Objective: To establish a quality analysis method for Venenum Bufonis and to study influence of different producing area on active composition. Method: TLC and HPLC were applied with analysis and determination of the content of cinobufagin and resibufogenin. Result: Venenum Bufonis from different producing area had the similar compositions. But the content of compositions was significantly different especially for resibufogenin in that the Venenum Bufonis produced in Shandong had the highest content of cinobufagin and resibufogenin. The difference of compositions displayed majorly in spot 2(resibufogenin)

spot 3(cinobufagin)

spot 4

spot 6 and spot 9 in TLC chromatogram and peak 3

peak 4

peak 6

peak 7

peak 8

peak 9

peak 14 and peak 15 in HPLC chromatogram. The HPLC chromatogram could also see that the content of peak 7

peak 8 and the content of peak 14(cinobufagin)

peak 15(resibufogenin)had possible relationship. Conclusion: The quality of Venenum Bufonis from different producing area has significant difference. The established method is stable

feasible with a good reproducibility and specificity

and suitable for identification and quantitative analysis of Venenum Bufonis.