An efficient separation method of scopolamine from Hindu Datura extracts was successfully developed by combining liquid-liquid extraction, macroporous resins, and crystallization. First, the extraction solution was successively performed with liquid-liquid extraction by acid water and carbon tetrachloride. Secondly, D151 resin was selected from six tested resins, and the separation parameters were optimized. Finally, scopolamine hydrobromide was obtained after crystallization at -20 degrees C overnight. Through one run of above combined treatments, the content of scopolamine increased from 3.02% (w/w) to 99.12% (w/w). In addition, the products were assessed by high-performance liquid chromatography (HPLC).