基于质量标志物有效性、特有性及可测性理念的天葵子Q-marker成分预测  

Q-marker component prediction of Semiaquilegiae Radix based on the concept of effectiveness,specificity and testability of quality markers

作　　者：陈莉[1] 王盛[1] 刘柳珊 杨俊[2] 孟楣[1] 詹志来 CHEN Li;WANG Sheng;LIU Liu-shan;YANG Jun;MENG Mei;ZHAN Zhi-lai(The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China;Anhui University of Chinese Medicine,Hefei 230031,China;State Key Laboratory of Dao-di Herbs Breeding Base,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]安徽中医药大学第一附属医院,合肥230031 [2]安徽中医药大学,合肥230031 [3]中国中医科学院中药资源中心道地药材国家重点实验室培育基地,北京100700

出　　处：《药物分析杂志》2023年第10期1672-1683,共12页Chinese Journal of Pharmaceutical Analysis

基　　金：中央本级重大增减支项目,名贵中药资源可持续利用能力建设项目,天葵子、儿茶和败酱草药材商品规格等级标准研制(2060302-1906-23);全国中医药特色技术传承人才培训项目(T20184828005)。

摘　　要：目的:桥连网络药理学和特征图谱技术,预测天葵子质量标志物,为天葵子全面的质量控制体系建立提供参考。方法:文献及成分-靶点-通路网络可视化研究,定位并预测天葵子干预疾病的潜在质量标志物;采用Agilent Eclipse Plus C18(250 mm×4.6 mm,5μm)色谱柱,柱温30℃,以乙腈(A)-含15%乙腈的0.1%磷酸水溶液(每100 mL含SDS 0.2 g)(B)为流动相,梯度洗脱(0~5 min,0%A;5~10 min,0%A→27%A;10~33 min,27%A→29%A;33~38 min,29%A→37%A;38~55 min,37%A→38%A;55~72 min,38%A),流速0.8 mL·min^(-1),检测波长为260 nm(0~6 min)、220 nm(6~10 min)、260 nm(10~29 min)、220 nm(29~34 min)、260 nm(34~37 min)、220 nm(37~72 min);对收集的38批天葵子药材进行HPLC特征图谱定性定量分析,运用OPLS-DA筛选出造成组间差异的主要标志性成分;根据质量标志物的有效性、特有性及可测性预测天葵子潜在的Q-marker。结果:锁定了天葵子质量标志物的主要来源范围,网络药理学预测了其干预疾病的3个潜在质量标志物;成功建立了7个共有峰的特征图谱,标定了4个特征峰,其中3个差异性代谢产物,且格列风内酯、木兰花碱和小檗红碱质量浓度分别在2.22~44.4、14.2~284、3.8~76μg·mL^(-1)范围内均与峰面积积分值呈良好的线性关系,平均加样回收率分别为98.8%、98.8%和99.6%,RSD分别为0.64%、1.7%和1.9%;38批样品中格列风内酯、木兰花碱和小檗红碱的含量范围分别为0.046~0.757、0.391~2.796、0.077~0.572 mg·g^(-1)。根据质量标志物的有效性、特有性及可测性,预测天葵子中格列风内酯和小檗红碱2个成分为天葵子的Q-marker成分。结论:本研究预测了天葵子的Q-marker成分,同时建立了简便、准确、可靠的天葵子特征图谱定性定量方法,为天葵子全面质量控制体系的建立提供了参考,研究结果将有利于市场上天葵子药材质量评价。Objective:To establish a comprehensive quality control system for Semiaquilegiae Radix.To predict the quality markers of Semiaquilegiae Radix by a bridged network pharmacology and feature map technology.Methods:Based on literature research and network pharmacology research,potential quality biomarkers of Semiaquilegiae Radix were predicted.Qualitative and quantitative analysis of HPLC characteristic chromatogram was carried out on 38 batches of Semiaquilegiae Radix.Agilent Eclipse Plus C_(18)chromatographic column(250 mm×4.6 mm,5μm)was used.The mobile phase was acetonitrile(A)and a 0.1%phosphoric acid aqueous solution containing 15%acetonitrile(containing 0.2 g of SDS per 100 mL)(B),with gradient elution(0-5 min,0%A;5-10 min,0%A→27%A;10-33 min,27%A→29%A;33-38 min,29%A→37%A;38-55 min,37%A→38%A;55-72 min,38%A)at flow rate of 0.8 mL·min~(-1).The column temperature was 30℃,and the detection wavelengths were 260 nm for 0-6 min,220 nm for 6-10 min,260 nm for 10-29 min,220 nm for 29-34 min,260 nm for 34-37 min,and 220 nm for 37-72 min.OPLS-DA was used to screen the main marker components causing the difference between groups.According to the effectiveness,specificity and testability of quality markers,the potential Q-marker of Semiaquilegiae Radix was predicted.Results:The main source range of quality markers of Semiaquilegiae Radix was locked,and three potential quality markers for disease intervention were predicted by network pharmacology.The characteristic chromatogram with 7 common peaks was successfully established,and 3 differential metabolites were screened.Contents of glifepristone,magnoflorine,and berberine were between 2.22 and 44.4,respectivelyμg·mL^(-1),14.2-284μg·mL^(-1)and 3.8-76μg·mL^(-1).There was a good linear relationship between the concentration range and the peak area integral value in each of them,with average sample recovery rates of 98.8%,98.8%,and 99.6%,and RSD of 0.64%,1.7%,and 1.9%,respectively.The content ranges of glifepristone,magnoflorine,and berberine in 38 batches of

关 键 词：天葵子 网络药理学 HPLC特征图谱 多元统计分析 质量标志物 格列风内酯 小檗红碱 木兰花碱 定性定量研究 

分 类 号：R917[医药卫生—药物分析学]