Objective To improve the extraction and isolation technology for neogambogic acid from Gamboge. Methods The extracting process was optimized by orthogonal design. Alcohol was used as extraction solvent,AB8 macroporous resin was employed to enrich extracts, silica gel G was used as stationary phase in wet chromatography column with a mobile phase consisted of petroleum etheralcohol (20∶1).Finally，a golden yellow crystal compound was obtained. The structure of the obtained compound was identified by melting point (mp) determination, ultraviolet spectra (UV), infrared spectra (IR), mass spectra (mass spectra,MS), 1Hnuclear magnetic resonance (NMR),and 13CNMR. Results The data of mp,UV,IR,MS, 1HNMR, and 13CNMR for the obtained golden yellow crystal compound were in accordance with those recorded in related literatures, correspondingly. Conclusion The improved technology is feasible for the extraction and isolation of neogambogic acid from Gamboge,and is beneficial to industrialization production.