We have developed and optimized a novel, efficient and accurate fingerprint method using high-performance liquid chromatography-photodiode array detection (HPLC-DAD) for the quality control of cultured Cordyceps militaris (L.) Link. The feasibility and advantages of the used chromatographic fingerprint were verified for the evaluation of cultured C. militaris by systematically comparing chromatograms with a professional analytical software recommended by State Food and Drug Administration (SFDA) of PR China. The results revealed that the chromatographic fingerprint combining similarity evaluation could efficiently identify and distinguish cultured C. militaris from different sources.