Abstract: The content of α-lactoalbumin,β-lactoglobulin and lactoferrin in different kinds of milk was determined and compared by HPLC-MS in this study. The results showed that there were some differences in α-lactoalbumin,β-lactoglobulin,lactoferrin and total protein of different sources of milk. The average value of whey protein in goat milk was 0.385 g/100 g,which was lower in cow milk and yak milk. The highest content of lactoferrin was in goat milk,with an average of 4.43 mg/100 g and a maximum of 9.90 mg/100 g,which was a high-quality source of lactoferrin. The total protein content of yak milk(3.61%)was significantly higher than that of cow milk(3.22%)and goat milk(2.89%);The content of whey protein in milk was the highest among the three types of samples.