摘要
目的:建立指纹图谱、多指标定量与多元统计分析相结合的复方银花解毒颗粒质量评价方法。方法:采用HPLC法,色谱柱为Kromasil100-5-C_(18)(250 mm×4.6 mm,5μm);流动相为乙腈-0.1%磷酸水溶液,梯度洗脱;柱温30℃;指纹图谱和含量测定检测波长分别为230、330 nm;体积流量为1.0 mL·min^(-1);进样量10μL。结合聚类分析(CA)、主成分分析(PCA)与正交偏最小二乘法-判别分析(OPLS-DA)对结果进行评价。结果:15批样品的相似度均>0.962,标定了15个共有峰,鉴定出其中9个色谱峰;CA和PCA将15批样品聚为2类;PCA显示,2个主成分的累积方差贡献率为93.6%;结合PCA和OPLS-DA,筛选出包括连翘酯苷A、绿原酸在内的5个差异标志物。8个待测成分(新绿原酸、绿原酸、隐绿原酸、连翘酯苷A、异绿原酸B、异绿原酸A、4,5-二-O-咖啡酰奎宁酸、蒙花苷)的平均加样回收率在97.6%~102.7%,RSD均<1.5%;8个成分在一定质量浓度范围内线性关系良好(r≥0.9996),结论:该方法操作简便,结果准确,重复性好,可用于复方银花解毒颗粒的质量评价。
Objective:To establish a quality evaluation method of Fufangyinhuajiedu Granules based on HPLC fingerprints,quantitative analysis of multi-components and multivariate statistical analysis.Methods:The granule components were determined by high performance liquid chromatography with a Kromasil 100-5-C_(18)(250 mm×4.6 mm,5μm)column eluted by acetonitrile(A)-0.1%phosphoric acid(B)gradiently.The flow rate was 1.0 mL·min^(-1),the injection volume was 10μL and the column temperature was 30℃.The detection wavelengths of fingerprints and content determination were set at 230 nm and 330 nm,respectively.Combined with cluster analysis(CA),principal component analysis(PCA),and orthogonal partial least squares discriminant analysis(OPLS-DA),the quality of 15 batches of Fufangyinhuajiedu Granules was analyzed.Results:The similarity of the 15 batches of Fufangyinhuajiedu Granules was greater than 0.962,and a total of 15 common peaks were calibrated and nine of them were identified.CA and PCA were used to classify the 15 batches of samples into two sub-categories.PCA showed that the cumulative variance contribution rate of the two principal components was 93.6%.Combined with PCA and OPLS-DA,five differential markers including forsythiaside A and chlorogenic acid were selected.The average recovery rates of eight components(neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,forsythoside A,isochlorogenic acid B,isochlorogenic acid A,4,5-dicaffeoylquinic acid and buddleoside)were 94.91%~108.16%,with RSD less than 1.5%.The eight components showed good linear relationships within certain concentration ranges(r≥0.9996).Conclusion:This simple,accurate and reproducible method can be used for the quality evaluation of Fufangyinhuajiedu Granules.
作者
刘光大
吴建雄
王书源
黄雁
杨琳妹
何枢衡
LIU Guangda;WU Jianxiong;WANG Shuyuan;HUANG Yan;YANG Linmei;HE Shuheng(Hefei Yifan Biopharmaceuticals Inc.,Hefei 230000,China)
出处
《药学与临床研究》
2021年第6期405-410,共6页
Pharmaceutical and Clinical Research
