Due to the increasing consumption of ghee in the Western countries, a complete characterization of buffalo and cow ghee was performed to complement and update the available literature. Ghee is a lipophilic dairy product with 98.9% lipids, 0.3% water and less than 0.9% nonfat solids. Fatty acids are the major lipid fraction and represent 85.1% and 83.65% for buffalo and cow ghee, respectively. More than 52% of the fatty acids were saturated, and palmitic (24-28.8%), stearic (9.4-14%) and myristic (8.5-10%) acids were predominant. Monounsaturated fatty acids were approximately 23.8% and the major component was oleic acid. Polyunsaturated fatty acid content was 2.45% (buffalo) and 4% (cow). The vaccenic acid (2.18%) and the conjugated linoleic acid (CLA cis-9, trans-11) with a concentration of 0.77% in buffalo and 1% in cow ghee, were the main ruminant trans fatty acids. The physicochemical and microbiological characteristics of cow and buffalo ghee complied with the literature and national regulation. Finally, the sensory profile of buffalo and cow ghee was defined with a predominantly lactic odor, followed by cooked and fatty notes. The taste was characterized as fatty, lactic, sweet and cooked; and the texture was described as fatty with fatty mouthfeel, lumpy and greasy notes.