一测多评法同时测定清热颗粒中5种成分的含量

Simultaneous Determination of Five Components in Qingre Grabules(清热颗粒)by Quantitative Analysis of Multi-Components by Single Marker

目的:建立一测多评法同时测定清热颗粒中绿原酸、隐绿原酸、秦皮乙素、木犀草苷、蒙花苷5种指标性成分的含量。方法:采用HPLC法,色谱柱为Welchrom C_(18)(150 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,流速为1.0 mL/min,柱温为30℃,检测波长为320 nm,进样量为10μL。以绿原酸为内参物,分别采用多点校正法、斜率法以及定量因子3种校正方法建立各成分与绿原酸的相对校正因子,计算各成分的量,并与外标法比较,验证一测多评方法的准确性和可行性。结果:在一定线性范围内,多点校正法计算的隐绿原酸、秦皮乙素、木犀草苷、蒙花苷的f_(k/s)分别为1.0507、1.2231、1.5087、1.4374;斜率法计算的f_(k/s)分别为1.0469、1.2191、1.5051、1.4394;定量因子分别为27555502.04、26332161.83、22602724.56、18307697.59、19143636.44。比较3种一测多评法与外标法测定的成分含量差异,一测多评法与外标法检测具有良好一致性。结论:本试验建立了清热颗粒中5种成分的一测多评法,经方法学验证,一测多评法可用于清热颗粒的质量控制。

Objective:To establish quantitative analysis of multi-components by single marker(QAMS)method for simultaneous determination of five components(chlorogenic acid,cryptochlorogenic acid,aesculetin,luteoloside and linarin)in Qingre grabules.Methods:HPLC method was adopted.The determination was performed on Welchrom C_(18) column(150 mm×4.6 mm,5μm)with gradient elution by acetonitrile-0.1%phosphoric acid solution,at a flow rate of 1 mL/min.The column temperature was 30℃and the detection wavelength was 320 nm.The sample size was 10μL.With chlorogenic acid as the internal reference,three calibration methods,multi-point calibration method,slope method and quantitative factor,were used to establish the relative calibration factor of each component with chlorogenic acid.The contents of five components in Qingre grabules were determined by both QAMS and external standard method.QAMS was used to calculate the content of each component and compare it with the external standard method to verify the accuracy and feasibility of the QAMS method.Results:The RCFs of albiflorin,by multi-point calibration method,f_(k/s) of cryptochlorogenic acid,aesculetin,luteoloside and linarin were 1.0507,1.2231,1.5087 and 1.4374,respectively;by slope correction method,fk/s were 1.0469,1.2191,1.5051 and 1.4394,respectively;by quantitative factors method,quantitative factors were 27555502.04,26332161.83,22602724.56,18307697.59 and 19143636.44,respectively.The content differences was compared among the three components determined by QAMS methods and the external standard method.The results calculated with QAMS were consistent with the results by the external standard method.Conclusion:The method of QAMS for five components in Qingre grabules was established.The methodology validation showed that the method can be used for quality control of Qingre grabules.