Abstract: OBJECTIVE To establish a method for the determination of 17 amino acids contents in different grades of antler pieces from Cervus nippon using pre-column derivatization-HPLC, and to compare the difference of amino acid contents. METHODS The contents of 17 kinds of amino acids in different grades of antler pieces were detected by HPLC with pre-column derivatization and gradient elution with phenyl isothiocyanate. RESULTS There were significant differences in the total amino acid contents of different grades of antler pieces. Amino acid contents from high to low was wax, powder, sand and bone chips. CONCLUSION The quality of antler pieces is affected by the position of harvest. The method is suitable for the determination of the amino acid contents in the antler pieces, which can provide a theoretical basis for the formulation of antler pieces.