Milk composition is an imperative aspect influencing the quality of dairy products. The study was aimed to compare the cheddar cheese made from buffalo and cow milk with respect to composition and proteolysis (water-soluble nitrogen, non-protein-nitrogen) during ripening. The assessment of proteolysis in cheese through RP-HPLC was also core part of present exploration. Considerable differences were found in compositional profile of both cheeses. Buffalo milk cheese was relatively higher in fat (30.32%) and protein (31.91%) contents throughout storage period. The rate of proteolysis and peptide's production was also comparatively higher in buffalo milk cheese as evident from water-soluble nitrogen (16.86%) and non-protein-nitrogen (8.44%) contents. Significant (p square 0.01) increase in proteolysis index was observed over the ripening period of 180 days. RP-HPLC studies demonstrated that cheeses expressed peptide peak developments from the very first day till the end of ripening. However, slight differences were observed concerning the number, area and height of peaks in cheese extracts during ripening. Hence, it was concluded that buffalo milk is superior with regard to compositional profile and proteolytic activities.