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、α-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、α-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（α-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。α-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α-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。

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, α-cyperone. Results The 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, α-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 (α-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 α-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 α-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.

R283.6

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）