Objective To detect quality the nucleosides contained in cultured Cordyceps militaris mycelia from different habitats and natural Cordyceps sinensis for qulity evaluation. Methods The contents of nucleosides in 11 batches of cultured Cordyceps militaris from 9 different places and 4 types of natural Cordyceps sinensis were determined by using a HPLC method with the evaluating markers of uridnine,adenosine and cordycepin. Results The contents of nucleosides in cultured Cordyceps militaris mycelia were varied from 0.27% to 0.87% and those in Cordyceps sinensis were 0.17% to 0.25%.Cordycepin levels differed more obviously,less or no cordycepin was detected in natural Cordyceps sinensis. Conclusion Different contents of nucleosides are presented in cultured Cordyceps militaris mycelia from different habitats,while much higher than those in natural Cordyceps sinensis.Among which,the contents of adenosine and uridnine are constant,and could be used as valuable bases for development and quality evaluation of artificial Cordyceps militaris mycelia.