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指纹图谱结合一测多评法评价山楂叶质量
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| 篇名： | 指纹图谱结合一测多评法评价山楂叶质量 |
| TITLE： | Quality evaluation of Crataegus pinnatifida leaves by fingerprint combined with quantitative analysis of multi-components by single-marker |
| 摘要： | 目的 建立不同产地山楂叶药材的指纹图谱和多成分含量测定方法，评价其质量并筛选差异标志物。方法收集产自河北省承德市、辽宁省葫芦岛市、山西省运城市、山东省临沂市的78批山楂叶药材，采用高效液相色谱（HPLC）法，利用《中药色谱指纹图谱相似度评价系统（2012版）》绘制指纹图谱并进行相似度评价；采用SPSS19.0、MetaboAnalyst5.0工具、SIMCA14.1软件等进行灰色关联分析、聚类分析（CA）、主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA），以变量重要性投影（VIP）值＞1且误差线不超过原点为标准筛选影响山楂叶质量的差异标志物。采用同一HPLC法结合一测多评（QAMS）法，以牡荆素鼠李糖苷为内参物，测定78批山楂叶药材中绿原酸、牡荆素葡萄糖苷、金丝桃苷和异槲皮素的含量，并与外标法测定结果进行比较。结果4个产地的78批山楂叶药材指纹图谱中有8个共有峰被标定，有5种成分[绿原酸（峰1）、牡荆素葡萄糖苷（峰3）、牡荆素鼠李糖苷（峰4）、金丝桃苷（峰7）和异槲皮素（峰8）]被指认，相似度为0.871～0.998。河北省承德市、辽宁省葫芦岛市、山西省运城市、山东省临沂市样品的平均相对关联度分别为0.538、0.528、0.462、0.435；CA、PCA分析均显示，河北省承德市和辽宁省葫芦岛市样品大致分为一类，山东省临沂市和山西省运城市样品大致分为一类；峰1、2、3、5的VIP值均大于1。QAMS法计算得绿原酸、牡荆素葡萄糖苷、金丝桃苷、异槲皮素的平均相对校正因子分别为0.401、0.993、1.670、1.615（RSD＜2%）；与外标法结果比较，除2批样品（S39、S41）中的异槲皮素外，其余批次样品中各成分的含量均无明显差异（相对偏差不高于5%）。结论所建指纹图谱和QAMS法操作简单，可用于评价山楂叶药材的质量；河北省承德市样品质量较优；绿原酸（峰1）、牡荆素葡萄糖苷（峰3）和峰2、5对应成分可能是影响山楂叶药材质量的差异标志物。 |
| ABSTRACT： | OBJECTIVE To establish the fingerprint and multi-component content determination method of Crataegus pinnatifida leaves from different producing areas， and to evaluate the quality of C. pinnatifida leaves and screen the differential markers. METHODS Seventy-eight batches of C. pinnatifida leaves were collected from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province. High-performance liquid chromatography （HPLC） and Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints （2012 edition） were used to draw the fingerprints and conduct similarity evaluation. Grey correlation analysis， cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were performed by using SPSS 19.0， MetaboAnalyst 5.0 and SIMCA 14.1 software. The differential markers affecting the quality of C. pinnatifida leaves were screened with variable importance in the projection （VIP） value greater than 1 and the error line not exceeding the origin as the criterion. Using vitexin rhamnoside as an internal reference， the contents of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin in 78 batches of C. pinnatifida leaves were determined by the same HPLC combined with quantitative analysis of multi- components by single-marker （QAMS）， and the results were compared with external standard method. RESULTS Eight common peaks were calibrated in the fingerprints for 78 batches of C. pinnatifida leaves from 4 producing areas. Five known components were identified， including chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， vitexin rhamnoside （peak 4）， hypericin （peak 7） and isoquercetin （peak 8）； their similarities ranged from 0.871 to 0.998. Average relative correlations of samples from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province were 0.538， 0.528， 0.462 and 0.435， respectively. CA and PCA showed that the samples from Chengde of Hebei Province and Huludao of Liaoning Province were roughly classified into one category， while the samples from Linyi of Shandong Province and Yuncheng of Shanxi Province were roughly classified into one category； VIP values of peak 1， 2， 3 and 5 were all greater than 1. By QAMS， the relative correction factors of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin were 0.401， 0.993， 1.670 and 1.615 （RSD＜2%）. Compared with external standard method， except for isoquercetin in the two batches of samples （S39 and S41）， there was no significant difference in the content of each component in other batches of samples （the relative deviations≤ 5%）. CONCLUSIONS The established fingerprint and QAMS method are simple to operate and can be used to evaluate the quality of C. pinnatifida leaves. The sample from Chengde of Hebei Province is relatively good in quality. Chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， and the corresponding components of peaks 2 and 5 may be differential markers affecting the quality of C. pinnatifida leaves. |
| 期刊： | 2023年第34卷第22期 |
| 作者： | 李艳荣；段丽颖；魏红；杜义龙；赵胜男；高晗；潘海峰 |
| AUTHORS： | LI Yanrong，DUAN Liying，WEI Hong，DU Yilong，ZHAO Shengnan，GAO Han，PAN Haifeng |
| 关键字： | 山楂叶；指纹图谱；一测多评法；化学模式识别；差异标志物 |
| KEYWORDS： | Crataegus pinnatifida leaves； fingerprints； quantitative analysis of multi-components by single-marker； chemical |
| 阅读数： | 55 次 |
| 本月下载数： | 1 次 |
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