基于UPLC-Q-Orbitrap HRMS和GC-MS技术的抗病毒颗粒化学成分分析

Chemical constituents of Antiviral Granules based on UPLC-Q-Orbitrap HRMS and GC-MS

目的:采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLC-Q-Orbitrap HRMS)技术和气相色谱-质谱联用(GC-MS)技术快速鉴定抗病毒颗粒的化学成分。方法:UPLC-Q-Orbitrap HRMS采用Thermo Scientific AccucoreTMC18色谱柱(3 mm×100 mm, 2.6μm),以甲醇-0.1%甲酸水为流动相进行梯度洗脱;正、负离子模式下采集数据。根据精确质荷比和二级碎片离子信息,结合对照品和文献数据对抗病毒颗粒的醇溶性成分进行快速鉴定。GC-MS选用Agilent HP-5毛细管色谱柱(30 m×250μm, 0.25μm),载气为氮气;质谱采用EMV模式:增益因子1.00。将色谱峰信息与NIST标准质谱库进行检索对比,对抗病毒颗粒的挥发性成分进行快速鉴定。结果:从抗病毒颗粒中共鉴别出了155个化合物,包括木脂素类8个、黄酮类15个、香豆素类9个、生物碱类9个、有机酸类31个、糖类6个、萜类38个、氨基酸类6个、核苷类7个、苯乙醇苷类11个和其他类化合物15个。结论:建立了抗病毒颗粒UPLC-Q-Exactive MS和GC-MS的分离鉴定方法,为阐释该复方的药效物质基础提供了依据。

OBJECTIVE The chemical constituents of Antiviral Granules were analyzed by ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry(UPLC-Q-Orbirap HRMS)and gas chromatography-mass spectrography(GC-MS)techniques.METHODS UPLC-Q-Orbirap HRMS analysis was performed on a Thermo Scientific AccucoreTMC18column(3 mm ×100 mm, 2.6 μm)ultra-performance liquid column.The mobile phase was methanol and 0.1% formic acid solution by gradient elution;MS data were collected in positive and negative ion modes.The alcohol-soluble components of Antiviral Granules were identified by combining the accurate mass charge ratio, secondary fragment ion information, reference substance, and literature data.GC-MS analysis was performed using AgilentHP-5(30 m×250 μm, 0.25 μm)capillary column with a carrier gas of nitrogen;mass spectrometry was performed using EMV mode: gain factor was 1.00.The volatile components of Antiviral Granules were rapidly identified by using the comparison of mass spectrometry information and NIST standard mass spectral library.RESULTS Totally 155 chemical constituents were identified in Antiviral Granules, including 8 lignans, 15 flavonoids, 9 coumarins, 9 alkaloids, 31 organic acids, 6 sugars, 38 terpenoids, 6 amino acids, 7 nucleosides, 11 phenylethanol glycosides and 15 other compounds.CONCLUSION The separation and identification methods of Antiviral Granules by UPLC-Q Exactive MS and GC-MS were established, which provided a foundation for further research on pharmacodynamic material basis of it.