UHPLC-Q-Orbitrap HRMS快速鉴定羊红膻药材的化学成分

Rapid Identification of Chemical Constituents of Pimpinella thellungiana by UHPLC-Q-Orbitrap HRMS

目的:应用超高液相色谱四级杆-静电场轨道肼高分辨质谱联用(UHPLC-Q-Orbitrap HRMS)技术,快速识别与鉴定羊红膻药材中主要化学成分。方法:采用Waters Acquity UPLC BEH C18色谱柱(2.1 mm×50 mm,1.7μm),流动相0.1%甲酸水溶液(A)-乙腈(B)进行梯度洗脱。质谱采用ESI源,负离子一级全扫描-数据依赖性二级质谱扫描(MS2)模式采集数据。通过高分辨质谱数据解析,并结合对照品数据和相关文献对羊红膻药材中的化学成分进行识别和鉴定。结果:通过精确相对分子质量和二级碎片离子分析,并结合对照品确认、质谱裂解规律和相关文献检索,共鉴定29个化合物,包括绿原酸类化合物19个,黄酮类7个,小分子有机酸3个,其中除木犀草素葡萄糖醛酸苷和芹菜素葡萄糖醛酸苷外,其余27个化合物为首次从该药材中发现。结论:建立的UHPLC-Q-Orbitrap HRMS可快速鉴定羊红膻药材中的化学成分,研究结果为其药效物质基础、质量控制以及临床应用等研究提供了科学的依据。

Objective:Ultra high performance liquid chromatography coupled with Q-Exactive Focus hybrid quadrupole and orbitrap high resolution mass spectrometer(UHPLC-Q-Orbitrap HRMS)was applied for identification of chemical constituents in Pimpinella thellungiana.Method:Chromatographic separation was performed on a WATERS BEH C18 column(2.1 mm×50 mm,1.7μm).The mobile phase consisted of 0.1%formic acid aq(A)and acetonitrile(B)with a gradient elution.Mass spectral analysis were performed on QExactive Focus hybrid quadrupole and orbitrap mass spectrometer.The mass spectrometer was connected to UHPLC instrument via an ESI interface.Samples were analyzed in negative ion mode by the full-scan-dd MS2(datadependent MS/MS)scanning mode.Then the constituents of P.thellungiana were identified by compared HRMS data with those of the standard compounds,MS cleavage mechanism and the related literatures.Result:Based on the characteristic mass data of accurate molecular weight and fragmentation ion information,29 chemical constituents were identified including 19 chlorogenic acids,7 flavonoids and 3 phenolic acid.Among them,the identified components except luteolin-7-O-β-D-glucuronopyranoside and apigenin-7-O-β-D-glucuronopyranoside were reported in P.thellungiana for the first time.Conclusion:The established UHPLC-Q-Orbitrap HRMS method can be used to identify the chemical constituents of P.thellungiana quickly and accurately,providing the scientific evidence for its quality evaluation and material basis research.