The invention relates to a liquid chromatography method for separating and determining an N-succinyl tryptophan enantiomer. The current detection method is low in efficiency and poor in accuracy. Theliquid chromatography method adopts a protein bonding type chiral HPLC column which mainly takes ammonium acetate-acetonitrile as a flow phase to perform separation and determination for the N-succinyl-tryptophan enantiomer. The efficient liquid phase separate detection method of the N-succinyl tryptophan enantiomer can effectively separate and determine the N-succinyl tryptophan enantiomer, is high in specificity and high in accuracy, and is suitable for qualification and quantification of the N-succinyl tryptophan enantiomer.