SEPARATION AND PURIFICATION OF TRITERPENE SAPONINS FROM ROOTS OF RADIX PHYTOLACCAE BY HIGH-SPEED COUNTERCURRENT CHROMATOGRAPHY COUPLED WITH EVAPORATIVE LIGHT SCATTERING DETECTION.

Coupled with evaporative light scattering detection, high-speed countercurrent chromatography was successfully applied for the first time for separation and purification of four triterpene saponins including esculentosides A, B, C, and D from the roots of Radix phytolaccae. The separation was performed with an optimized two-phase solvent system composed of chloroform-methanol-water (4:4:2,v/v), using the lower phase as the mobile phase at a flow rate of 1.5 mL/min. From 150 mg of crude extract, 46.3 mg of esculentoside A, 21.8 mg of esculentoside B, 7.3 mg of esculentoside C, and 13.6 mg of esculentoside D were obtained at purities of 96.7%, 99.2%, 96.5%, and 97.8%, respectively, as determined by HPLC analysis. The structures of the four triterpene saponins were identified by ESI-MS,1H NMR and 13C NMR. [ABSTRACT FROM AUTHOR]