This study investigates the influence of breed on the physico-chemical and mineral properties of milk from Simmental and Holstein cows in the Dukagjin plain region of Kosovo. Milk samples were collected from five municipalities within the region, and their composition was analyzed using standardized laboratory methods. The results demonstrate a significant breed effect on milk fat content, with Simmental cows producing milk with higher fat percentages compared to Holsteins. In contrast, protein content did not show a significant difference between the two breeds, while milk from Holstein cows exhibited higher acidity levels. Additionally, environmental factors such as topography and the lactation stage were found to impact milk composition, with higher values observed in areas with extensive pastureland. No significant breed-related differences were detected in mineral content, although mineral levels varied according to local relief conditions. These findings contribute to the understanding of breed-specific milk characteristics in the Dukagjin region and provide valuable insights for dairy farmers aiming to optimize milk production.