玄参中哈巴苷的制备方法研究

Preparation Methods of Harpagide from Scrophularia Ningpoensis

关键词:  玄参  哈巴苷  制备方法  大孔树脂

Key words:  Scrophularia ningpoensis  harpagide  preparation methods  macroporous resin

摘要:

目的 优选玄参中哈巴苷的制备工艺。方法 运用正交试验法，以哈巴苷提取量为指标，优化玄参中哈巴苷的提取方法；采用大孔吸附树脂法，以哈巴苷吸附-解吸率为指标，确定最佳树脂型号，优化其富集工艺；采用柱色谱法，通过对固定相、洗脱剂、上样量、色谱柱径高比、洗脱体积的考察，优化哈巴苷的纯化工艺。结果 提取工艺为玄参药材加10倍量水提取3次，每次1.5 h；富集工艺为采用SP825大孔吸附树脂柱色谱，上样液浓度为生药0.07 g·mL^-1，树脂柱径高比为1：6，吸附流速为1.0 mL·min^-1，最大比吸附量为0.40 g·mL^-1；纯化工艺为采用硅胶柱色谱法，上样量为样品：硅胶（1：70），径高比为1：15，洗脱剂为氯仿-甲醇（4：1，2：1），洗脱体积为氯仿-甲醇（4：1）洗脱2个柱体积、氯仿-甲醇（2：1）洗脱1个柱体积，再用C₁₈色谱柱（径高比1：9）纯化。结论 所建立的工艺制备效果良好，操作简单，重复性好，制备得到的哈巴苷纯度>98%，且得率较高，可用于批量生产，为玄参的进一步应用与产品开发提供参考。

Abstract:

OBJECTIVE To optimize the preparation methods of harpagide of Scrophularia ningpoensis. METHODS Orthogonal test was adopted to optimize extraction technology of harpagide with extraction capacity as the index. The macroporous resin was employed to enrich the harpagide. Taking the adsorption and desorption rates as the indexes, the best type of macroporous adsorption resin was screened and the enrichment technology was optimized. The purification technology of harpagide was optimized using column chromatography by investigating the stationary phase, eluting solvents, loading amount of sample, the diameter height ratio of chromatographic column and eluting volumns. RESULTS The optimized extraction technology of harpagide was that Scrophularia ningpoensis was extracted with water for 3 times, 1.5 h for each time. The optimized enrichment technology was that the SP825 type of macroporous adsorption resin was employed to enrich the harpagide with the following parameters:the sample concentration at 0.07 g·mL^-1, l:6 of the ratio of diameter and height, the absorptive flow rate at 1.0 mL·min^-1, and 0.40 g·mL^-1 of the maximum adsorption capacity. The optimized purification process using silica gel and C₁₈ column chrimatography was as follows:1:70 of the ratio of loading sample amount and silica gel, l:15 of the ratio of diameter and height, the eluant solvent of a mixture solvent system of chloroform and methanol, eluting with 2 BV of chloroform/methanol (4:1),then eluting with 1 BV of chloroform/methanol (2:1); the final purification using C₁₈ column chrimatography (the ratio of diameter and height was l:9). CONCLUSION The established preparation method of harpagide is convenient and good repeatability with a better effect. The purity of prepared harpagide is over 98%, and the yield is high. And the preparetion methods can be used for bulk production. It provides a reference for the further application and product development of Scrophularia ningpoensis.