decoction. This decoction exhibited antibacterial activity against Gram-positive strains and toxicity against A. salina. It was not possible to observe the S. adstringens bark decoction genotoxicity in the A. cepa roots because the cell cycle stopped at the prophase, indicating the decoction’s anti-proliferation effect. S. adstringens is a plant with great herbal potential; therefore, other study models must be used to investigate its potential antibacterial and anti-proliferation applications