藤络宁胶囊指纹图谱的建立及化学模式识别研究

Establishment of HPLC Fingerprints and Study on Chemical Pattern Recognition for Tengluoning Capsules

目的:建立藤络宁胶囊HPLC指纹图谱,并结合化学模式识别方法评价藤络宁胶囊的质量。方法:采用Agilent 5 TC-C_(18)色谱柱(250 mm×4.6 mm,5μm)进行色谱分离,流动相为甲醇-0.1%甲酸水溶液梯度洗脱,流速为1.0 ml·min^(-1),柱温30℃,检测波长345 nm,进样量为10μl。采用《中药色谱指纹图谱相似度评价系统(2012A版)》对指纹图谱进行相似度评价,确定共有峰,结合聚类热图分析、主成分分析和正交偏最小二乘-判别分析等模式识别方法,对不同批次藤络宁胶囊进行质量评价。结果:10批藤络宁胶囊HPLC指纹图谱有18个共有峰,相似度均在0.950以上;通过聚类分析将样品按不同批次分为2大类;主成分分析与聚类分析结果一致;最后采用正交偏最小二乘-判别分析筛选出了不同批次藤络宁胶囊的5个差异标志物。通过与对照品图谱比对,确定了3号峰为绿原酸、7号峰为东莨菪内酯、13号峰为丹酚酸B。结论:建立的指纹图谱方法稳定、可靠、重复性好,可用于藤络宁胶囊的质量控制。

Objective:To establish the HPLC fingerprints of Tengluoning capsules and evaluate its quality with chemical pattern recognition method.Methods:An Agilent 5 TC-C_(18)column(250 mm×4.6 mm,5μm)was used with methanol-0.1%formic acid aqueous solution as the mobile phase at a flow rate of 1.0 ml·min^(-1).The column temperature was 30℃,the detection wavelength was set at 345 nm,and the injection volume was 10μl.The similarity evaluation of fingerprints was performed by Similarity Evaluation System of TCM Chromatographic Fingerprint(2012 A edition)to confirm common peaks.Quality evaluation of Tengluoning capsules with different batches was performed by cluster heat map analysis(CA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA).Results:Ten batches of Tengluoning capsules had 18 common peaks in the HPLC fingerprints,and the similarities were all above 0.950.The samples were divided into two categories according to different batches through CA,and the results of PCA and CA were consistent.Five different markers of Tengluoning capsules were screened by OPLS-DA.By comparing with the reference compounds,peak 3 was identified as chlorogenic acid,peak 7 was scopoletin,and peak 13 was salvianolic acid B.Conclusion:The established fingerprint method is stable,reliable and reproducible,and can be used for the quality evaluation of Tengluoning capsules.