RP-HPLC法测定六味五灵片中五味子醇甲、特女贞苷和连翘苷的含量

Determination of schisandrin,specnuezhenide and forsythin in Liuweiwuling Tablets by RP-HPLC

目的建立能同时检测六味五灵片中五味子醇甲、特女贞苷和连翘苷3种活性成分含量的RP-HPLC法。方法采用RP-HPLC法,色谱柱:Waters Sunfire C18(250 mm×4.6 mm,5.0μm);流速:1.0 mL·min^-1;柱温:28℃;进样体积:25μL;流动相:2 mL·L^-1磷酸(A)-乙腈(B)体系。梯度洗脱,洗脱程序为:0~7 min,89%A;7~33 min,89%A^23%A;33~35 min,23%A^89%A;35~40 min,89%A。外标法测定。结果在上述色谱条件下,五味子醇甲、特女贞苷和连翘苷均具有良好的分离度,阴性对照无干扰;3种活性成分质量浓度均在0.4~4.0μg·mL^-1范围内线性关系良好,r值均大于0.9950。平均回收率均在98.5%~99.9%范围内;重复性和精密度RSD值均小于5.0%;供试品溶液在24 h内稳定性良好。结论该方法具有前处理简单、检测速度快和准确度高等特点,适用于六味五灵片的质量控制。

Objective To develop an RP-HPLC method for simultaneous determination of the contents of 3 active ingredients(schisandrin,specnuezhenide and forsythin)in Liuweiwuling Tablets.Methods RP-HPLC method was adopted,in which Waters Sunfire C18(250 mm×4.6 mm,5.0μm)column was used for separation with column temperature set at 28℃.The flowing rate was 1.0 mL·min^-1 and injection volume was 25μL.The mobile phase was 2 mL·L^-1 phosphoric acid(A)-acetonitrile(B)system.Gradient elution program was as follows:0-7 min,89%A;7-33 min,89%A-23%A;33-35 min,23%A-89%A;35-40 min,89%A.External standard method was used for analysis.Results Schisandrin,specnuezhenide and forsythin were well separated under above chromatographic conditions,and the negative standard solution showed no interference.The 3 active components showed good linear relationships in the range 0.4-4.0μg·mL^-1 with correlation coefficients(r)all above 0.9950.The recoveries of standard addition were ranged from 98.5%to 99.9%.RSDs of repeatability and precision were less than 5.0%.Sample solution was stable within 24 h.Conclusion The method has the characteristics of simple pre-treatment,high speed and accuracy,and is suitable for the quality control of Liuweiwuling Tablets.