Cantharidin,a well-known natural toxin and vesicant,is extracted from the blister beetle of Mylabris phalerata Pallas or Mylabris cichorii Linnaeus.Cantharidin is the active terpenoid compound of blister beetle and displays many kinds of helpful bioactivities in vivo or in vitro.An efficient method was built successfully for the rapid extraction,separation and purification of cantharidin from the blister beetle by ultrahigh pressure-assisted extraction (UPE) in combination with high-speed counter-current chromatography (HSCCC).The UPE conditions including,extraction time,extraction pressure,solvent ratio were optimized with an orthogonal test.The crude acetone extraction was separated by silica gel column with pure chloroform as elution solution.The separated fraction containing cantharidin was further purified by HSCCC with a two-phase solvent system composed of petroleum ether–acetate–methanol–water (1.2:0.8:0.9:1.1,v/v).15.2 mg of cantharidin was obtained from 200 mg of separated fraction with the purity of 99.1% as determined by HPLC.The chemical structure of cantharidin was identified by ESI-MS,1H-NMR and 13C-NMR.