In this study,a high performance liquid chromatography (HPLC) method for the simultaneous determination of xanthine,guanine,adenine and hypoxanthine in Stropharia rugosoannulata was disclosed.Firstly,the content of one purine standard is detected,and then the relative correction factor and relative retention time between the purine standard and the other three purine components in the test sample of S. rugosoannulata were calculated.Through formula calculation,it was convenient to realize one measurement and multiple evaluation of four purine components in of S. rugosoannulata.The method has the advantages of simple operation,high sensitivity,low standard cost and simplified experimental steps.At present,there is no content determination method for S. rugosoannulata and its related products.The research results have certain reference value for the identification and quality control of S. rugosoannulata.