基于指纹图谱及化学模式识别的归芪通脉合剂质量评价

Quality evaluation of Guiqi tongmai mixture based on fingerprint and chemical pattern recognition

目的建立归芪通脉合剂的指纹图谱,结合化学模式识别评价其质量。方法以芸香柚皮苷为参照峰,采用《中药色谱指纹图谱相似度评价系统(2012版)》建立13批归芪通脉合剂的高效液相色谱(HPLC)指纹图谱,指认共有峰并进行相似度评价;进行层次聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA),以变量重要性投影(VIP)值大于1为标准筛选影响制剂质量的差异标志物。结果13批归芪通脉合剂指纹图谱共有19个共有峰,相似度为0.928~0.990;指认了8个共有峰,分别为7号峰(羟基红花黄色素A)、8号峰(芍药内酯苷)、9号峰(芍药苷)、10号峰(毛蕊异黄酮葡萄糖苷)、12号峰(甘草苷)、14号峰(甘草酸铵)、16号峰(芸香柚皮苷)、19号峰(橙皮苷)。HCA结果显示,13批样品可分为3类,其中S5、S9、S11~S13为一类,S4、S8为一类,S1~S3、S6~S7、S10为一类。PCA结果显示,主成分1~7的累计方差贡献率为92.115%。OPLS-DA结果显示,芸香柚皮苷、芍药苷、甘草苷、甘草酸铵和11、1号峰对应成分的VIP值均大于1。结论建立了归芪通脉合剂的HPLC指纹图谱,结合化学模式识别技术可用于评价该制剂的整体质量;芸香柚皮苷等6个成分是影响归芪通脉合剂质量的差异标志物。

OBJECTIVE To establish the fingerprint of Guiqi tongmai mixture and evaluate its quality in combination with chemical pattern recognition.METHODS Using narirutin as reference peak,HPLC fingerprint of Guiqi tongmai mixture was established with Similarity Evaluation System of Chromatographic Fingerprint of TCM(2012 edition).Common peaks identification and similarity evaluation were conducted.Hierarchical cluster analysis(HCA),principle component analysis(PCA)and orthogonal partial least square-discriminate analysis(OPLS-DA)were performed,and the value of variable importance in the projection(VIP)greater than 1 was taken as the standard to screen the differential markers affecting the quality of Guiqi tongmai mixture.RESULTS Nineteen common peaks in the HPLC fingerprints of thirteen batches of Guiqi tongmai mixture were calibrated,and the similarities of them were 0.928-0.990;eight common peaks were identified,including the peak 7(hydroxysafflor yellow A),peak8(albiflorin),peak 9(paeoniflorin),peak 10(calycosin-7-O-glucoside),peak 12(liquiritin),peak 14(ammonium glycyrrhizinate),peak 16(narirutin)and peak 19(hesperidin).According to the results of HCA,thirteen batches of samples could be clustered into three categories:S5,S9 and S11-S13 were clustered into one category;S4 and S8 were clustered into one category;S1-S3,S6-S7 and S10 were clustered into one category.The results of PCA indicated that the accumulative variance contribution rate of the principle components 1-7 was 92.115%.The results of OPLS-DA showed that VIP values of narirutin,paeoniflorin,liquiritin,ammonium glycyrrhizinate,the components corresponding to peak 11 and peak 1 were greater than 1.CONCLUSIONS HPLC fingerprint of Guiqi tongmai mixture is established,and it could be used for quality evaluation of Guiqi tongmai mixture by combining with chemical pattern recognition techniques.Six components such as narirutin are differential markers affecting the quality of Guiqi tongmai mixture.