Objective: To establish a HPLC method for simultaneously determining the contents of three nucleosides

namely uridine

guanosine and adenosine

and compare the content differences of the three nucleosides and the composition ratios in Corydalis Rhizoma from different regions and commercial grades. Method: A Venusil MP C18 (2) chromatographic column (4.6 mm×250 mm

5 μm) was used

with methanol (A)-water (B) as the mobile phase for gradient elution (0-10 min

1%-5%A; 10-15 min

5%-15%A; 15-22 min

15%-17%A; 22-26 min

17%-20%A; 26-32 min

20%-24%A)

at a flow rate of 1.0 mL·min-1. UV detection wavelength was set at 260 nm

and the temperature of the column was maintained at 35℃. The above HPLC method was applied to determine 38 batches of Corydalis Rhizoma and a cluster analysis was carried out for the results. Result: The contents of three nucleosides in Corydalis Rhizoma were determined

and showed a good linear relationship within the linear range (r > 0.999 8). The average recovery rate was in the range of 99.63%-104.24%

with the RSD raging from 0.7%to 2.3%. The three nucleosides could be detected in all 38 batches of Corydalis Rhizoma. There were significant differences in contents of uridine

guanosine and adenosine and the composition ratios for Corydalis Rhizoma from different regions; there were significant differences in contents of the three nucleosides and composition ratios for Corydalis Rhizoma from different farmer households of a same region. 38 batches of Corydalis Rhizoma were divided into three categories by cluster analysis. Conclusion: Overall

the contents of three nucleosides in Corydalis Rhizoma showed significant difference while there was no significant difference in nucleosides content for the Corydalis Rhizoma with different commercial grades.