The aims of this study were to perform a detailed analysis of the chemical composition by gas chromatography, cinematic viscosity in function of temperature by viscometer Schott AVS 350, and biological tests of abóbora (Cucurbita argyrosperma) seed oil, collected in Mucajai city, Roraima state, Brazil. Thus, the abóbora seed samples were manually submitted to the oven at 50 °C for 72 hours, and crushed and sieved between 20-40 Mesh. The seed oil extraction was carried out in a Soxhlet apparatus with hexane for 3 hours, providing an oil yield of 29.8%. In the results were identified a total of 8 fatty acids by gas chromatography; of these 33.8% are saturated fatty acids and 65.7% are unsaturated fatty acids: palmitic (21.5%), stearic (11.6%), oleic (37.0%), linoleic (28.1%) and linolenic (0.3%). In the biological test, the oil was more active against Staphylococcus aureus (29.01%), Listeria monocytogenes (34.56%), Pseudomonas aeruginosa (23.86%) and Citrobacter freundii (32.74%), which features a characteristic low therapeutic index for these bacteria. The kinematic viscosity of the oil reduces in function of temperature (298.15 to 353.15 K) in an exponential curve, demonstrating to refer to a Newtonian fluid.