经典名方桂枝加葛根汤标准汤剂HPLC指纹图谱研究

HPLC fingerprint of classic prescription Guizhi Jia Gegen Decoction

目的建立经典名方桂枝加葛根汤物质基准的HPLC指纹图谱,结合化学模式识别方法对其质量进行分析与评价,为其质量控制提供参考。方法采用HPLC建立桂枝加葛根汤指纹图谱,采用中药色谱指纹图谱相似度评价软件(2012年版),并结合聚类分析(clustering analysis,HCA)、主成分分析(principal component analysis,PCA)与正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)对结果进行分析,评价不同批次桂枝加葛根汤物质基准的质量差异,并找寻造成批间质量差异的主要化学成分。结果建立了桂枝加葛根汤物质基准的HPLC指纹图谱,确定了29个共有峰,并对各共有峰进行药材归属,其中3~14、16~19、21~25号峰来自葛根;26号峰来自桂枝;1、15号峰来自白芍,20、27~29号峰来自甘草,2号峰来自大枣。通过与对照品比对指认了其中11个成分,分别为1号峰没食子酸、8号峰3′-羟基葛根素、11号峰葛根素、12号峰葛根素芹菜糖苷、13号峰3′-甲氧基葛根素、15号峰芍药苷、16号峰大豆苷、20号峰甘草苷、22号峰染料木苷、26号峰肉桂酸、28号峰甘草酸铵。15批桂枝加葛根汤物质基准的HPLC指纹图谱与对照指纹图谱的相似度均>0.94;3种化学模式识别方法均可将样品分为2类;筛选出了11种主要差异成分,其中已指认出的为葛根素、3′-甲氧基葛根素、葛根素芹菜糖苷、大豆苷、3′-羟基葛根素、没食子酸。结论建立的指纹图谱分析方法灵敏度高、稳定性强、数据准确可靠,可为经典名方桂枝加葛根汤物质基准的质量评价提供依据。

Objective To establish a HPLC fingerprint of the substance benchmark of classic prescription Guizhi Jia Gegen Decoction(GJGD,桂枝加葛根汤)and evaluate its quality with chemical pattern recognition method,so as to provide reference for the quality control of GJGD.Methods HPLC was used to establish the fingerprint of GJGD,and traditional Chinese medicine(TCM)chromatographic fingerprint similarity evaluation software(2012)combined with clustering analysis(HCA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to evaluate the quality differences between different batches of substance benchmark of GJGD,and find the main chemical composition causing quality differences between batches.Results The HPLC fingerprint of GJGD was established,29 common peaks were identified,and the medicinal materials of each common peak were assigned,peaks 3—14,16—19,21—25 came from Gegen(Puerariae Lobatae Radix);peak 26 came from Guizhi(Cinnamomi Ramulus);peaks 1 and 15 came from Baishao(Paeoniae Radix Alba);peaks 20,27—29 came from Gancao(Glycyrrhizae Radix et Rhizoma),and peak 2 came from Dazao(Jujubae Fructus).Eleven of them were identified by comparing with the control substance,which were gallic acid(peak 1),3′-hydroxy puerarin(peak 8),puerarin(peak 11),puerarin apioside(peak 12),3′-methoxypuerarin(peak 13),paeoniflorin(peak 15),daidzin(peak 16),liquiritin(peak 20),genistin(peak 22),cinnamic acid(peak 26),monoammonium glycyrrhizinate(peak 28).The similarity between the HPLC fingerprints of 15 batches of GJGD and the control fingerprints was>0.94.The three chemical pattern recognition methods could divide the samples into two categories.Eleven compounds were screened out,of which puerarin,3′-methoxypuerarin,mirificin,daidzin,3′-hydroxypuerarin and gallic acid were identified among them.Conclusion The established fingerprint analysis method has high sensitivity,strong stability,accurate and reliable data,which can provide a basis for the quality evaluation of GJGD.