The invention discloses a high performance liquid chromatography method and application of sodium 3-(benzothiazol-2-ylthio)-propanesulfonate. The method comprises the following steps: (a) providing a 3-(benzothiazol-2-ylthio)-propanesulfonate (ZPS) reference substance solution and a test sample solution; (b) analyzing the ZPS content of the test sample solution in step (a) with HPLC-UV according to an external standard method. Detection conditions are as follows: a chromatographic column uses octadecyl silane bonded silica gel as a filler; a mobile phase is formed by an organic phase and a phosphate buffer, the pH of a phosphate buffer is equal to 5.5 plus/minus 0.1, and the volume ratio of the organic phase to the phosphate buffer is (10 to 90) to (80 to 20); the detection wavelength is between 190nm and 330nm. The technical problems of imperfect measurement method of ZPS content, poor selectivity of iodometric method and poor accuracy are alleviated. The method can accurately detect the ZPS content, so that the precision is good, and the accuracy is high.