Separation and Purification of Harpagoside and Sibirioside A from Extract of Scrophulariae Radix by Macroporous Resin and High-speed Counter-current Chromatography

OBJECTIVE To study the purification method of harpagoside and sibirioside A using macroporous resins combined with high-speed countercurrent chromatography (HSCCC) from crude extract of Scrophulariae Radix. METHODS The crude extract of Scrophulariae Radix separated through macroporous resin in order to enrich the target compounds preliminary. Then the target compounds separated by high-speed countercurrent chromatography (HSCCC) with a two-phase solvent system contained n-butanol-ethyl acetate-water (1∶9∶10). The upper phase was stationary phase while the lower phase was mobile phase with a flow rate of 1.5 mL·min^-1 and the wavelength detection was set at 210 nm. RESULTS Harpagoside and sibirioside A were obtained from the crude extract of Scrophulariae Radix, and contents were 98.1% and 97.2%, respectively. The chemical structures of harpagoside and sibirioside A were identified by ¹H-NMR and ¹³C-NMR. CONCLUSION The results reveal that the method is high preparative capacity and high efficiently in separation of harpagoside and sibirioside A from Scrophulariae Radix.