益胃汤物质基准的HPLC指纹图谱分析及多组分含量测定

HPLC Fingerprint Analysis and Multi-component Determination of Substance Benchmark of Yiweitang

目的:建立益胃汤物质基准的高效液相色谱法(HPLC)指纹图谱和多组分含量测定方法,并结合化学模式识别方法对其质量进行评价。方法:制备15批益胃汤物质基准,采用"中药色谱指纹图谱相似度评价系统"(2012版)计算相似度,利用聚类分析、主成分分析和正交偏最小二乘法-判别分析对15批样品指纹图谱的共有峰进行处理,评价不同批次样品间的质量差异;对梓醇、毛蕊花糖苷和麦冬甲基黄烷酮A的含量进行测定,流动相系统为乙腈-磷酸水溶液,检测波长设定了210 nm和334 nm。结果:益胃汤物质基准的HPLC指纹图谱有22个共有峰,其中1,9,12,14~17,19和20号峰归属生地黄,3,4,6,7和21号峰归属于北沙参,5和22号峰归属于麦冬,2和18号峰归属于玉竹,8号峰为麦冬和生地黄共有峰,10号峰为麦冬,玉竹和生地黄共有峰,11号峰为4味药的共有峰,13号峰为玉竹和生地黄共有峰。15批益胃汤物质基准的HPLC指纹图谱与对照指纹图谱的相似度均>0.90;3种化学模式识别方法均可将15批样品分为四类。定量分析结果表明15批益胃汤物质基准中梓醇、毛蕊花糖苷及麦冬甲基黄烷酮A的质量分数范围分别为0.37%~1.14%,0.002%~0.054%,0.016%~0.079%。结论:建立的益胃汤物质基准指纹图谱和含量测定方法分离度好、准确度高,体现了益胃汤物质基准整体化学成分特征,可为该经典名方复方制剂的研发及质量控制提供实验依据。

Objective: To establish high performance liquid chromatography(HPLC)fingerprint and multi-component determination for the substance benchmark of Yiweitang, and to evaluate its quality in combination with chemical pattern recognition method. Method: Fifteen batches of substance benchmark of Yiweitang were prepared,the "Chinese medicine chromatographic fingerprint similarity evaluation system"(2012 edition)was used to calculate similarity. Cluster analysis,principal component analysis and orthogonal partial least squares-discriminant analysis were employed to handle the common peaks for evaluating the quality difference among 15 batches of the substance benchmark. The contents of catalpol, verbascoside and methylophiopogonanone A were determined with mobile phase system of acetonitrile-phosphoric acid solution at detection wavelengths of 210 nm and 334 nm. Result:There were 22 common peaks in HPLC fingerprint of the substance benchmark,among them,peaks 1,9,12,14-17,19 and 20 belonged to Rehmanniae Radix,peaks3,4,6,7 and 21 belonged to Glehniae Radix,peaks 5 and 22 belonged to Ophiopogonis Radix,peaks 2 and 18 belonged to Polygonati Odorati Rhiaoma,peak 8 was the common peak of Ophiopogonis Radix and Rehmanniae Radix,peak 10 was shared by Ophiopogonis Radix,Polygonati Odorati Rhiaoma and Rehmanniae Radix,peak 11 was the common peak of these four herbs,and peak 13 was shared by Polygonati Odorati Rhiaoma and Rehmanniae Radix. The similarities between HPLC fingerprints of 15 batches of the substance benchmark and the control fingerprint were all >0.90,the samples could be divided into four categories by three chemical pattern recognition methods. Quantitative analysis showed that the contents of catalpol, verbascoside and methylophiopogonanone A among 15 batches of samples ranged from 0.37% to 1.14%,0.002% to 0.054% and 0.016% to 0.079%,respectively. Conclusion: The established fingerprint and determination for the substance benchmark of Yiweitang have good separation and high accuracy,which reflect the overall chemical composition characteristics of Yiweitang,and can provide experimental basis for the further development and quality control of the compound preparations of this famous classical formula.