木芙蓉叶中总黄酮类成分的纯化工艺研究  

Purification process of total flavonoids from Hibiscus mutabilis leaves

作　　者：邓琴 陈晓兰 万静 徐剑 吴静澜 谢树才 胡娟 黄耀磊 谌文元 DENG Qin;CHEN Xiaolan;WAN Jing;XU Jian;WU Jinglan;XIE Shucai;HU Juan;HUANG Yaolei;CHEN Wenyuan(School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;Bijie Medical College,Bijie 551700,China)

机构地区：[1]贵州中医药大学药学院,贵州贵阳550025 [2]毕节医学高等专科学校,贵州毕节551700

出　　处：《贵州科学》2025年第2期46-54,共9页Guizhou Science

基　　金：国家苗药工程技术研究中心能力提升(黔科合中引地[2023]006);贵州省高层次创新型人才项目(黔科合平台人才-GCC[2023]037);贵州省普通高等学校科技拔尖人才支持计划项目(黔教合KY字[2018]051)。

摘　　要：目的:优选木芙蓉叶总黄酮的聚酰胺-大孔树脂联用纯化工艺,并建立木芙蓉叶中黄酮类组分的HPLC分析方法,为该有效部位的开发提供参考。方法:以木芙蓉叶中总黄酮纯度为指标,采用单因素联合正交试验优化木芙蓉叶总黄酮的纯化工艺,考察的工艺参数包括上样液浓度、最大上样量、上样流速、水洗用量、洗脱剂浓度及用量、洗脱流速、聚酰胺与大孔树脂柱体积比等,确定最佳纯化工艺。结果:木芙蓉叶总黄酮的最佳纯化工艺为:聚酰胺树脂10 g,浓度为0.16 g生药/mL的木芙蓉叶提取液100 mL,上样流速为1.0 mL/min,先用10 BV的蒸馏水洗,然后用2.5 BV 20%的乙醇除杂,将除杂后的聚酰胺加入AB-8大孔树脂顶部,聚酰胺与大孔树脂柱体积比为1∶2,用4 BV(聚酰胺体积)80%的乙醇以1.5 mL/min的流速进行洗脱,收集洗脱液,水浴挥干,得到木芙蓉叶总黄酮纯化物,纯化后,木芙蓉叶总黄酮纯度达70%左右。木芙蓉叶中5种黄酮类单体成分(芦丁、金丝桃苷、异槲皮苷、槲皮素、山柰酚)的HPLC分析方法为:乙腈-0.5%冰乙酸,梯度洗脱;检测波长:370 nm;柱温:35℃;流速:1 mL/min;进样量:10μL。结论:聚酰胺-大孔树脂联用对木芙蓉叶中总黄酮的纯化效果好,操作简单,效率高,稳定性好,适用于木芙蓉叶中总黄酮提取物的纯化。In this study we optimized the purification process of total flavonoids from the leaves of Hibiscus mutabilis by polyamide-macroporous resin,and established an HPLC method for the analysis of flavonoids in the leaves of Hibiscus mutabilis.The purification process of total flavonoids from Hibiscus mutabilis leaves was optimized by single factor and orthogonal tests with the purity of total flavonoids as the index,and the process parameters investigated included the concentration of the injection solution,the maximal injection volume,the injection flow rate,the concentration and amount of the eluent,the flow rate of the elution,and the ratio of the column volume of polyamide to macroporous resin,etc.The optimal purification process was determined as follows:10 g polyamide resin,100 mL of Hibiscus mutabilis leaves extract at a concentration of 0.16 g/mL,injection flow rate of 1.0 mL/min,distilled water wash of 10 BV,impurity removal by 2.5 BV of 20%ethanol,the volume ratio of the polyamide to the macroporous resin was 1:2,and the elution was carried out with 4 BV(volume of polyamide)of 80%ethanol at a flow rate of 1.5 mL/min.Under the above conditions,the purity of the total flavonoids from Hibiscus mutabilis leaves was about 70%after purification.The HPLC analysis parameters of the five flavonoid monomers(rutin,chrysin,isoquercitrin,quercetin,and kaempferol)in the leaves of Hibiscus mutabilis was as follows:acetonitrile-0.5%glacial acetic acid,gradient elution,detection wavelength of 370 nm,column temperature at 35℃,flow rate at 1 mL/min,and injection volume of 10μL.The method has the advantages of simple operation,high efficiency and good stability,and is suitable for the purification of total flavonoids from Hibiscus mutabilis leaves.

关 键 词：木芙蓉叶 总黄酮 纯化 大孔树脂 聚酰胺 

分 类 号：R284.2[医药卫生—中药学]