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

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(Swertia musstii Franch)、管花秦艽(Gentiana siphonantha Maxim)、白花假龙胆(Gentianella albiflora 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(4.6 mm×250 mm,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259 nm,流速0.8 mL/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209 nm,流速1.0 mL/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。

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

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, Centiana siphonantha Maxim, and Centianlla albiflora Maxim., Gentianopsis paludosa Ma. And their calli from subculturals. A Phenomenex luna C_18 column was used and the column temperature was room temperature. Swertiamarin, Gentiopicroside and Mangiferin were detected at 259nm with V_(methanol):V_(water):V_(acetic acid) = 25.00:75.00:0.04 as mobile phase and 0.8 mL/min as flow rate. While by V_(methanol):V_(water):V_(acetic acid)=85.00:15.00:0.04 as mobile phase, 1.0 mL/min as flow rate and 209 nm as wavelength, and the contents of Oleanolie and Urolic acid were analyzed. The kind and content of effica cious components from the four Chirlese 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 berbs 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 pre-cise and quick method to analyze the efficacious components in Chinese medical herbs,which provides the basic for large-scale production of efficacious components in Chinese medical herbs.