Butter is a widely consumed dairy product, and its nutritional composition can vary significantly depending on the production method. This study aimed to analyze and compare the fatty acid composition of 25 traditional and 25 industrial butter samples sourced from various regions across Algeria. The analysis focused on saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), degree of unsaturation, and peroxidability index. Fatty acid content was determined using capillary gas chromatography following the American Oil Chemistry Society (AOCS) standard procedures. The results revealed significant differences between traditional and industrial butter. Traditional butter had a lower SFA content (48.93 ± 0.2%) compared to industrial butter (61.14 ± 0.2%). Conversely, traditional butter showed a higher MUFA content (31.01 ± 0.2%) compared to industrial butter (28.65 ± 0.2%). Similarly, traditional butter had a markedly higher PUFA content (10.03 ± 0.1%) than industrial butter (6.67 ± 0.1%). Moreover, the degree of unsaturation and the peroxidability index were also greater in traditional butter (15.82 and 19.34, respectively) compared to industrial butter (12.85 and 15.52, respectively). These findings suggest that traditional butter offers a more favorable fatty acid profile, with higher unsaturated fat content, which may have implications for consumer health and product quality. Further studies should investigate the potential health benefits of traditional butter and explore the impact of local production methods on its nutritional properties.