Donkey milk is increasingly being proposed as a natural alternative milk for various categories of consumers, especially infants and the elderly population. However, its potential production, gross, and mineral composition have not been deeply investigated yet. Sixty-two individual milk samples were collected monthly from nine Ragusano donkeys reared in a specialized dairy farm. Milk yield as well as chemical and mineral composition, including macro and micro elements, were investigated over an entire lactation, from the second to the ninth month of milking. Milk yield averaged 1.64 kg/day, which highlights good aptitude of the Ragusano breed for the production of milk. Gross composition was characterized by low content of dry matter (8.19%), a high amount of lactose (6.07%), low protein (1.34%), and very low-fat content (0.16%). Whey proteins represented 58% of the total protein, and proteose-peptones accounted for 0.35 ± 0.07 g per 100 g. Total ash content was 0.36 g per 100 g and represented 4.40% of the dry matter. The most abundant element was K, which was followed by Ca, Na, and P. As expected, the micro elements Fe, Zn, and Cu were found in low amounts or in traces. Dry matter, fat, whey proteins. The total ash, Ca, P, Mg, and mineral ratios were significantly affected by the lactation stage.