HPLC法同时测定腰痛片中芍药苷、川续断皂苷Ⅵ、补骨脂素和异补骨脂素含量

HPLC simultaneous determination of paeoniflorin,asperosaponin Ⅵ,psoralen and isopsoralen in Yaotong tablets

目的：建立同时测定腰痛片（杜仲叶、赤芍、补骨脂、续断、狗脊等）中活性成分芍药苷、川续断皂苷Ⅵ、补骨脂素与异补骨脂素含量的方法。方法：采用HPLC法,色谱柱为Inert Sustain C18柱（250 mm×4.6 mm,5μm）,流动相为乙腈（A）-水（B）,梯度洗脱（0 min,10%A;60 min,50%A）,流速1 m L·min-1,柱温25℃,检测波长为212 nm（川续断皂苷Ⅵ）、238 nm（芍药苷、补骨脂素和异补骨脂素）,进样量10μL。结果：芍药苷、川续断皂苷Ⅵ、补骨脂素与异补骨脂素质量浓度分别在201.6-4 032μg·m L-1（r=0.999 5）、80-2 000μg·m L-1（r=0.999 9）、11.04-220.8μg·m L-1（r=1.00 0）、8.7-174μg·m L-1（r=0.999 7）范围内与峰面积线性关系良好,平均回收率（n=9）分别为100.6%、100.6%、100.4%、99.93%,精密度试验RSD均小于0.80%。结论：经方法学验证,该法能同时测定4种成分的含量,适用于腰痛片的质量控制。

Objective：To determine the content of four major components（paeoniflorin,asperosaponin Ⅵ,psoralen and isopsoralen） in Yaotong tablets by high performance liquid chromatography.Methods：The determination was conducted on the Insert Sustain-C18column（250 mm × 4.6 mm,5 μm） with the mobile phase of acetonitrile（A）-water（B）,gradient elution（0 min,10% A;60 min,50% A） at the flow rate of 1 m L·min-1;the column temperature was at 25 ℃;the detection wavelength was 212 nm for asperosaponin Ⅵ,and 238 nm for paeoniflorin,psoralen and isopsoralen;and the injection volume was 10 μL.Results：Good linear relationship of paeoniflorin,asperosaponin Ⅵ,psoralen and isopsoralen between peak area and concentrations were noted within 201.6-4 032 μg·m L-1（r = 0.999 5）,80-2 000 μg·m L-1（r = 0.999 9）,11.04-220.8 μg·m L-1（r = 1.000）,and 8.7-174 μg·m L-1（r = 0.999 7）,respectively.The average recoveries（n = 9） were 100.6%,100.6%,100.4%,99.93%,RSD of precision test was less than 0.8%.Conclusion：Methodology verification proved that the method could be applied to simultaneously determine four components and control the quality Yaotong tablets.