Abstract:

A reversed-phase high performance liquid chromatographic (RP-HPLC) method has been presented for the simultaneous determination of 17 amino acids in the pupal exuviae of Antheraea pernyi. Norleucine was used as internal standard for quantification using the internal standard method. For amino acid analysis, a Venusil-AA amino acid column was used, and phenylisothiocyanate (PITC) was the precolumn derivatization agent. Elutions were carried out in a binary gradient mode. The detection wavelength was set at 254 nm. The calibration curves of 17 amino acids all exhibited good linearity over the range from 0.5 to 0.0125μmol. Mean recoveries for 17 amino acids were between 96.2% and 101.9%. The total amino acid content of the pupal exuviae of Antheraea pernyi was determined to be approximately 21.9%, and the total essential amino acid content represented 33.62% of the total amino acids, and the content of arginine was the highest. This method is simple and efficient and provides a good method for the determination of amino acids in he pupal exuviae of Antheraea pernyi.

Key words: high performance liquid chromatography, pre-column derivatization, phenylisothiocyanate, amino acid, norleucine, the pupal exuviae of Antheraea pernyi