Objective: To isolate active constituents from toad skin and investigate inhibition effect of different macroporous resin elution parts on growth of lung cancer cell line A549. Method: Type of macroporous resin was screened by static adsorption-elution test

single factor tests were adopted to optimize separation and purification technology of active constituents from toad skin.Assay methods of total alkaloids and four bufadienolides were established

inhibition rate of different elution parts on lung cancer cell line A549 was determined by MTT assay. Result: AB-8 macroporous absorption resin was selected with a better adsorption separation property for total alkaloids and four bufadienolides from toad skin

four different elution parts exhibited certain inhibition effect on lung cancer cell line A549

30% and 70% ethanol eluent showed stronger inhibition effect with IC50 of 1.83 and 2.23 mg·L-1

respectively. Conclusion: Macroporous absorption resin could be used for separation and purification of active constituents from toad skin

inhibition effect of different elution parts on lung cancer cell line A549 showed a certain concentration-dependent

30% and 70% ethanol elution parts could be used as key active ingredients in antitumor preparations of toad skin.