Abstract: Objective:HPLC fingerprints of Ophiocordyceps sinensis from different areas were established to provide a scientific basis for quality control of O. sinensis and cultivated mycelium. Method:Chromatographic fingerprints of 10 batches of O. sinensis and 9 batches of cultivated mycelium were established by HPLC with 0.3% acetic acid and methanol as mobile phase for gradient elution;the detection wavelength was set at 260 nm. The fingerprints were analyzed by using similarity evaluation,principal component analysis(PCA)and cluster analysis(CA). Results:The reference HPLC fingerprint of O. sinensis was established with 18 common peaks,and 5 of common peaks were identified,including cytidine,uridine,inosine,guanosine and adenosine. The peak area RSD of precision,stability and repeatability of the method were less than 2.99%,3.04% and 3.16%,respectively,which conformed to the requirements of fingerprint test. The similarity of 10 batches O. sinensis was greater than 0.92.There were some differences between O. sinensis and cultivated mycelium,with the similarity value from 0.563 to 0.693.The results of PCA and CA were consistent with similarity analysis,which all samples were classified into 3 groups and 10 batches of O. sinensis belonged to 1 group. Conclusion:The HPLC fingerprint combined with PCA and CA methods could be quickly used to identify genuine O. sinensis,and provide scientific basis to evaluate the quality of O. sinensis.