ABSTRACT: Sunflower seeds (Helianthus annuus L.) are rich in lipids, particularly fatty acids such as linoleic acid, and in vitamins, notably tocopherol (vitamin E), a potent antioxidant. This study aimed to evaluate the oil yield of sunflower seeds cultivated in non-traditional soils and to characterize their physical, chemical, and microbiological properties. Oil extraction was performed using a Soxhlet apparatus with diethyl ether and petroleum ether as solvents. Microbiological analyses were conducted using two reference microorganisms, Candida albicans and Escherichia coli, following the methodologies outlined in the Clinical and Laboratory Standards Institute (CLSI) guideline M7-A6. The average oil yield from dehulled seeds was approximately 50%, with accession 2286 exhibiting the highest lipid content (34.68%). To assess oil quality, several parameters were measured, including acidity index, peroxide value, pH, density, free fatty acid content, ash content, moisture content, volatile matter, and infrared (IR) spectral characteristics. Statistical significance of the results was determined using analysis of variance (ANOVA). The antimicrobial assay showed no inhibitory activity of sunflower oil against either E. coli or C. albicans, indicating a lack of antimicrobial properties under the tested conditions.