欧矢车菊根HPLC指纹图谱的建立及化学模式识别分析

Establishment of high-performance liquid phase(HPLC)fingerprint of Centaurea behen.L root and analysis of chemical pattern recognition

目的建立欧矢车菊根的高效液相(High performance liquid phase,HPLC)指纹图谱,结合化学模式识别法进行分析,建立多指标成分含量的测定方法。方法采用WondaSil C 18色谱柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.1%甲酸水(B)为流动相梯度洗脱,流速1 mL/min,检测波长为290 nm,柱温为30℃,进样量为10μL。利用《中药色谱指纹图谱相似度评价系统(2012版)》建立15批欧矢车菊根药材的HPLC指纹图谱并分析相似度,通过对照品比对指认,确定化学成分并进行含量测定,使用SPSS 26.0和SIMCA-P 14.1软件进行聚类分析(Hierarchical calcluster analysis,HCA)、主成分分析(Principal component analysis,PCA)分析和正交偏最小二乘判别分析(Orthogonal partial least squares discriminant analysis,OPLS-DA)分析,对不同批次样品进行质量评价。结果建立了15批欧矢车菊根HPLC指纹图谱共匹配出20个共有峰,通过对照品比对指认出绿原酸(11号峰)、咖啡酸(13号峰)、异绿原酸A(19号峰)。建立了绿原酸和异绿原酸A含量测定方法;指纹图谱相似度范围为0.601~0.983;HCA将15批欧矢车菊根分为2类;PCA得到4个主成分的累积方差贡献率为85.260%;OPLS-DA表明绿原酸和异绿原酸A是影响欧矢车菊根药材质量的差异性标志物。结论指纹图谱结合化学模式识别技术可快速筛选出欧矢车菊根药材的差异性特征成分,为欧矢车菊根药材的质量控制及标准制定提供了理论依据。

Objective:Toestablish a high-performance liquid phase(HPLC)fingerprint of Centaurea behen L.root medicinal material,toevaluate it using chemical pattern recognition method,and toestablish a multi index component content determination method.Methods WondaSil C 18(250 mm×4.6 mm,5μm)chromatography column was used,with a gradient elution of acetonitrile(A)-0.1%formic acid(B)as the mobile phase.The flow rate was 1.0 mL/min,the detection wavelength was 290 nm,the column temperature was 30℃,and the injection amount was 10μL.Using the similarity evaluation system for traditional Chinese medicine chromatography fingerprint(2012 version),HPLC fingerprint overlay spectra were established for 15 batches of C.behen roots and similarity was analyzed.By comparing and identifying the control samples,chemical components were confirmed,and content was determined.Hierarchical cluster analysis(HCA),principal component analysis(PCA),and orthogonal partial least squares discriminant analysis(OPLS-DA)were performed using SPSS 26.0 and SIMCA-P 14.1 softwares to evaluate the quality of different batches of thesamples.Results:HPLC fingerprint spectra of Fifteen batches of C.behen L.root was established,and a total of 20 common peaks were matched.Chlorogenic acid(peak11),caffeic acid(peak13)and isochlorogenic acid A(peak19)were identified by comparison of reference substance.A method for determining the content of chlorogenic acid and isochlorogenic acid A has been established;The similarity range of the fingerprint spectrum was 0.601~0.983.HCA divided 15 batches of C.behen into 2 categories;The cumulative variance contribution of the first 4 principal components derived from PCA was exactly 85.260%;The OPLS-DA showed that 2 components,including chlorogenic acid and isochlorogenic acid A,were the symbolic components of the quality difference of C.behen L.root.Conclusion:The combination of HPLC fingerprinting with chemical pattern recognition techniques allows for the rapid differentiation of characteristic components and effective evaluation of C.behen root.This integrated approach provides a robust basis for standardizing and ensuring the quality control of C.behen root.