基于UPLC-Q-TOF-MS和GC-MS的小金丸化学成分识别

Identification of chemical constituents in Xiaojin pills based on UPLC-Q-TOF-MS and GC-MS

目的:采用超高效液相色谱与串联四极杆飞行时间质谱(UPLC-Q-TOF-MS)和气相色谱-质谱联用(GC-MS)技术快速鉴定小金丸中的化学成分,构建较为全面的小金丸化学成分谱,为小金丸药效物质基础研究奠定基础。方法:UPLC-Q-TOF-MS分析采用ACQUITY UPLC BEH C_(18)(Waters,2.1 mm×100 mm,1.7μm)超高效液相柱,流动相为乙腈-0.1%甲酸水,梯度洗脱;飞行时间质谱采用正、负离子模式扫描,质谱数据以MSE模式采集。UPLC-Q-TOF-MS分析依据离子峰的保留时间、精确分子质量和多级质谱碎片,与文献报道数据和数据库对比,对小金丸的甲醇提取部分化学成分进行在线快速鉴定。GC-MS分析采用Agilent 19091S-433 HP-5MS 5%Phenyl Methyl Silox(Agilent,30 m×250μm×0.25μm)毛细管色谱柱,载气为氮气;质谱采用EMV模式:增益因子1.00。GC-MS分析通过增强型NIST4.L标准质谱库进行检索,对小金丸中挥发性成分进行了快速鉴定。结果:从小金丸中共鉴定化学成分118种,主要包括生物碱类24种、有机酸12种、甾体类9种、单萜类5种、倍半萜类16种、二萜类14种、三萜类7种和苯丙素类成分5种,另外还有个别酯类、糖类、氨基酸类和杂环类化合物等。结论:UPLC-Q-TOF-MS和GC-MS方法能快捷、准确、较全面地鉴定小金丸中的化学成分,为小金丸药效物质基础研究提供科学依据,并为中药复方的成分鉴定提供了一种简单快捷的分析鉴定方法。

OBJECTIVE To identify the chemical constituents and establish a comprehensive chemical composition electropherogram of Xiaojin pills by UPLC-Q-TOF-MS combined with GC-MS techniques.METHODS UPLC-Q-TOF-MS analysis was performed on an ACQUITY UPLC BEH C18(Waters,2.1 mm×100 mm,1.7μm)ultra-performance liquid column with a mobile phase of acetonitrile-0.1%formic acid water and gradient elution.Time-of-flight mass spectra were scanned in positive and negative ion mode.and the mass spectra data were collected in MSE mode.UPLC-Q-TOF-MS analysis was based on the retention time of the ion peak,the precise molecular weight and the multilevel mass spectrometry fragments.Compared with the data and database reported in the literature,the chemical composition of the methanol extraction part of Xiaojin pills was quickly identified.GC-MS analysis was performed using Agilent 19091 S-433 HP-5 MS 5%Phenyl Methyl Silox(Agilent,30 m×250μm×0.25μm)capillary column with a carrier gas of nitrogen;mass spectrometry was performed using EMV mode:gain factor was 1.00.GC-MS analysis was performed using the enhanced NIST4.L standard mass spectral library for rapid identification of volatile components in Xiaojin pills.RESULTS A total of 118 chemical constituents were identified in Xiaojin pills,including 24 kinds of alkaloids,12 kinds of organic acids,9 kinds of steroids,5 kinds of monoterpenes,16 kinds of sesquiterpenoids,14 kinds of diterpenes,7 kinds of triterpenes and 5 kinds of phenylpropanoids,as well as some esters,carbohydrates,amino acids and heterocyclic compounds.CONCLUSION UPLC-Q-TOF-MS and GC-MS methods can quickly,accurately,and comprehensively identify the chemical constituents in Xiaojin pills,providing a scientific basis for the study of therapeutic material basis,and it provides a simple and quick method for analysis and identification of the composition of traditional Chinese medicine compound.