HPLC指纹图谱和多成分定量结合化学模式识别评价菊苣子质量

Quality evaluation of chicory seed based on HPLC fingerprint and multi-component quantification combined with chemical pattern recognition

目的建立腺毛菊苣Cichorium glandulosum种子(菊苣子药材)和混伪品菊苣Cichorium intybus种子HPLC指纹图谱及多成分含量测定,并结合化学模式识别法对二者进行分析比较,为其资源利用开发和进一步的质量控制研究提供参考。方法采用YMC-PACK ODS-A色谱柱,以0.2%磷酸溶液(A)-甲醇(B)为流动相梯度洗脱,体积流量为1.0 mL/min,检测波长为254 nm,柱温为40℃,进样量为5μL。利用《中药色谱指纹图谱相似度评价系统(2012版)》建立19批腺毛菊苣种子和4批菊苣种子样品的HPLC指纹叠加图谱并分析相似度,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 20.0和SIMCA-P 14.1软件进行聚类分析、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19批腺毛菊苣种子和4批菊苣种子HPLC指纹叠加图谱,并确定19个共有峰,通过对照品比对指认了其中8个色谱峰;建立了绿原酸、秦皮乙素、1,4-二咖啡酰奎宁酸、异绿原酸A、1,5-二咖啡酰奎宁酸含量测定方法;指纹图谱相似度在0.915~0.998;OPLS-DA分析与PCA分析结果一致,可将样品分为3组,并筛选出1,5-二咖啡酰奎宁酸、槲皮素、绿原酸、菊苣酸4个差异性成分。结论通过HPLC指纹图谱结合化学模式识别方法,建立了一种简单可靠的菊苣子药材质量评价方法,为菊苣子药材的质量评价和资源化利用提供依据。

Objective To establish HPLC fingerprint spectra and multi-component content determination of Cichorium glandulosum seeds(chicory seed medicinal material)and counterfeit C.intybus seeds,and analyze and compare the two using chemical pattern recognition method,providing reference for their resource utilization and further quality control research.Methods YMC-PACK ODS-A chromatography column was used,with a gradient elution of 0.2% phosphoric acid solution(A)-methanol(B)as the mobile phase.The flow rate was 1.0 mL/min,the detection wavelength was 254 nm,the column temperature was 40℃,and the injection amount was 5μL.Using the similarity evaluation system for traditional Chinese medicine chromatography fingerprint(2012 version),HPLC fingerprint overlay spectra were established for 19 batches of C.glandulosum seeds and four batches of C.intybus seeds samples,and similarity was analyzed.By comparing and identifying the control samples,chemical components were confirmed and content was determined.Cluster analysis,principal component analysis(PCA),and orthogonal partial least squares discriminant analysis(OPLS-DA)were performed using SPSS 20.0 and SIMCA-P 14.1 softwares to evaluate the quality of different batches of samples.Results A HPLC fingerprint overlay map was established for19 batches of C.glandulosum seeds and four batches of C.intybusseeds,and 19 common peaks were identified.A total of eight chromatographic peaks were identified through comparison with the control samples;A method for determining the content of chlorogenic acid,aesculetin,1,4-dicaffeoyl quinic acid,isochlorogenic acid A,and 1,5-dicaffeoyl quinic acid had been established;The similarity of fingerprint spectra was between 0.915 and 0.998;The results of OPLS-DA were consistent with those of PCA.The samples can be divided into three groups and four differential components,including 1,5-dicaffeoylquinic acid,quercetin,chlorogenic acid,and cichoric acid,can be selected.Conclusion A simple and reliable quality evaluation method for chicory seed medicinal material was established by combining HPLC fingerprint with chemical pattern recognition,providing a basis for the quality evaluation and resource utilization of chicory seed medicinal material.