HPLC-MS/MS检测杜仲中绿原酸等4种活性成分的分析方法

杜仲作为一种传统的中药材,其活性成分主要包括绿原酸、桃叶珊瑚苷、京尼平苷和京尼平苷酸等。本研究建立了一种HPLC-MS/MS(高效液相色谱质谱联用仪)检测的方法,可同时检测杜仲中桃叶珊瑚苷、京尼平苷、京尼平苷酸和绿原酸的含量。建立的色谱条件:色谱柱XD-C18,流动相为0.1%甲酸水溶液和甲醇,流速0.15mL/min,柱温30℃。质谱条件: ESI离子源作为裂解源,裂解温度280℃,源内电压4kV,鞘气辅助气为氮气,流速30psi,雾化气为氦气,流速10psi。结果表明:4种目标化合物的色谱峰分离情况良好,质谱鉴定为目标物;绿原酸、桃叶珊瑚苷、京尼平苷和京尼平苷酸标准曲线的相关性系数分别为0.9935、0.9942、0.9969、0.9948;最低检测限的范围是7.603~8.853pmol;加样回收率范围是91.838%~108.326%。测定得到杜仲叶中桃叶珊瑚苷的含量为(20.552±4.032)ng/mg干质量,京尼平苷酸含量为(6.913±0.654)ng/mg干质量,绿原酸含量为(25.986±3.412)ng/mg干质量,京尼平苷含量为(0.205±0.015 )ng/mg干质量。利用该方法在7种植物组织中进行方法验证,测定得到4种目标化合物的含量,表明本方法有效且适用性广。 

A method of HPLC-MS/MS to determine chlorogenic acid and other three kinds of active components in Eucommia ulmoids

Eucommia ulmoids is a kind of traditional Chinese herbal medicine. Aucubin (AU), chlorogenic acid (CA), geniposidic acid (GA) and geniposide (G) are important components in E. ulmoids. A HPLC-MS/MS method was established to determine these four target chemicals in E. ulmoids. Chromatographic conditions were: the chromatographic column XD-C18 was adopted, the mobile phase was 0. 1% formic acid aqueous solution and methanol, the flow rate was 0. 15 mL/min, and the column temperature was 30℃. Mass spectrometry conditions were: ESI ion as a pyrolysis source, the pyrolysis temperature was 280℃, the source voltage was 4 kV, sheath gas was nitrogen, flow rate was 30 psi, the atomization gas was helium, and the flow rate was 10 psi. The R2 of CA, AU, G and GA standard curve were 0. 993 5, 0. 994 2, 0. 996 9 and 0. 994 8, respectively. Lower limit of detection was from 7. 603 to 8. 853 pmol. Spiked recovery was from 91. 838% to 108. 326%. In E. ulmoids, the content of AU was 20. 552 ± 4. 032 ng/mg dry weight ( DW) , the content of GA was 6. 913 ± 0. 654 ng/mg DW, the content of CA was 25. 986 ± 3. 412 ng/mg DW, and the content of G was 0. 205 ± 0. 015 ng/mg DW. This method was also used to determine the contents of these four target chemicals in other seven plant species, and the results indicated that the method is effective and widely applicable.