摘要
目的:建立一种基于高效液相色谱串联四极杆轨道阱质谱(HPLC-QExactive MS)的分析方法,对补阳还五汤中的化学成分进行快速鉴定,并以高效液相色谱紫外检测(HPLC-UV)对其中含量较高的成分进行定量检测。方法:通过文献检索建立补阳还五汤相关化合物数据库;使用HPLC-Q-Exactive MS系统在全扫描/数据依赖性二级质谱(dd-MS^(2))负离子模式下采集质谱信息;借助Compound Discoverer 3.0软件分析推断补阳还五汤中的化学成分;进一步建立HPLC-UV含量测定方法,对其芍药苷、没食子酸、羟基红花黄色素A、阿魏酸、毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮、芒柄花素等8种活性成分进行定量分析。结果:通过定性分析,共鉴定出补阳还五汤中178个化合物。HPLC-UV定量分析方法学考察结果显示,8个主要成分在各自浓度范围内线性关系良好(R^(2)≥0.9990),精密度和稳定性相对标准偏差(RSD)均小于15%,加样回收率RSD为1.6%~2.4%。结论:本研究所建立的方法体系实现了补阳还五汤中成分的快速识别和主要成分的精确定量,其中芍药苷、羟基红花黄色素A和没食子酸含量较高,有望作为质控标志物。
Objective:To establish an analytical method for rapid identification of chemical compositions and quantitative determination of major compositions in Buyang Huanwu decoction(BYHWD)based on high performance liquid chromatographyquadrupole orbitrap mass spectrometry(HPLC-Q-Exactive MS)and high performance liquid chromatography-ultraviolet detection(HPLC-UV).Methods:The mass spectrometry information was collected in Full MS/dd-MS^(2) negative ion mode with HPLC-QExactive MS system;the chemical compositions of BYHWD were subsequently annotated with Compound Discoverer 3.0 software and a self-built in-house compound library.Eight major compositions(paeoniflorin,gallic acid,hydroxysafflor yellow A,ferulic acid,calycosin-7-glucoside,ononin,calycosin,formononetin)were picked out and their contents were quantitatively determined with HPLC-UV analysis.Results:A total of 178 compounds in BYHWD were tentatively identified.The results of HPLC-UV quantitative analysis showed that 8 compositions had a good linear relationship in their respective concentration range(R^(2)≥0.9990),the relative standard deviations(RSD)of precision and stability were all less than 15%,and the recovery rate RSD was between 1.6%and 2.4%.Conclusions:The method established in this study can realize the rapid identification and accurate quantification of the major compositions in BYHWD.Paeoniflorin,hydroxysafflor yellow A and gallic acid may be used as quality control markers.
作者
郑留燕
万浩芳
鲁一航
丁志山
李畅
万海同
ZHENG Liuyan;WAN Haofang;LU Yihang;DING Zhishan;LI Chang;WAN Haitong(Zhejiang Provincial TCM Key Laboratory of Encephalopathy,Zhejiang Chinese Medical University,Hangzhou 310053,China)
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2022年第5期534-543,共10页
Journal of Zhejiang University(Medical Sciences)
基金
国家重点研发计划(2019YFC1708600,2019YFC1708604)
浙江省中医药科技计划(2021ZQ025)
浙江省自然科学基金(LR19H280001)。
