Abstract: To compare the protein contents, as well as the amino acids and their contents in the Rana dybowskii jelly coat from Jingyu, Dunhua, Jiaohe, Fusong, Huadian and at different ages. Protein content was determined by Coomassie method. And the type, and the method for the determination of 21 kinds of hydrolyzed amino acids in Rana dybowskii jelly coat by high performance liquid chromatography with pre-column derivatization of Phenylisothiocyanate as derivatization reagent was established.The results showed that the pre-column derivatization reversed-phase high performance liquid chromatography with Phenylisothiocyanate as a derivatizing reagent was highly efficient, stable and reproducible, and it was suitable for the determination of amino acids in Rana dybowskii jelly coat.The contents of protein and amino acids in Rana dybowskii jelly coat from three-year-old were higher than those from two-year-old. There were significant differences in protein and amino acid contents among different origins.The highest protein, amino acid contents and the ratio of essential amino acids were found in Jiaohe three-year-old. The protein content was ( 174.33 ± 4.55) mg/g; the total amount of amino acids was ( 10.004 ±0.20) mg/g; and the essential amino acids accounted for 24.57%.The lowest protein, amino acid contents were found in Husong two-year-old, the contents were ( 147.39 ± 1.09) mg/g, ( 6.414 ± 0.10) mg/g.The lowest ratio of essential amino acids produced in Huadian at the age of 2 forest frog film in 20.54%.The content of protein, the total amount of amino acids and the proportion of essential amino acids were 26.94 mg/g, 3.59 mg/g and 4.03% respectively.