Abstract: Destruxins, a family of cyclic peptide mycotoxins, have been being paid more and more attention for their multiple bioactivities such as insecticide, antivirus and immunomodulation. In order to decrease the consumption of solvents and time to isolate and purify destruxins A and B (DA and DB), Ultrasonic Wave Extraction (UWV) and Vacuum Liquid Chromatography (VLC) were employed. Under the optimal conditions of pH 4-5, solvent (dichloromethane/ethyl acetate, v v 1 = 1/1)/broth (v v 1) between 1-1.5 and extracted time in 45-90 min, UWV gave an excellent recovery (extracting efficiency > 90%). Meanwhile, in the VCL experiment, the gradient of hexane/acetone 100 /0-93/7 as eluant could isolate DA and DB clearly. However, the eluants of dichloromethane/methanol and hexane/ethyl acetate were not as good as hexane/acetone.