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.