基于化学模式识别筛选清炒香附的差异标志物_杨颜溶.pdf

中草药 中草药 2023 年年 3 月月 第第 54 卷卷 第第 5 期期 Chinese Traditional and Herbal Drugs 2023 March Vol.54 No.5 1411 基于化学模式识别筛选清炒香附的差异标志物 杨颜溶1,2，田瀚举1,2，李莹莹1,2，贾 豪1,2，雷敬卫1,2*，龚海燕1,2*，谢彩侠1,2 1.河南中医药大学，河南 郑州 450046 2.河南省中药质量控制与评价工程技术中心，河南 郑州 450046 摘 要：目的 建立香附饮片、清炒香附、香附炭的 HPLC 指纹图谱，结合化学模式识别筛选差异标志物，并测定 6 种主要成分的含量，为进一步完善香附质量标准提供参考。方法 采用中药色谱指纹图谱相似度评价系统（2012 版）建立 21 批香附样品的 HPLC 指纹图谱，进行相似度评价，确定共有峰个数；采用 SIMCA 14.1 软件进行层次聚类分析（hierarchical cluster analysis，HCA）、主成分分析（principal component analysis，PCA）、正交偏最小二乘-判别分析（orthogonal partial least squares-discriminant analysis，OPLS-DA），以变量重要性投影（variable importance in projection，VIP）1 为标准筛选出差异标志物；测定 5-羟基甲基糠醛（5-hydroxymethylfurfural，5-HMF）、对香豆酸、阿魏酸、木犀草素、香附烯酮、-香附酮的含量。结果 21 批 HPLC 指纹图谱的相似度均不小于 0.934，共标定 23 个共有峰，指认 3、10、11、17、21、23 号峰分别为 5-HMF、对香豆酸、阿魏酸、木犀草素、香附烯酮、-香附酮；PCA 结果、OPLS-DA 结果与 HCA 结果一致，均可将 21 批香附样品分为 3 类；通过 VIP 筛选出 2、23（-香附酮）、17（木犀草素）、10（对香豆酸）、11（阿魏酸）号峰为导致样品差异的主要标志色谱峰；5-HMF 的含量在香附炭中最高。对香豆酸、阿魏酸的含量在香附饮片、清炒香附、香附炭中逐渐增加。木犀草素在清炒香附中含量最高。-香附酮含量经炒后略有增加。香附烯酮含量在香附饮片、清炒香附、香附炭中含量逐渐减少。结论 筛选出未知成分 2 号峰与已知成分-香附酮、木犀草素、对香豆酸、阿魏酸为差异标志物；所建立的清炒香附的 HPLC 指纹图谱存在一定差异，且 HPLC 指纹图谱与含量测定方法操作简单、准确，能够为清炒香附药效物质基础研究提供实验依据，可为进一步完善香附质量标准提供参考。关键词：香附；清炒；指纹图谱；含量；化学模式识别；差异标志物；层次聚类分析；主成分分析；正交偏最小二乘-判别分析；5-羟基甲基糠醛；对香豆酸；阿魏酸；木犀草素；香附烯酮；-香附酮 中图分类号：R283.6 文献标志码：A 文章编号：0253-2670(2023)05-1411-08 DOI:10.7501/j.issn.0253-2670.2023.05.007 Screening difference markers of stir-fried Cyperi Rhizoma based on chemical pattern recognition YANG Yan-rong1,2,TIAN Han-ju1,2,LI Ying-ying1,2,JIA Hao1,2,LEI Jing-we1,2,GONG Hai-yan1,2,XIE Cai-xia1,2 1.Henan University of Traditional Chinese Medicine,Zhengzhou 450046,China 2.Henan Provincial Chinese Medicine Quality Control and Evaluation Engineering Technology Center,Zhengzhou 450046,China Abstract:Objective To establish the HPLC fingerprint of Xiangfu(Cyperi Rhizoma)decoction pieces,stir-fried Cyperi Rhizoma,and Cyperi Rhizoma charcoal,combine chemical pattern recognition to screen different markers,and determine the content of six main ingredients,providing a reference for further improving the quality standards of Cyperi Rhizoma.Methods The HPLC fingerprints of 21 batches of Cyperi Rhizoma samples were established by the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 Edition),and the similarity evaluation was carried out to determine the total number of peaks.SIMCA 14.1 software was used for hierarchical cluster analysis(HCA),principal component analysis(PCA),orthogonal partial least squares-discriminant analysis(OPLS-DA),and VIP 1 was used as the criterion to screen out the difference markers;Determination of the content of 5-hydroxymethylfurfural(5-HMF),p-coumaric acid,ferulic acid,luteolin,caryophyllone,-cyperone.Results The 收稿日期：2022-10-08 基金项目：国家重点研发计划项目（2018YFC1707000）；河南省中医药科学研究专项（20-21ZY2095）作者简介：杨颜溶（1998），硕士研究生，研究方向为中药质量分析研究。Tel:15090522158 E-mail:*通信作者：雷敬卫，教授，硕士生导师，博士，从事中药质量分析研究。Tel:(0371)656955281 E-mail: 龚海燕，副教授，硕士研究生，从事中药质量分析研究。Tel:13838393243 E-mail:ghy_ 1412 中草药 中草药 2023 年年 3 月月 第第 54 卷卷 第第 5 期期 Chinese Traditional and Herbal Drugs 2023 March Vol.54 No.5 similarity of the fingerprints of the 21 batches of HPLC was not less than 0.934,and a total of 23 common peaks were calibrated,and peaks 3,10,11,17,21 and 23 were identified as 5-HMF,p-coumaric acid,ferulic acid,luteolin,caryophyllone,-cyperone,respectively.The results of PCA,OPLS-DA results were consistent with the results of HCA,and 21 batches of Cyperi Rhizoma samples can be divided into three categories;By VIP method,2,23(-cyperone),17(luteolin),10(p-coumaric acid),11(ferulic acid)were selected as the main marker chromatographic peaks that caused the difference in samples;The content of 5-HMF was highest in Cyperi Rhizoma charcoal.The content of coumaric acid and ferulic acid gradually increased in Cyperi Rhizoma decoction pieces,stir-fried Cyperi Rhizoma and Cyperi Rhizoma charcoal.Luteolin was the highest in stir-fried Cyperi Rhizoma.The content of-cyperone increased slightly after stir-frying.The content of caryophyllone was gradually reduced in Cyperi Rhizoma decoction pieces,stir-fried Cyperi Rhizoma,and Cyperi Rhizoma charcoal.Conclusion The unknown ingredient peak 2 was screened and the known ingredients-cyperone,luteolin,p-coumaric acid and ferulic acid were selected as markers of difference;There were certain differences in the HPLC fingerprint of the fried Cyperi Rhizoma,and the HPLC fingerprint and content determination method were simple and accurate to operate,which can provide experimental basis for the basic research of the medicinal substances attached to the stir-fried Cyperi Rhizoma,and provide a reference for further improving the quality standards of the Cyperi Rhizoma.Key words:Cyperi Rhizoma;stir-frying;fingerprinting;content;chemical pattern recognition;difference markers;hierarchical cluster analysis;principal component analysis;orthogonal partial least squares-discriminant analysis;5-hydroxymethylfurfural;p-coumaric acid;ferulic acid;luteolin;caryophyllone;-cyperone 香附最早记载于 名