目的 建立鲜地黄Rehmannia glutinosa指纹图谱及多指标定性定量检测方法，为鲜地黄药材的质量评价提供科学依据。方法 采用HPLC-PDA法建立鲜地黄指纹图谱，烘干法测定水分，药材横断面图像面积比值采集测定菊花芯指数；运用《中药色谱指纹图谱相似度评价系统》（2012版）对指纹图谱进行相似度评价，联合SIMCA.14.1软件进行层次聚类分析（hierarchical cluster analysis，HCA）和正交偏最小二乘法-判别分析（orthogonal partial least-squares discriminant analysis，OPLS-DA）筛选鲜地黄栽培品和野生品的差异性指标成分，同时建立其中4种苷类成分的含量测定方法和2种成分的薄层色谱（TLC）法鉴别。结果 鲜地黄栽培品和野生品环烯醚萜苷部位（iridoid glycoside fraction，IGF）和苯乙醇苷部位（Phenylethanol glycoside fraction，PGF）指纹图谱均确定10个共有峰，分别以梓醇和毛蕊花糖苷为参照峰比较各共有峰的相对保留时间和相对峰面积，指纹图谱相似度评价结果均大于0.9。栽培和野生各15批次样品水分和菊花芯指数平均值分别为（75.75±0.04）%、（67.98±0.14）%和（37.87±0.19）%、（30.27±0.13）%。HCA结果显示，栽培品和野生品被分为2类，通过OPLS-DA分析，其中VIP值排名前10的指标成分分别为IGF-1、IGF-3（梓醇）、PGF-4（毛蕊花糖苷）、PGF-1、IGF-6（益母草苷）、IGF-9、IGF-8、PGF-2、IGF-10、IGF-4（地黄苷D），对区分二者的差异性贡献较大；建立了检测梓醇和毛蕊花糖苷的TLC鉴别方法，HPLC-PDA法测定其中4种苷类成分（梓醇、地黄苷D、地黄苷A、益母草苷）的含量，各成分的线性关系良好（r＞0.999 0），精密度、重复性和稳定性良好，回收率98.74%～102.51%，RSD均小于4%。结论 鲜地黄HPLC指纹图谱的构建及化学模式识别可以区分栽培品和野生品；水分和菊花芯指数栽培品大于野生品；建立的TLC鉴别法、HPLC-PDA含量测定法用于鲜地黄药材多种化学成分的定性、定量分析，简便、快速、准确，为完善鲜地黄药材质量评价体系提供参考。

Objective To establish the fingerprint of fresh Rehmannia Radix and multi-index qualitative and quantitative detection method,to provide scientific basis for comprehensive quality evaluation of fresh Rehmannia Radix.Methods HPLC method was used to establish fresh Rehmannia Radix fingerprint, content determination method and TLC identification method, and the water content and chrysanthemum core index were determined. The similarity evaluation system of TCM Chromatographic Fingerprint (2012 edition) was used to evaluate the corresponding chromatographic similarity. Hierarchical cluster analysis (HCA) and orthogonal Partial least squares discriminant analysis (OPLS-DA) were performed in conjunction with SIMCA.14.1 software. Results The fingerprints of each of Iridoid glycoside fraction (IGF) and Phenylethanol glycoside fraction (PGF) of fresh Rehmannia glutinosa contain ten common peaks. The relative retention time and relative peak area of common peaks were compared with catalpol and verminoside as reference peaks respectively. The similarity evaluation results of fingerprints were all greater than 0.9 except for a few of samples. The HCA results showed that cultivated products and wild products were divided into 2 categories. Through OPLS-DA analysis, ten peaks with VIP value greater than 0.8 were respectively IGF-1, IGF-3 (catalpol), PGF-4 (verbascoside), PGF-1, IGF-6 (ajugol), IGF-9, IGF-8, PGF-2, IGF-10, IGF-4 (rehmanidin D) peaks, which make great contribution to distinguish the difference between them . A TLC method for the identification of catalpa and verbascoside in fresh Rehmannia Radix was established. A HPLC-PDA method for the simultaneous determination of four glycosides in fresh Rehmannia Radix (catalpol, rehmancin D, rehmancin A, leonurin) was established. A HPLC-PDA method was established for the simultaneous determination of four glycosides in fresh Rehmannia Radix (catalpol, rehmannioside D, rehmannioside A, ajugol). The linear relationship of each component was good (r > 0.9990), and the precision, repeatability and stability were good. The recovery rates ranged from 98.74% to 102.51%, and RSD were all lower than 4%. Conclusion The construction of HPLC fingerprint and chemical pattern recognition of fresh Rehmannia Radix could distinguish wild and cultivated products. The established TLC identification method and HPLC-PDA content determination method are simple, fast and accurate for the qualitative and quantitative analysis of various chemical components in fresh Rehmannia Radix.

R286.2

河南省中医药科学研究专项重大课题（20-21ZYZD13）;河南省中医药科学研究专项重点课题（2024ZY1026，2023ZY1023）;河南省省属科研机构基本科研业务费项目（2404024，2204966）