Solvent extraction method was used to extract the pleuromutilin from baked mycelium of Pleurotus mutilis which is a kind of pleuromutilin-producing strain.Based on the single screening factor test,the conditions of pleuromutilin extraction were optimized by orthogonal design.Specifically,the conditions were: solvent-biomass ratio of 4 mL/g,extraction time of 10 min,rotation speed of 140 r/min and extraction times of 3 times.94.56% of pleuromutilin extraction rate was reached under the optimum conditions.This extraction process was simple,stable and feasible,making it a practical method for industrial applications.