大孔吸附树脂-高速逆流色谱法分离纯化玄参中哈巴俄苷和斩龙剑苷A

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

关键词:  大孔吸附树脂  高速逆流色谱  玄参  哈巴俄苷  斩龙剑苷A

Key words:  macroporous resins  high-speed countercurrent chromatograghy(HSCCC)  Scrophulariae Radix  harpagoside  sibirioside A

摘要:

目的 以玄参的干燥根为原料，建立大孔吸附树脂-高速逆流色谱法(HSCCC)分离纯化玄参中哈巴俄苷与斩龙剑苷A的方法。方法 玄参粗提物先经过大孔吸附树脂初步分离，富集目标化合物。然后以正丁醇-乙酸乙酯-水(1∶9∶10)为溶剂体系，上相为固定相，下相为流动相，流速1.5 mL·min^-1，检测波长210 nm，利用制备型HSCCC分离纯化哈巴俄苷和斩龙剑苷A。结果 经过大孔吸附树脂-高速逆流色谱法分离后，一次性得到哈巴俄苷和斩龙剑苷A，经HPLC检测其纯度分别为98.1%和97.2%。经¹H-NMR和¹³C-NMR鉴定结构为哈巴俄苷和斩龙剑苷A。结论 该方法能够简便、高效地制备玄参中的哈巴俄苷和斩龙剑苷A。

Abstract:

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.