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作者中文名	作者英文名	单位中文名	单位英文名	E-Mail
孙祎洋	sunyiyang	山西农业大学资源环境学院	College of Resources and Environment, Shanxi Agricultural University	x17516216330@163.com
饶之望	raozhiwang	山西农业大学资源环境学院	College of Resources and Environment, Shanxi Agricultural University	rzwww11@126.com
郭军玲	guojunling	1.山西农业大学资源环境学院，太原 030031 2.山西农业大学生态环境产业技术研究院/土壤环境与养分资源山西省重点实验室，太原 030031 3.山西农业大学农业农村部盐碱地改良与利用（干旱与半干旱盐碱地）重点实验室，太原 030031	1. College of Resources and Environment, Shanxi Agricultural University, Taiyuan 030031 2. Shanxi Agricultural University Ecological Environment Industry Technology Research Institute / Soil Shanxi Provincial Key Laboratory of Environment and Nutrient Resources, Taiyuan 030031 3. Shanxi Agricultural University, Ministry of Agriculture and Rural Saline-alkali Land Key Laboratory of Improvement and Utilization ( Arid and Semi-arid Saline-alkali Land ), Taiyuan 030031	guojunling6178@126.com
张强*	zhnagqiang	1.山西农业大学资源环境学院，太原 030031 2.山西农业大学生态环境产业技术研究院/土壤环境与养分资源山西省重点实验室，太原 030031 3.山西农业大学农业农村部盐碱地改良与利用（干旱与半干旱盐碱地）重点实验室，太原 030031	1. College of Resources and Environment, Shanxi Agricultural University, Taiyuan 030031 2. Shanxi Agricultural University Ecological Environment Industry Technology Research Institute / Soil Shanxi Provincial Key Laboratory of Environment and Nutrient Resources, Taiyuan 030031 3. Shanxi Agricultural University, Ministry of Agriculture and Rural Saline-alkali Land Key Laboratory of Improvement and Utilization ( Arid and Semi-arid Saline-alkali Land ), Taiyuan 030031	Zhangqiang0351@163.com
杨志平	yangzhiping	1.山西农业大学资源环境学院，太原 030031 2.山西农业大学生态环境产业技术研究院/土壤环境与养分资源山西省重点实验室，太原 030031 3.山西农业大学农业农村部盐碱地改良与利用（干旱与半干旱盐碱地）重点实验室，太原 030031	1. College of Resources and Environment, Shanxi Agricultural University, Taiyuan 030031 2. Shanxi Agricultural University Ecological Environment Industry Technology Research Institute / Soil Shanxi Provincial Key Laboratory of Environment and Nutrient Resources, Taiyuan 030031 3. Shanxi Agricultural University, Ministry of Agriculture and Rural Saline-alkali Land Key Laboratory of Improvement and Utilization ( Arid and Semi-arid Saline-alkali Land ), Taiyuan 030031	yzpsx0208@163.com

基金项目:山西省科技合作交流专项项目(202304041101042)；山西农业大学地方合作项目（XDHZHQY2022-02 ）；山西省农业科学院应用基础研究计划（YCX2020YQ15）

中文摘要:本研究采用超高效液相色谱-三重四极杆质谱联用仪（UPLC-TQ-MS），建立了一种适用于同时测定黄芪6种成分（黄芪甲苷、毛蕊异黄酮、毛蕊异黄酮苷、芒柄花素、芒柄花苷、芦丁）的方法，并探究了黄芪活性成分与根系形态特征的相关性。通过比较多个洗脱梯度及不同流速对峰形、分离度和质谱响应值的影响，最终确定采用乙腈-0.1%甲酸溶液为流动相，0.4 mL·min-1梯度洗脱效果最佳。采用Acquity UPLC BEH C18 色谱柱（50 mm×2.1 mm，1.7 μm），在ESI离子模式下进行质谱扫描。采用该方法对48株黄芪样品的活性成分含量进行测定，测量根长、根直径、根干重，进行相关性分析和逐步回归分析。结果：6种活性成分在各自浓度范围内表现出良好的线性关系（R2≥0.996），精密度RSD在1.18%～4.56%，重复性RSD在1.49%～4.75%，平均加样回收率97.30%～102.90%。该方法具有耗时短，重复性良好，易操作等优点，可为黄芪的鉴定和质量标准的完善提供科学依据。相关分析及逐步回归分析表明，恒山黄芪活性成分与根系形态指标相关，黄芪甲苷与根直径、根干重显著负相关，芦丁与根干重显著负相关。随着根直径增粗，黄芪甲苷含量呈降低变化，毛蕊异黄酮苷呈升高变化。可见，恒山黄芪根系指标可以作为评估其活性成分含量的参考依据。

中文关键词:UPLC-TQ-MS 黄芪活性成分方法学根系形态特征

Establishment of a UPLC-TQ-MS Method for Simultaneous Determination of Six Active Constituents in Astragalus membranaceus.

Abstract:In this study, an ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQ-MS ) was used to establish a method for simultaneous determination of six components (astragaloside IV, calycosin, calycosin-7-O-β-D-glucopyranoside, formononetin, ononin, rutin ) of Astragalus membranaceus, and to explore the correlation between active components and root morphological characteristics. By comparing the effects of multiple elution gradients and different flow rates on peak shape, resolution and mass spectrometry response values, it was finally determined that acetonitrile-0.1 % formic acid solution was used as the mobile phase, and 0.4 mL · min-1 gradient elution was the best. Acquity UPLC BEH C18 chromatographic column (100 mm × 2.1 mm, 1.7 μm ) was used for mass spectrometry scanning in ESI ion mode. The method was used to determine the content of active ingredients in 48 samples of Astragalus membranaceus. The root length, root diameter and root dry weight were measured, and correlation analysis and stepwise regression analysis were performed. The results demonstrated exceptional linear relationships within their respective concentration ranges (R2≥0.996), with precision RSD ranging from 1.18% to 4.56%, repeatability RSD from 1.49% to 4.75%, and average spiked recovery rates from 97.30% to 102.90%. This method has the advantages of short time-consuming, good repeatability and easy operation, which can provide scientific basis for the identification of Astragalus and the improvement of quality standards. Correlation analysis and stepwise regression analysis showed that the active components of A.hengshanense were correlated with root morphological indexes. Astragaloside IV was significantly negatively correlated with root diameter and root dry weight, and rutin was significantly negatively correlated with root dry weight. With the increase of root diameter, the content of astragaloside IV decreased and the content of calycosin-7-O-β-D-glucoside increased. It can be seen that the root index of Hengshan Astragalus can be used as a reference for evaluating the content of its active ingredients.

keywords:UPLC-TQ-MS Astragalus Active ingredients Methodology Root morphological characteristics

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