基于HPLC指纹图谱及多成分定量对刺五加的质量评价

Quality evaluation of Acanthopanax senticosus based on HPLC fingerprint and multi-component content determination

目的建立刺五加Acanthopanax senticosus HPLC指纹图谱及原儿茶酸、松柏苷、紫丁香苷、绿原酸、刺五加苷B1、刺五加苷E、丁香树脂酚7种成分含测定方法。方法采用Diamonsil 5μm C18(2)色谱柱(4.6 mm×250 mm,5μm),乙腈-0.1%磷酸水溶液为流动相,梯度洗脱;检测波长210 nm,柱温30℃,体积流量0.8 mL/min,进样量10μL。通过“中国色谱指纹图谱相似度评价系统”(2012版),建立了刺五加HPLC指纹图谱,同时测定了7种成分的含量。结合主成分分析、正交偏最小二乘法判别分析和皮尔逊相关性分析对刺五加进行了质量评价。结果建立了刺五加HPLC指纹图谱,共确定了18个共有色谱峰;原儿茶酸、松柏苷、紫丁香苷、绿原酸、刺五加苷B1、刺五加苷E以及丁香树脂酚线性关系良好,r2均大于等于0.9991,平均回收率分别为92.51%、97.63%、95.11%、101.08%、105.29%、98.92%、85.44%、94.89%。主成分分析提取了2个主成分,其方差累积为66.920%,表明这2个主成分可包含原有数据的大部分信息;正交偏最小二乘法判别分析筛选出刺五加样品中2个差异性成分(紫丁香苷、刺五加苷B1);皮尔逊相关性分析显示紫丁香苷、绿原酸和刺五加苷B1与其他化学成分的相关性最强。结论建立的刺五加HPLC指纹图谱及多成分含量测定方法简单、准确、重复性好,可用于刺五加的质量评价,为其质量控制提供依据。

Objective To establish the HPLC fingerprint of Acanthopanax senticosus and determination method of seven major constituents such as protocatechuic acid,coniferin,syringin,chlorogenic acid,eleutheroside B1,eleutheroside E,and syringaresinol.Methods The chromatography was performed on the Diamonsil 5μm C18(2)chromatographic column(4.6 mm×250 mm,5μm)with acetonitrile-0.1%phosphoric acid aqueous solution as the mobile phase and gradient elution;the detection wavelength was 210 nm,the column temperature was 30°C,the flow rate was 0.8 mL/min,and the injection volume was 10μL.The“Chinese Chromatographic Fingerprint Similarity Evaluation System”(2012 Edition)was used to establish the HPLC fingerprint of A.senticosus,and the contents of the seven components were simultaneously determine.A combination of principal component analysis(PCA),orthogonal partial least squares discriminant analysis(OPLS-DA)and Pearson’s correlation analysis was used to evaluate the quality of A.senticosus.Results HPLC fingerprint of A.senticosus was established,and a taotal of 18 common chromatographic peaks were identified.Protocatechuic acid,coniferin,syringin,chlorogenic acid,eleutheroside B1,eleutheroside E,and syringaresinol showed good linearity with r2 greater than or equal to 0.9991,and the average recoveries of these were 92.51%,97.63%,95.11%,101.08%,105.29%,98.92%,and 94.89%,respectively.Two principal components were extracted by the PCA,and the cumulative percentage of variance was 66.920%,which indicated that these two principal components could contain most of the information of the original data;the OPLSDA screened out the two differential components(syringin and eleutheroside B1)in the samples of A.senticosus.Pearson’s correlation analysis showed that syringin,chlorogenic acid and eleutheroside B1 had the strongest correlation with other chemical components.Conclusion The established HPLC fingerprint and multi-component content determination method of A.senticosus are simple,accurate and reproducible,which can be used for the quality evaluation of A.senticosus and provide the basis for its quality control.