基于UPLC-QTOF-MS/MS分析的芪葵颗粒化学成分系统筛选与识别

Systematic Screening and Identification of Chemical Constituents in Qikui Granules Based on UPLC-QTOF-MS/MS Analysis

目的系统定性表征芪葵颗粒(黄芪、制首乌、黄蜀葵花)中的化学成分。方法采用超高效液相色谱-四级杆飞行时间质谱(UPLC-QTOF-MS/MS)联用分析方法对芪葵颗粒水提液进行分离检测,获取正、负离子模式下的原始分析数据;借助AB Sciex PeakView软件,综合采用靶向和非靶向的筛选方法,对获得的原始分析数据进行处理,系统筛选鉴定芪葵颗粒中的化学成分;以不少于2个准分子离子的同时出现,排除碎片离子的干扰,锁定筛选所得目标化合物;依据准分子离子及特征二级子离子质谱信息,结合标准品比对,并参考数据库搜索匹配和文献数据比对,对锁定的化合物进行鉴定或合理归属。结果从芪葵颗粒中共筛选出89个化合物,鉴定出71个化合物,包括黄酮类45个,皂苷类14个,蒽醌类4个,二苯乙烯苷类3个,多糖类2个,有机酸类3个。鉴定结果表明羟基化、糖基化、葡萄糖醛酸化、乙酰化、甲基化、丙二酰化等多种转化反应参与了芪葵颗粒多样化学成分的形成。结论基于UPLC-QTOF-MS/MS靶向和非靶向相结合的筛选鉴定方法可快速全面表征芪葵颗粒中的主要化学成分及其涉及的转化反应,为深入探讨芪葵颗粒的质量控制、组方优化和药效物质等奠定了基础。

OBJECTIVE To systematically characterize the chemical constituents of Qikui Granules.METHODS Aqueous extract of Qikui Granules was analyzed by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS/MS)in both positive and negative ion modes.Aiding by PeakView software,both targeted and non-targeted approaches were employed to systematically screen and identify the chemical constituents from the obtained UPLC-QTOF-MS/MS raw data.Each screened compound was finally locked by the simultaneous occurrence of its corresponding quasimolecular ions.They were tentatively characterized or assigned according to its quasimolecular ions and diagnostic product ions,and then were confirmed by comparison with authentic standard,related database and literatures.RESULTS Totally 89 compounds were screened,and 71 of them were identified,including 45 flavonoids,14 saponins,4 anthraquinones,3 stilbene glycosides,2 polysaccharides and 3 organic acids.The results indicated that conversion reactions including hydroxylation,glucuronidation,acetylation,methylation,malonylation,as well as glycosidation(by the addition of one hexose,pentose,or tetrose),were involved in the formation of chemical constituents of Qikui Granules.CONCLUSION UPLC-QTOF-MS/MS based targeted and non-targeted compound screening approaches can clearly characterized the major constituents in Qikui Granules,providing a foundation for further researches on Qikui Granules such as quality control,preparation optimization and pharmacodynamics.