Abstract:

In order to efficiently separate high-purity active components from Andrographis paniculata, a high-speed counter-current chromatography （HSCCC） procedure was developed for the preparative separation and purification of four bioactive compounds from the crude extract of the plant in a simultaneous process. The procedure was performed in a two-phase solvent system of n-hexane-ethyl acetate-ethanol-water （ in a volume ratio of 1 ： 4：2.3 ： 2.7 ） at a flowrate of 2.0 mL/min. By determining the physicochemical properties and according to the results of MS, ¹ HNMR and ¹³ CNMR, the four separated compounds were identified as 14-deoxy-andrographolide-19-β-D-glucosides （ Ⅰ ）, neoandrographolide （ Ⅱ ）, andrographolide （ Ⅲ ） and 14-deoxy-andrographolide （ Ⅳ ）. Moreover, 56. 8, 1.5, 108.3 and 218.0 mg of compounds Ⅰ - Ⅳ were respectively prepared from 500mg of crude extract by the proposed procedure, with the purities of 98.5% , 92.5% , 96.2% and 93.6% and the recoveries of 95.5% , 97.8%, 93.2% and 91.9%, respectively.

Key words: Andrographis paniculata, high-speed counter-current chromatography, active component, separation, purification, liquid chromatography-mass spectrum, nuclear magnetic resonance