基于UPLC指纹图谱和多指标成分定量的不同产地白茅根标准汤剂质量评价研究

Study on quality evaluation of Imperatae Rhizoma standard decoction based on UPLC fingerprint and multi-components content determination

本研究通过超高效液相色谱(UPLC)指纹图谱结合多成分定量对不同产地的白茅根标准汤剂的质量进行评价。采用相似度计算、聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)对指纹图谱数据进行分析。结果显示,18批白茅根标准汤剂指纹图谱共标识出9个共有指纹峰,指纹图谱相似度在0.920~0.997;HCA将18批样品分为3类,河北产区的样品自成一类,江苏与安徽、河南的样品在分类上存在交叉;PCA结果显示,除江苏外,同一产区白茅根标准汤剂指纹图谱共有峰的差异较小;OPLS-DA筛选出3个差异标志性色谱峰,分别为峰9(对羟基肉桂酸)、峰6和峰5(绿原酸)。对白茅根标准汤剂中绿原酸、新绿原酸、隐绿原酸和对羟基肉桂酸的含量进行测定,结果显示,4种酚酸的总含量:河北>安徽>江苏>河南,不同产区的样品4种酚酸的总含量差异明显;其中河南产区的样品绿原酸、新绿原酸和隐绿原酸的含量明显低于其他产区,安徽产区的样品对羟基肉桂酸的平均含量最高且比较稳定,江苏产区的样品对羟基肉桂酸和4种酚酸的总含量波动较大。所建立的分析方法稳定可靠,重复性好,为白茅根标准汤剂及其相关制剂的质量评价提供参考。

In this study, the quality of Imperatae Rhizoma standard decoction from different habitats was evaluated by UPLC fingerprint and quantitative analysis of multiple components.Similarity calculation, cluster analysis(HCA),principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used to analyze the fingerprint data.The results showed that there were 9 common fingerprint peaks in the fingerprints of 18 batches of Imperatae Rhizoma standard decoction.The similarity of fingerprints ranged from 0.920 to 0.997.HCA divided 18 batches of samples into three categories.The samples from Hebei Province were of their own kind.The samples from Jiangsu Province were crossed with those from Anhui and Henan provinces respectively.PCA showed that, except Jiangsu Province, the differences of common peaks in the fingerprints of Imperatae Rhizoma standard decoction in the same production area were small.OPLS-DA screened three differential marker chromatographic peaks, which were peak 9(p-hydroxycinnamic acid),peak 6 and peak 5(chlorogenic acid) respectively.The contents of chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid and p-hydroxycinnamic acid in Imperatae Rhizoma standard decoction were determined.The results showed that the total content of four phenolic acids in Imperatae Rhizoma standard decoction were: Hebei > Anhui > Jiangsu > Henan.The contents of four phenolic acids in samples from different production areas were significantly different.The contents of chlorogenic acid, neochlorogenic acid and cryptochlorogenic acid in the samples from Henan production area were significantly lower than that from other production areas.The average content of p-hydroxycinnamic acid in the samples from Anhui production area was the highest and relatively stable.The total amount of p-hydroxycinnamic acid and four phenolic acids in the samples of Jiangsu production area fluctuated greatly.The established analytical method is stable, reliable and reproducible, which can provide reference for the quality evaluation of Imperatae Rhizoma standard decoction and its related preparations.