HSCCC分离纯化未成熟罗汉果皂苷类化合物

Separation and purification of mogrosides from unripe Siraitia grosvenorii by high-speed countercurrent chromatography

关键词:  高速逆流色谱, 未成熟罗汉果, 罗汉果皂苷ⅡE, 11-O-罗汉果皂苷Ⅱ, 罗汉果皂苷Ⅲ, 11-O-罗汉果皂苷Ⅲ

Key words:  high-speed countercurrent chromatography(HSCCC), unripe Siraitia grosvenorii, mogroside Ⅱ E, 11-O-mogroside Ⅱ, mogroside Ⅲ, 11-O-mogroside Ⅲ

摘要:

为建立高速逆流色谱分离未成熟罗汉果中皂苷类化合物的方法,该研究将罗汉果粗提物先经过大孔树脂富集皂苷类化合物,再采用高速逆流色谱分离罗汉果皂苷。结果表明:以氯仿-甲醇-正丁醇-水(5:6:1:4,v/v/v/v)作为两相溶剂系统,上相为固定相,下相为流动相,在主机转速为860 r·min^-1,流速为2.5 mL·min^-1,检测波长为203 nm的条件下,一次性制备得到4个化合物,即11-O-罗汉果皂苷Ⅱ(Ⅰ)、罗汉果皂苷ⅡE(Ⅱ)、11-O-罗汉果皂苷Ⅲ(Ⅲ)和罗汉果皂苷Ⅲ(Ⅳ),经高效液相色谱检测纯度分别为95.5%、98.2%、80.1%和97.6%。该方法实现了未成熟罗汉果皂苷快速有效的分离,具有样品回收率高、损失少、避免样品失活等优点,提高了分离效率。该研究结果为更多的罗汉果皂苷化合物的分离纯化奠定了基础,补充与优化了罗汉果皂苷类化合物的分离方法。

Abstract:

We established a method for separating mogrosides from unripe Siraitia grosvenorii by High-speed countercurrent chromatography(HSCCC). Crude mogrosides fraction was enriched by macroporous resin method,and then separated by HSCCC. The two-phase solvent system composed of chloroform-methanol-n-butanol-water(5:6:1:4, v/v/v/v)was used for separation. The upper phase was used as the stationary phase, while the lower phase as the mobile phase. The rotation speed was 860 r·min^-1, the flow rate was 2.5 mL·min^-1, and the detection wavelength was 203 nm. 11-O-mogroside Ⅱ, mogroside Ⅱ E, 11-O-mogroside Ⅲ and mogroside Ⅲ, four compounds were prepared in one step,with the purity of 95.5%,98.2%,80.1% and 97.6%, respectively. This method that has the advantages of high sample recovery rate, low loss and avoiding sample inactivation, which successfully realized the rapid and effective separation of mogrosides. Hence, HSCCC can improve separation efficiency and provide theoretical foundation for the separation and purification of moremogrosides, complementing and optimizing the method for separating mogrosides.