基于UPLC-Q-TOF MS及GC-MS技术分析脑血栓专利方全成分

Chemical Composition Analysis of the Patented Naoxueshuan Formula Based on UPLC-Q-TOF MS and GC-MS

  • 摘要: 本研究采用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF MS)法和气相色谱-质谱(GC-MS)法分析脑血栓专利方的化学成分。对于非挥发性成分,采用Waters BEH C18色谱柱(100 mm×2.1 mm,1.7 μm),以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,电喷雾离子源正、负离子模式采集数据;对于挥发性成分,采用DB-5MS毛细管柱(30 m×0.25 mm,0.25 μm)在程序升温下分离,EI离子源电离,质量扫描范围m/z 50~550。根据精确质荷比、二级质谱信息、NIST标准谱库检索及相关文献鉴定脑血栓专利方成分,并对化合物进行来源归属。结果表明,在脑血栓专利方中共鉴定出61种非挥发性成分以及29种挥发性成分,其中包括14种黄酮类、13种萜类、9种苷类、5种有机酸类、17种烃类、3种醇类、1种醛类、1种酯类、3种酚类、2种萜烯类以及12种其他类型的化合物。本研究可为揭示脑血栓专利方的药效物质基础提供数据支持。

     

    Abstract: The Patented Naoxueshuan formula is an empirical formula for treating cerebral thrombosis, which has been proven to have significant and stable therapeutic effects through long-term clinical application. The Naoxueshuan formula has rich chemical components and diverse structures. In this study, chemical compositions of the Patent Naoxueshuan formula were analyzed by using ultra-performance liquid chromatography-time of flight mass spectrometry (UPLC-Q-TOF MS) and gas chromatography-mass spectrometry (GC-MS). For UPLC-Q-TOF MS analysis, the decoction of Naoxueshuan formula was separated using a Waters BEH C18 column (100 mm×2.1 mm, 1.7 μm). The mobile phase was comprised of 0.1% formic acid solution and acetonitrile with the flow rate of 0.3 mL/min. MS data was collected under positive and negative ion modes. For GC-MS analysis, the Agilent DB-5MS capillary column (30 m×0.25 mm, 0.25 μm) was employed to analyze the volatile components of the decoction of Naoxueshuan formula. The GC-MS analysis was performed under programmed heating conditions with electron ionization (EI) as the ion source and a scanning mass range of m/z 50-550. The main chromatographic peaks in both liquid and gas chromatograms were identified and analyzed according to the secondary mass spectrum information, NIST standard spectrum library and literatures. A total of 61 non-volatile components are identified in the Naoxueshuan decoction, including 14 flavonoids, 13 terpenoids, 9 glycosides, 5 organic acids, and 10 other types of compounds. A total of 29 volatile components are identified in the Naoxueshuan decoction, including 17 hydrocarbons, 3 alcohols, 1 aldehydes, 1 esters, 3 phenols, 2 terpenes and 2 other types of compounds. Among them, the organic acids are mainly from Salvia miltiorrhiza Bunge, the flavonoids are mainly from Pueraria lobata, Astragalus membranaceus (Fisch.) Bunge and Carthamus tinctorius L., the terpenoids are mainly from Salvia miltiorrhiza Bunge and Cornus officinalis Sieb. et Zucc. The volatile components are mainly from Ligusticum sinense, Fructus Amomi, Alpinia katsumadai Hayata, and Semen Alpiniae Katsumadai. This study provides an effective and comprehensive method for qualitative analysis of the chemical components in the Patented Naoxueshuan formula, and also provides data support for clarifying effective substances and establishing a quality evaluation system.

     

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