Abstract: This study was designed to establish a method to obtain the fingerprint chromatogram for the quantitative determination of Cordyceps sinensis in different sizes, a comparison of Cordyceps sinensis from five places was made to analyze its similarity and the content of main nucleosides (uridine, inoside, guanosine and adenosine). The assay was performed on a Waters XSelect HSS T3 C₁₈ (4.6 mm×250 mm, 5 μm), with a mobile phase consisting of water (A)-acetonitrile (B) at the flow rate of 0.6 mL·min^-1 (0-5 min, 0 B; 5-15 min, 0→10% B; 15-30 min, 10%→20% B; 30-33 min, 20%→50% B; 33-35 min, 50%→0 B; 35-40 min, 0 B). The detection wavelength was 260 nm and the column temperature was set at 30℃, and the injection volume was 5 μL. The results showed that there was no significant difference of the nucleosides in samples from the same place of the different sizes, but contents of the nucleosides variate a lot by production places. More data are required for further research. The method is proved for scientific and specific formulation of the standard in evaluation of circulated Cordyceps sinensis.