不同产地车前草HPLC指纹图谱及化学模式识别研究

HPLC Fingerprint Analysis and Chemistry Pattern Recognition of Plantaginis Herba from Different Regions

目的建立车前草的高效液相色谱(HPLC)指纹图谱。方法采用HPLC法。色谱柱为Agilent 5 TC-C_(18),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,检测波长为330 nm,流速为1.0 ml/min,柱温为25℃,进样量为10μl。以9号峰为参照,绘制16批车前草药材样品HPLC指纹图谱;采用《中药色谱指纹图谱相似度评价系统》(2012版)进行相似度评价,确定共有峰;运用主成分分析(PCA)和偏最小二乘法-判别分析(PLSDA)对其进行模式识别研究。结果 16批车前草药材样品共有19个共有峰,相似度在0.854~0.957之间,聚类分析和主成分分析结果均显示,16批车前草药材样品可聚为2类,S6~S9、S13、S14聚为一类,S1~S5、S10~S12、S15、S16聚为一类。PLS-DA分析结果显示,大车前苷、毛蕊花糖苷、芹菜素、木犀草素等5个成分VIP值>1,可能是引起16批车前草药材样品质量差异的主要成分。结论本文结合化学计量学和HPLC指纹图谱,为建立全面有效的车前草药材质量控制模式提供参考。

Objective To establish a HPLC fingerprint of Plantaginis herba.Methods HPLC with gradient elution was adopted.The determination was performed on Agilent 5 TC-C_(18) column with mobile phase consisted of 0.1%acetic acid-acetonitrile.The detection wavelength was 330 nm;the flow rate was 1.0 ml/min;the column temperature was 25℃,and sample size was 10μl.HPLC fingerprints of 16 batches of samples were determined by using No.9 peak as reference.The similarity evaluation was carried out by using the TCM Chromatographic Fingerprint Similarity Evaluation System(2012 edition)so as to determine the common peaks.The differences among the samples were identified by chemical pattern recognition methods including principal component analysis(PCA)and partial least squares-discriminate analysis(PLS-DA).Results There were 19 common peaks in HPLC fingerprints of 16 batches of Plantaginis herba.The similarity of 16 batches of Plantaginis herba was between 0.854 and 0.957.16 batches of Plantaginis herba could be divided into 2 categories by cluster analysis and PCA,among which S6-S9,S13 and S14 were grouped into one category,and the remaining batches were grouped into another category.5 components with variable importance(VIP)values>1 were screened by PLS-DA,which may be the main components that caused the quality difference of 16 batches of Plantaginis herba samples.Conclusion The combination of chemometrics(CMM)and HPLC fingerprints can provide a reference for the comprehensive and effective quality control of Plantaginis herba.