雷公藤药材中6种有效成分以及总二萜内酯、总生物碱、总三萜的含量测定

Determination of six effective components,total diterpenoids,total alkaloids and total triterpenes in Tripterygium wilfordii Hook.f.

目的:建立HPLC同时定量雷公藤药材中6种有效成分的方法,以及用紫外可见分光光度法测定雷公藤总二萜内酯、总生物碱、总三萜3个有效部位含量的方法,并研究不同产地的雷公藤药材中上述成分的含量差异。方法:①采用高效液相色谱法,色谱柱:Agilent ZORBAX SB-C18柱(4.6mm×250mm,5μm),以乙腈-磷酸(0.1%)-三乙胺(0.2%)为流动相进行梯度洗脱;柱温35℃;检测波长:0-65min为225nm;65min后转换为210nm;流速0.75mL/min,运行时间80min,测定11个不同产地雷公藤药材中6种有效成分的含量。②分别以雷公藤甲素、雷公藤次碱、雷公藤红素为对照,用紫外可见分光光度法测定雷公藤总二萜内酯、总生物碱及总三萜的含量。结果:不同产地的雷公藤药材有效成分及有效部位的含量差异悬殊,如雷公藤晋碱的含量相差291.6倍,总二萜内酯类高低含量之间相差34.7倍。结论:本研究所建立的方法稳定性、精密度、重复性良好,适用于雷公藤有效成分和有效部位的含量测定;不同来源的药材成分含量差异显著,对雷公藤实施多指标的含量控制是保证其临床药用安全可控的必要途径。

Objective： To determine six effective components（triptolide,triptonide,wilforgine,wilforine,tripterine,wilforlide A） by HPLC and three effective parts by UV-Vis spectrophotometry in Tripterygium wilfordii Hook.f.,then study their contents in raw materials from different areas,which offers a reliable method to scientifically evaluate and effectively control the quality of Tripterygium wilfordii Hook.f.and its preparation.Methods： The HPLC analysis was carried out on a Agilent ZORBAX SB-C18 column（4.6mm×250mm,5μm） with the mobile phase consisting of acetonitrile-water（0.1% phosphoric acid and 0.2% triethylamine） in gradient mode.The column temperature was kept at 35℃ and the detect wavelength was set at 225nm from 0 minute to 65 minute and then changed to 210nm after 65 minute;The flow rate was 0.75mL/min and the analytic time was 80min,which was used to determine the six effective components in samples produced by ten different areas.Using triptolide,wilforine and tripterine as the contrast,total diterpenoids,total alkaloids and total triterpenes was determined by UV-Vis spectrophotometry.Results： There were significant differences of effective components and effective parts in Tripterygium wilfordii Hook.f.from different areas.For example,the highest content of wilforgine was about 291.6 times of the lowest one,while the highest content of the total diterpenoids was about 34.7 times of the lowest one.Conclusion： The method is in good reproducibility,precision and stability and is fit to determine the six effective ingredients in Tripterygium wilfordii Hook.f.At the same time,significant content difference has also been shown among different areas.It is very important to control the quality with multi-index for clinic safety.