Objective To compare the HPLC measurement method for analysis amifostine, which validated by China Ministry of Public Health and United States Pharmacopeia (USP 35). Methods The HPLC methods for determination of amifostine, validated by USP 35 and China Ministry of Public Health Department were comparatively described. Amifostine and its related substances were determined with the mobile phase of 7:18 (methanol: 0.94g/L hexanesulfonic acid sodium solution), with flow rate of 1.0ml/min and 1:1 (methanol: 3.5 mmol/L octanesulfonic acid sodium solution, with flow rate of 0.7ml/min, respectively. Wave length of the two standards' detection was 220 nm. Results Both of the two standards were meet analysis requirement of amifostine and its related substances. Conclusion USP35 is better for the quality control of amifostine.