Donkey milk (DM) has been considered a valuable alternative to human and bovine coun-terparts as well as to infant formulas. Milk extracellular vesicles (EVs) have been proposed to influence key biological processes. The purpose of this study is to provide a compre-hensive characterization of the protein composition of extracellular vesicles (EVs) by ex-tending quantitative proteomic comparisons to EVs derived from donkey colostrum (DC) and mature donkey milk (MDM). The EVs were isolated from DC and MDM samples, characterized, and subjected to proteomic analysis using the tandem mass tag-based quantitative approach