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Focusing on the antimicrobial properties of Bixa orellana Labill leaves, this research explores the essential oil extracted through hydrodistillation. Detailed chemical characterization was achieved using Gas Chromatography-Mass Spectrometry, while toxicity was assessed with Artemia salina, revealing a non-toxic classification. The study demonstrated antimicrobial efficacy against Escherichia coli and Staphylococcus aureus, with significant total phenolic content. These findings underscore the potential of this essential oil as a sustainable therapeutic alternative.