基于UPLC-Q-TOF-MS/MS分析经典名方保元汤的物质基础

Analysis of Material Basis of Famous Classical Formula Baoyuantang Based on UPLC-Q-TOF-MS/MS

摘要:

目的 建立一种快速识别保元汤基准样品化学成分的检测方法，明确保元汤的物质基础。方法 基于超高效液相色谱-四极杆-飞行时间串联质谱法（UPLC-Q-TOF-MS/MS），系统地表征与鉴定保元汤水煎液中的化学成分。色谱条件为Waters ACQUITY UPLC HSS T3色谱柱（2.1 mm×100 mm，1.8μm），流动相0.1%甲酸水溶液（A）-0.1%甲酸乙腈溶液（B）梯度洗脱（0~3 min，2%~19%B；3~8 min，19%B；8~8.1 min，19%~22%B；8.1~14 min，22%~29%B；14~16 min，29%B；16~32 min，29%~45%B；32~32.1 min，45%~90%B；32.1~35 min，90%~95%B；35~36 min，95%~98%B；36~37 min，98%~2%B；37~40 min，2%B），基于电喷雾离子源（ESI），在正、负离子模式下检测，质量扫描范围m/z 50~1 500。通过UNIFI 1.9.4软件匹配、对照品比对、中医药百科全书（ETCM）数据库检索及文献报道等对保元汤水煎液中的化学成分进行系统解析。结果 在负离子模式下鉴定229个成分，正离子模式下鉴定181个成分，去重后共有322个成分，包括116个三萜皂苷类、66个黄酮类、19个有机酸类、6个姜酚类、6个姜醇类、5个姜酮类、10个氨基酸类、7个糖类、5个香豆素类和82个其他类化合物；其中人参归属83个成分，甘草归属141个成分，黄芪归属39个成分，肉桂归属35个成分，生姜归属38个成分。结论 该研究实现了保元汤水煎液中多类别成分的快速表征与鉴定，明确了保元汤的物质基础以三萜皂苷类、黄酮类、姜辣素类及有机酸类成分为主，并对化学成分进行药味归属，为后续的质量控制研究提供了参考。

Abstract:

Objective A rapid method for identification of chemical constituents in Baoyuantang reference sample was established in order to clarify the material basis of this formula.Method Based on ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） and self-established database information， the chemical components in Baoyuantang were systematically characterized and identified. The chromatography was performed on a Waters ACQUITY UPLC HSS T3 column（2.1 mm×100 mm， 1.8 μm） with mobile phase of 0.1% formic acid aqueous solution（A）-0.1% formic acid acetonitrile solution（B） for gradient elution（0-3 min， 2%-19%B； 3-8 min， 19%B； 8-8.1 min， 19%-22%B； 8.1-14 min， 22%-29%B； 14-16 min， 29%B； 16-32 min， 29%-45%B； 32-32.1 min， 45%-90%B； 32.1-35 min， 90%-95%B； 35-36 min， 95%-98%B； 36-37 min， 98%-2%B； 37-40 min， 2%B）. Based on electrospray ionization（ESI）， continuum data format was collected in both positive and negative ion modes with a scanning range of m/z 50-1 500. Chemical constituents in the decoction of Baoyuantang were systematically analyzed by UNIFI 1.9.4 software matching， control comparison， The Encyclopedia of Traditional Chinese Medicine（ETCM） database search and literature reports.Result A total of 229 components were identified under negative ion mode and 181 under positive ion mode， with a total of 322 components after removing duplicates， including 116 triterpene saponins， 66 flavonoids， 19 organic acids， 6 gingerphenols， 6 gingerols， 5 gingerones， 10 amino acids， 7 saccharides， 5 coumarins and 82 other components. Among them， 83， 141， 39， 35 and 38 components were attributed to Ginseng Radix et Rhizoma， Glycyrrhizae Radix et Rhizoma， Astragali Radix， Cinnamomi Cortex and Zingiberis Rhizoma Recens， respectively.Conclusion In this study， the rapid characterization and identification of multi-class components in Baoyuantang was realized， and it was confirmed that the material basis of this formula was mainly triterpenoid saponins， flavonoids， gingerols and organic acids， and the chemical composition was attributed and analyzed， which provided a reference for the subsequent quality control research.