Preparative Isolation of Ginseng Saponin from Panax ginseng Root Using High-speed Countercurrent Chromatography

摘要

Ginseng saponin was isolated from Panax ginseng root using high-speed countercurrent chromatography (HSCCC). Preliminary studies were performed to optimize physical properties of two-phase solvent system and operating parameters including rotation speed of column, elution mode of mobile phase, and flow rate. Two-phase solvent system for isolation of ginseng saponins was composed of chloroform, water; and methanol as blending solvent. Chloroform-aqueous methanol (4:6) systems with various concentration of methanol in water were evaluated for retention of stationary phase in column. Retention of stationary phase decreased with increasing flow rate in tail-to-head elution mode using upper phase as mobile phase and head-to-tail elution mode using lowerphase as mobile phase. Latter mode produced high retention at flow rate of 5 mL/min. Optimum conditions for isolation of saponin were chloroform/methanol/water (40/39/21) solvent system; mobile phase, of lower organic layer; flow rate, of 5 mL/min, headto tail elution mode, rotation speed, of 800 rpm, and sample injection, of 200 μL. Recovery yield of ginseng saponin from Panax ginseng root extract by HSCCC was 63.6%, and the purity of HSCCC fractions was verified by TLC.