目的 建立鮮益母草藥材指標成分含量及指紋圖譜測定方法，結合藥材安全性指標，綜合評價不同產(chǎn)地、不同采收期鮮益母草藥材的整體質量。方法 收集不同產(chǎn)地23批鮮益母草藥材，測定各批次藥材的UPLC指紋圖譜、鹽酸水蘇堿和鹽酸益母草堿的含量、5種重金屬及有害元素的含量以及33種禁用農(nóng)藥殘留量，采用主成分分析（PCA）、聚類分析（HCA）以及正交偏最小二乘法判別分析（OPLS-DA）等化學模式識別方法與熵權TOPSIS法對采集的各指標數(shù)據(jù)進行多元分析；同時收集3個產(chǎn)地不同采收期的鮮益母草藥材，測定各批次樣品的UPLC指紋圖譜、鹽酸水蘇堿和鹽酸益母草堿的含量，分析不同采收期鮮益母草化學成分的變化規(guī)律。結果 建立的鮮益母草藥材指紋圖譜共標定15個峰，指認了6個峰，分別為峰2（新綠原酸）、峰4（綠原酸）、峰8（鹽酸益母草堿）、峰12（蘆?。?、峰13（異槲皮苷）、峰15（椴樹苷）；不同批次鮮益母草藥材指紋圖譜與指標成分的含量存在一定差異，23批鮮益母草藥材與對照指紋圖譜的相似度＞0.9，鹽酸益母草堿含量為0.17～0.78 mg·g^-1，鹽酸水蘇堿含量為0.71～5.82 mg·g^-1；鉛、鎘、砷、汞、銅的含量均符合《中國藥典》對重金屬及有害元素的一般限量要求，33種禁用農(nóng)藥殘留量均未檢出。PCA和HCA均可將23批鮮益母草藥材大致聚為3類，采收期對鮮益母草藥材的整體質量影響較大；OPLS-DA篩選出鹽酸益母草堿的含量及峰1、峰5、峰7、峰8的峰面積是影響鮮益母草質量差異的主要因素；熵權TOPSIS分析結果顯示，幼苗期的鮮益母草藥材的整體質量較優(yōu)；4～7月份采收的鮮益母草中鹽酸益母草堿、鹽酸水蘇堿含量及峰2～5、峰10～12、峰14的峰面積隨時間延長均呈現(xiàn)下降趨勢，峰6、7、9、13、15的峰面積先上升后下降，峰1的峰面積則呈現(xiàn)波動狀態(tài)。結論 建立的鮮益母草藥材指紋圖譜和指標成分含量測定方法，精密度高，重復性好，結果穩(wěn)定可靠，綜合質量評價結果及不同采收期化學成分變化規(guī)律可為鮮益母草的采收及質量控制提供指導。

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

廣東省中醫(yī)藥研究開發(fā)重點實驗室開放基金資助項目（KFKT02-007）;廣東省中醫(yī)藥局科研項目（科研平臺專項）（20233006）