目的 建立鲜益母草药材指标成分含量及指纹图谱测定方法，结合药材安全性指标，综合评价不同产地、不同采收期鲜益母草药材的整体质量。方法 收集不同产地23批鲜益母草药材，测定各批次药材的UPLC指纹图谱、盐酸水苏碱和盐酸益母草碱的含量、5种重金属及有害元素的含量以及33种禁用农药残留量，采用主成分分析（PCA）、聚类分析（HCA）以及正交偏最小二乘法判别分析（OPLS-DA）等化学模式识别方法与熵权TOPSIS法对采集的各指标数据进行多元分析；同时收集3个产地不同采收期的鲜益母草药材，测定各批次样品的UPLC指纹图谱、盐酸水苏碱和盐酸益母草碱的含量，分析不同采收期鲜益母草化学成分的变化规律。结果 建立的鲜益母草药材指纹图谱共标定15个峰，指认了6个峰，分别为峰2（新绿原酸）、峰4（绿原酸）、峰8（盐酸益母草碱）、峰12（芦丁）、峰13（异槲皮苷）、峰15（椴树苷）；不同批次鲜益母草药材指纹图谱与指标成分的含量存在一定差异，23批鲜益母草药材与对照指纹图谱的相似度＞0.9，盐酸益母草碱含量为0.17～0.78 mg·g^-1，盐酸水苏碱含量为0.71～5.82 mg·g^-1；铅、镉、砷、汞、铜的含量均符合《中国药典》对重金属及有害元素的一般限量要求，33种禁用农药残留量均未检出。PCA和HCA均可将23批鲜益母草药材大致聚为3类，采收期对鲜益母草药材的整体质量影响较大；OPLS-DA筛选出盐酸益母草碱的含量及峰1、峰5、峰7、峰8的峰面积是影响鲜益母草质量差异的主要因素；熵权TOPSIS分析结果显示，幼苗期的鲜益母草药材的整体质量较优；4～7月份采收的鲜益母草中盐酸益母草碱、盐酸水苏碱含量及峰2～5、峰10～12、峰14的峰面积随时间延长均呈现下降趋势，峰6、7、9、13、15的峰面积先上升后下降，峰1的峰面积则呈现波动状态。结论 建立的鲜益母草药材指纹图谱和指标成分含量测定方法，精密度高，重复性好，结果稳定可靠，综合质量评价结果及不同采收期化学成分变化规律可为鲜益母草的采收及质量控制提供指导。

Objective To establish a method for the determination of the index components and fingerprint of fresh Leonurus japonicus, and to comprehensively evaluate the overall quality of fresh L. japonicus from different origins and different harvest dates in combination with the drug safety index. Method A total of 23 batches of fresh L. japonicus in different regions were collected, the fingerprints, stachydrine hydrochloride content, 5 kinds of leonurine hydrochloride content, heavy metals and harmful elements content and 33 prohibited pesticide residues of each batch were determined, and the data of each index component were analyzed by chemical pattern recognition such as principal component analysis (PCA), hierarchical cluster analysis (HCA), orthogonal partial least squares discriminant analysis (OPLS-DA) and entropy weight TOPSIS method, at the same time, fresh L. japonicus from three regions at different harvest times were collected, and determined the fingerprints, stachydrine hydrochloride content and leonurine hydrochloride content, and analyzed the changes of chemical components of fresh L. japonicus. Result The established fingerprint spectrum of fresh L. japonicus was calibrated with 15 peaks, and 6 peaks were identified, which were peak 2 (neochlorogenic acid), peak 4 (chlorogenic acid), peak 8 (leonurus hydrochloride), peak 12 (rutin), peak 13 (isoquercetin), and peak 15 (tiliroside). There were some differences in fingerprints and content of index components of different batches of fresh L. japonicus, the similarity between the 23 batches of fresh L. japonicus and the control characteristic chromatography was greater than 0.9, the fluctuation range of leonurine hydrochloride content was 0.17 to 0.78 mg·g^-1, and the fluctuation range of stachydrine hydrochloride content was 0.71 to 5.82 mg·g^-1, the contents of lead, cadmium, arsenic, mercury and copper met the general limit requirements for heavy metals and harmful elements in the Pharmacopoeia, and 33 prohibited pesticide residues were not detected. PCA and HCA could aggregate 23 batches of fresh L. japonicus into three categories, and the harvest time had a great impact on the overall quality of fresh L. japonicus, OPLS-DA screened out that the content of leonurine hydrochloride and the peak areas of peaks 1, 5, 7, and 8 the main factors affecting the quality of fresh L. japonicus. The entropy weight TOPSIS method showed that the quality of fresh L. japonicus at seedling stage was better, the content of leonurine hydrochloride stachydrine hydrochloride and the peak areas of peak 2-5, peak 10-12 and peak 14 of fresh L. japonicus at different harvest times showed a downward trend, the peak areas of peak 6, 7, 9, 13 and 15 first increased and then decreased, and the peak area of peak 1 showed a fluctuating trend.Conclusion The method of establishing the fingerprint and content determination method of fresh L. japonicus was highly precise, reproducible and the results were stable and reliable, the results of comprehensive quality evaluation and the variation of chemical composition in different harvest periods can provide a reference for the comprehensive evaluation and control of the quality of fresh L. japonicus.

R927.2

广东省中医药研究开发重点实验室开放基金资助项目（KFKT02-007）;广东省中医药局科研项目（科研平台专项）（20233006）