Simultaneous Purification of Pulchinenoside B4 and B5 from Pulsatilla chinensis Using Macroporous Resin and Preparative High Performance Liquid Chromatography

Pulchinenoside B4 and B5 (PB4, PB5) are two major triterpenoid saponins existing in the roots of Pulsatilla chinensis (Bunge) Regel. In this study, a systematic preparative process was developed for the simultaneous purification of PB4 and PB5 from the herb. The performance and separation characteristics of nine types of macroporous resins were critically evaluated. Static absorption/desorption experiments revealed that LX17 belonging to the polyacrylate class possessed superior separation properties. Further dynamic absorption/desorption experiments on LX17 column were conducted to obtain the optimal parameters. To obtain both compounds with high purity, a second stage procedure was developed using preparative reversed-phase high performance liquid chromatography with a dynamic axial compression column system. The separation process was high-efficiency and low-cost, which indicated potential for industrial applications.