Studied the qualitative and quantitative analysis of amino acids in barley leaf powder. The results showed that there were 26 kinds of amino acids (17 kinds of hydrolyzed amino acids and 26 kinds of free amino acids) were detected in barley leaf powder. The total content of hydrolytic amino acids was 9 163.422 mg/100 g including more than 40% essential amino acids in human body. There were 9 kinds of therapeutic amino acids (Glu content was the highest, up to 1 390.873 mg/100 g) were contained in barley leaf powder. The total content of free amino acids (26 types) in barley leaf powder was 983.852 mg/100 g. The 6 kinds of free amino acids, such as Asp, Glu, Arg, Pro, His, and Val, were the main amino acids that form the flavor of barley leaf powder.