Virola surinamensis is an abundant floodplain tree, popularly known as ucuuba, that grows in the Amazon. In this study, ucuuba seed oil was obtained by supercritical fluid extraction under different operating conditions, as well as Soxhlet extraction. The operating conditions for supercritical extraction were an extraction temperature of 40, 60, or 80 degrees C, a pressure of 350bar, and a CO2 mass flow of 7.9x10(-5) kg/s. The supercritical extraction curves were fitted to mass transfer models, and the fatty acid profiles of the extracts were determined by gas chromatography. The antimicrobial activity was assessed against Candida albicans, Staphylococcus aureus, and Escherichia coli. The highest yield obtained using supercritical CO2 was 64.39% and the lowest was 59.21%. The phytochemical analysis showed the presence of steroids, terpenes, coumarins, and phenolic compounds. All ucuuba oil samples showed antioxidant activity. Regarding the antimicrobial activity, ucuuba oil only showed activity against S. aureus.