The chemical composition and physicochemical properties of hexane and ethyl acetate extracts of skin, juice, and seeds of Opuntia dillenii fruit collected from three Moroccan regions (Oujda, Nador, and Essaouira) were studied. The study revealed that the seed oil extracts presented the highest yield of 13.12%, followed by the skin fraction (1.77%) and the juice extract (0.49%). The evaluation of fatty acid compositions using GC-MS analysis revealed the presence of linoleic acid as a dominating unsaturated fatty acid with a value of 72.39%, followed by palmitic acid, oleic acid, and stearic acid in all localities. Otherwise, the juice extract of Oujda locality was richer in margaric acid (37.41%), followed by Essaouira skin extract (10.7%) and Oujda seed extract (6.18%). However, the campesterol was detected only in trace in the juice extract. The physicochemical properties of O. dillenii seed oils such as acid value, peroxide value, ester value, pH value, saponification value, density, and refractive index were all found to be in good agreement with the quality criteria for pure and fresh oils. In addition, principal component analysis (PCA) and hierarchical cluster analysis (HCA) were implemented to compare the difference in the chemical composition of the different O. dillenii extracts.