Objective: To investigate the effects of HgCl2 pollution on the physiological status and bufadienolides level of toad by using liquid chromatography-mass spectrometry (LC-MS/MS)

and provide references for quality evaluation of Chinese medicine venenum bufonis. Method: The different concentrations of HgCl2 solution (0.5

0.75 and 1.0 g·L-1) were prepared to treat the toad with semi-static water method

and then their physiological status was observed. Venenum bufonis was collected after 30 days. Furthermore

ultra performance liquid chromatography-mass spectrometry was used to determine the level of bufadienolides in toad venom and HemI heat map software was used for data analysis. Result: The results showed that HgCl2 solution could significantly influence the bufadienolides level. As compared with normal group

20 kinds of bufadienolides were up-regulated in low concentration (0.5 g·L-1) Hg2+ exposure group

and there were 12 kinds of bufadienolides with 1.5 fold change. However

15 kinds of bufadienolides in 0.75 g·L-1 Hg2+ exposure group and 13 kinds in 1.0 g·L-1 Hg2+ exposure group were down-regulated

and most of them were medium and low fat-soluble toad steroids (bufarenogin

dehydrated cinobufagin

etc.). Conclusion: These illustrated that Hg2+ exposure could affect the levels of bufadienolides in toad venom

showing an obvious decrease tendency with the increase of HgCl2 solution concentration.