基于UPLC-Q TOF MS/MS技术结合SWATH采集法同时定性定量分析地胆草中的化学成分
UPLC-Q TOF MS/MS with SWATH method for simultaneous qualitative and quantitative analysis of chemical constituents in Elephantopi Herba
果佳慧 1蔡于罗 1田伟 2高乐 2李葆林 2甄亚钦 2牛丽颖2
作者信息
- 1. 河北中医药大学,石家庄 050091
- 2. 河北中医药大学,石家庄 050091;河北省中药配方颗粒技术创新中心,石家庄 050091;中药材品质评价与标准化河北省工程研究中心,石家庄 050091
- 折叠
摘要
目的:基于超高效液相色谱串联四极杆飞行时间质谱联用(UPLC-Q TOF MS/MS)技术,系统表征地胆草中化学成分并同时测定其中13个化学成分(新绿原酸、绿原酸、隐绿原酸、木犀草素-7-O-葡萄糖醛酸苷、木犀草苷、异绿原酸B、异绿原酸A、芹菜素-7-O-β-D-葡萄糖醛酸苷、异绿原酸C、去氧地胆草素、异去氧地胆草素、异地胆草种内酯、地胆草种内酯)的含量.方法:采用UPLC-Q TOF MS/MS技术,结合顺序窗口采集所有理论质谱模式(SWATH),采集不同批次地胆草样品的数据.使用Waters HSS T3 C18(100 mm×2.1 mm,1.8 μm)色谱柱,以乙腈(A)-0.1％甲酸水溶液(B)为流动相,梯度洗脱,流速0.4 mL·min-1,柱温35℃,进样量2 μL.采用电喷雾离子源(ESI),正、负离子模式下采集数据.结合对照品信息,参考相关文献完成对地胆草中化学成分的结构鉴定.同时建立地胆草中13个化学成分的含量测定方法并进行方法学考察.结果:地胆草中初步鉴定出36个化学成分,其中包括有机酸类20个,黄酮类7个,倍半萜内酯类6个,香豆素类1个及其他成分2个;含量测定的13个成分在测定浓度范围内线性关系良好,相关系数≥0.999 0,精密度、稳定性、重复性等均符合要求,平均加样回收率为94.9％～103.5％,RSD≤3.8％.不同批次地胆草所含化学成分基本相似,但13个成分的含量存在一定差异.结论:本研究建立的UPLC-Q TOF MS/MS方法,可同时对地胆草进行定性定量分析,方法灵敏准确,稳定性和重复性良好,可以为地胆草的质量控制及开发提供方法参考及数据支撑.
Abstract
Objective:To systematically characterize and simultaneous determine 13 chemical components(neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,luteolin-7-glucuronide,cynaroside,isochloro-genic acid B,isochlorogenic acid A,apigenin-7-glucuronide,isochlorogenic acid C,deoxyelephantopin,isode-oxyelephantopin,isoscabertopin and scabertopin)in Elephantopi Herba based on ultra performance liquid chroma-tography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q TOF MS/MS).Methods:UPLC-Q TOF MS/MS technology combined with sequential window acquisition of all theoretical mass spectra(SWATH)mode was used to collect data of different batches of Elephantopi Herba samples.The separation was performed on a Waters HSS T3 C18(100 mm ×2.1 mm,1.8 μm)column with mobile phase consisting of acetonitrile and 0.1％for-mic acid water solution.The flow rate of gradient elution was 0.4 mL·min-1,the column temperature was 35 ℃and the injection volume was 2 μL.The electrospray ionization(ESI)source was employed to collect data under both positive and negative ion modes.With reference to reference substances and relevant literatures,chemical com-ponents in Elephantopi Herba were identified.At the same time,the determination method for 13 chemical compo-nents in Elephantopi Herba was established and validated.Results:In this study,the cleavage patterns of different classes of compounds were summarized,and 36 chemical components were initially identified,including 20 organic acids,7 flavonoids,6 sesquiterpene lactones,1 coumarin and 2 other components.All of the analytes showed good linearity(r≥0.999 0)in the tested ranges.The precision,repeatability and stability of the method were good for the 13 components.The average recoveries were in the range of 94.9％-103.5％with relative standard deviations(RSDs)≤3.8％.The chemical compositions of different batches of Elephantopi Herba were basically similar,but there were certain differences in the contents of 13 components.Conclusion:In this study,comprehensive charac-terization of the chemical constituents and relative quantitative analysis of Elephantopi Herba are carried out,and a rapid and efficient qualitative analysis method of Elephantopi Herba is established by UPLC-Q TOF MS/MS,which provides a reference for the screening and rational exploitation of the medicinal substances of Elephantopi Herba,and methodological reference and data support for the quality control and development of Elephantopi Herba.
关键词
地胆草/超高效液相色谱-四极杆-飞行时间串联质谱/顺序窗口采集所有理论质谱模式/化学成分鉴定/含量测定/质量控制Key words
Elephantopi Herba/UPLC-Q TOF MS/MS/SWATH/chemical composition identification/content determination/quality control引用本文复制引用
基金项目
河北省省级科技计划(23372502D)
河北省中医药局科研项目(2023125)
出版年
2024
NETL
NSTL
万方数据