This study investigated the changes in chemical composition, nitrogen fraction distribution, and AA profile of milk samples obtained during lactation from the Jiangyue breed of donkey in Northwest China. Results showed that donkey milk contained 9.53% total solids, 1.57% protein, 1.16% fat, 6.33% lactose, and 0.4% ash on average, which is more similar to mare and human milk than to the milk of other mammals. Throughout the lactation investigated, pH and density were constant, protein and ash content showed an apparent negative trend (an increase in lactose content during 120 d postpartum, followed by a decrease), fat content exhibited wide variability, and variations in the content and percentage of whey protein, casein, and AA were small. The casein to whey protein ratio of 52:37 was between the lower value of human milk and the higher value of cow milk. Sodium dodecyl sulfate-PAGE results demonstrated that donkey milk is rich in beta-lactoglobulin and lysozyme. The percentages of 8 essential AA in protein of donkey milk were 38.2%, higher than those of mare and cow milk; donkey milk also had higher levels of serine (6.2%), glutamic acid (22.8%), arginine (4.6%), and valine (6.5%) and a lower level of cystine (0.4%).