4种高寒藏药材及其愈伤组织药效成分的高效液相色谱测定

J4 ›› 2010, Vol. 45 ›› Issue (1): 41-45.

4种高寒藏药材及其愈伤组织药效成分的高效液相色谱测定

赵忠娟王燕飞蔡云飞夏光敏向凤宁

赵忠娟王燕飞夏光敏向凤宁:植物细胞工程及种质创新教育部重点实验室, 山东大学生命科学学院, 山东济南 250100; 蔡云飞:山东教育学院生物科学与生物技术学院, 山东济南 250013

收稿日期:2009-07-06 出版日期:2010-01-16 发布日期:2010-03-25
通讯作者: 向凤宁(1965-),女,博士生导师,主要研究方向为细胞生物学.Email:xfn0990@sdu.edu.cn
作者简介:赵忠娟(1986-),女,硕士,主要研究方向为细胞生物学. Email:zzjfrances@yahoo.com.cn
基金资助:
国家自然科学基金资助项目(30470154,30771116);山东省杰出青年基金资助项目(GJ200810);教育部新世纪人才支持计划资助项目(NCET-05-058);教育部博士点基金资助项目(20050422015)

Determination of efficacious components of Gentianaceae plants and their calli by RP-HPLC

ZHAO Zhong-Juan, WANG Yan-Fei, JIA Guang-Min, XIANG Feng-Ning:The Key Laboratory of Plant Cell Engineering and Germplasm Innovation, Ministry of Education,Schoold of Life Science, Shandong Univeristy, Jinan 250100, Shangdong, China CAI Yun-Fei: Department of Bioscience and Biotechnology, Shandong Institute of Education,Jinan 250013, Shandong, China

Received:2009-07-06 Online:2010-01-16 Published:2010-03-25

摘要/Abstract

摘要：

采用改进的反相高效液相色谱法，快速测定了4种高寒藏药材：川西獐牙菜（Swertiamusstii Franch）、管花秦艽（Gentiana siphonantha Maxim）、白花假龙胆(Gentianellaalbiflora Maxim)、湿生扁蕾（Gentianopsis paludosa Maxim）原植株及其愈伤组织中5种有效成分：獐牙菜苦甙（Swertiamarin）、龙胆苦甙(Gentiopicroside)、芒果甙（Mangiferin）、齐墩果酸（Oleanolic acid）及熊果酸(Urolic acid)的含量。色谱柱为Phenomenex Luna C₁₈反相柱(4.6mm×250mm，5μm)；柱温：室温；獐牙菜苦甙、龙胆苦甙、芒果甙检测条件：流动相V_甲醇∶V_水∶V_冰乙酸=25.00∶75.00∶0.04，检测波长259nm，流速0.8mL/min；齐墩果酸与熊果酸检测条件：流动相V_甲醇∶V_水∶V_冰乙酸=85.00∶15.00∶0.04，检测波长209nm，流速1.0mL/min。结果表明，4种藏药材原植株药效成分种类及含量均有差别，4种藏药材中仅有川西獐牙菜含有所测定的5种药效成分单体，其余均含4种。各药效成分单体中，龙胆苦甙含量最高(1.9536%～3.7285%)。4种藏药材愈伤组织的药效成分含量均低于原植株，但同一有效成分在不同藏药材愈伤组织中含量下降的幅度不同，按生长速度推算，各药效成分的积累量均高于原植株，值得进一步开发利用。本法准确、快速，可有效的分离和检测4种高寒藏药材原植株及其愈伤组织中的药效含量，为通过细胞工程生产藏药材有效成分提供了准确、灵敏、有效的分析方法。

关键词: 反相高效液相色谱法；高寒藏药材；愈伤组织；药效成分

Abstract:

A developed reverse phase high performance liquid chromatography (RPHPLC) was used to determine the contents of five efficacious components (Swertiamarin, Gentiopicroside, Mangiferin, Oleanolic acid and Urolic acid) in Chinese medical herbs Swertia musstii Franch，　Gentiana siphonantha Maxim， and　Gentianella albiflora Maxim., Gentianopsis paludosa Ma. and their calli from subculturals. A Phenomenex Luna C₁₈ column was used and the column temperature was room temperature. Swertiamarin， Gentiopicroside and Mangiferin were detected at 259nm with Vmethanol∶Vwater∶Vacetic acid=25.00∶75.00∶0.04 as mobile phase and 0.8?mL/min as flowrate. While by Vmethanol∶Vwater∶Vacetic acid=85.00∶15.00∶0.04 as mobile phase, 1.0mL/min as flow rate and 209?nm as wavelength, and the contents of Oleanolic and Urolic acid were analyzed. The kind and content of efficacious components from the four Chinese medical herbs were quite different. Swertia -musstii-Franch contained the most efficacious components and its contents were the highest. The contents of five efficacious compounds in the above four medical herbs calli had less efficacious compounds than that of wild plants and their changes were different. However, according to the projected growth rate, the content of efficacious components accumulation of the callus was much higher than that of the wild plant. This is a precise and quick method to analyze the efficacious components inChinese medical herbs, which provides the basic for large-scale production of efficacious components in Chinese medical herbs.

Key words: reversed-phase high performance liquid chromatography (RP-HPLC); chinese medical herbs; calli; efficacious components

赵忠娟王燕飞蔡云飞夏光敏向凤宁. 4种高寒藏药材及其愈伤组织药效成分的高效液相色谱测定[J]. J4, 2010, 45(1): 41-45.

ZHAO Zhong-Juan, WANG Yan-Fei, CAI Yun-Fei, JIA Guang-Min, XIANG Feng-Ning. Determination of efficacious components of Gentianaceae plants and their calli by RP-HPLC[J]. J4, 2010, 45(1): 41-45.

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