HPLC法同时测定少林正骨精中5种成分的含量及近红外定量模型的建立

Simultaneous Determination of Five Components in Shaolin Zhenggu Essence by HPLC and Establishment the NIR Quantitative Model

目的:建立HPLC法同时测定少林正骨精中阿魏酸、羌活醇、蛇床子素、异欧前胡素、二氢欧山芹醇当归酸酯含量的方法;应用近红外光谱技术(NIRS)建立少林正骨精定量分析模型。方法:采用中普红PX-C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相乙腈(A)-0.1%磷酸(B),梯度洗脱,柱温30℃,流速1.0 mL/min,检测波长320 nm;通过近红外光谱技术,采集样品的光谱图,运用OPUS6.5光谱工作站结合含量测定数据建立少林正骨精定量分析模型。结果:阿魏酸在2.22~222.06μg/mL、羌活醇2.32~231.66μg/mL、蛇床子素5.92~592.02μg/mL、异欧前胡素2.17~217.45μg/mL、二氢欧山芹醇当归酸酯2.73~273.16μg/mL范围内线性关系良好,平均加样回收率分别为97.35%、100.16%、101.65%、101.39%、96.36%,RSD分别为1.62%、1.99%、1.23%、1.79%、1.97%;少林正骨精定量分析模型初步建成,5个成分的RMSECV分别为:0.345、0.842、1.060、0.735、0.887,RMSEP分别为:0.554、1.230、2.040、0.922、0.725,外部验证预测值与化学测定值的RSD<13.10%。结论:该HPLC法经方法学验证,可用于少林正骨精中阿魏酸、羌活醇、蛇床子素、异欧前胡素、二氢欧山芹醇当归酸酯的含量测定;该近红外定量分析模型可用于产品均匀性考察。

Objective:To establish an HPLC method for simultaneous determination of ferulic acid,notoptero l.osthole,isoimperatorin.columbianadin in Shaolin Zhenggu Essence;To establish a quantitative analysis model by NIRS.Methods:The Zhongpuhong C_(18)Column(250mm×4.6 mm,5μm)was used with the mobile phase of acetonitrile(A)-0.1%phosphoric acid solution(B)in a gradient mode at 30℃,The flow rate was 1.0 mL·min^(-1)and the detection wavelength was set at 320nm;Spectrograms were collected by NIRS and a quantitative analysis model was established by OPUS 6.5 spectral workstation combined with content determination data.Results:The linear ranges of ferulic acid,notopterol,osthole,isoimperatorin,columbianadin were 2.22~222.06μg/mL,2.32~231.66μg/mL,5.92~592.02μg/mL,2.17~217.45μg/mL,2.73~273.16μg/mL.The average recoveries(n=9)were 97.35%,100.16%,101.65%,101.39%,96.36%and the RSD were 1.62%,1.99%,1.23%,1.79%,1.97%respectively.The quantitative analysis model of Shaolinzhenggujing has been preliminarily established,5 components RMSECV were 0.345,0.842,1.060,0.735,0.887 respectively,RMSEP were 0.554,1.230,2.040,0.922,0.725 respectively,the predicted values and the chemical measured values were less than 13.10%.Conclusion:The HPLC proved by method validation,the method could be used to determine contents of ferulic acid、notopterol、osthole、isoimperatorin、columbianadin in Shaolinzhenggujing.The NIR quantitative analysis model can be used to investigate products uniformity.