基于UHPLC-Q-OrbitrapHRMS技术的参桂胶囊中主要化学成分研究

Main chemical constituents of Shengui Capsules based on UHPLC-Q-OrbitrapHRMS

目的为更加全面、系统、快速地表征参桂胶囊有效化学成分谱,采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(UHPLC-Q-OrbitrapHRMS)技术对参桂胶囊中主要化学成分进行快速识别和归属分类。方法采用ACQUITY UPLC?BEH C18色谱柱(100 mm×2.1 mm,1.7μm),以乙腈(A)-0.1%甲酸水(B)为流动相进行梯度洗脱,以实现化合物的前期分离;然后通过Q-Orbitrap MS正、负离子同时监测,一级全扫描及自动触发二级质谱扫描的模式捕捉未知成分的精确相对分子质量及碎片离子信息,同时与对照品的保留时间和质谱数据信息进行匹配,并结合相关文献以及Chemical Book、Mass Bank等数据库最终实现对中药中复杂成分的快速准确识别。结果本研究共鉴定出40种化学成分,其中包括16种皂苷类、6种有机酸类、13种内酯类及5种其他类。结论本法可快速、准确地识别参桂胶囊中主要化学成分,同时为其进一步的药效物质基础、质量控制及临床合理应用等研究奠定了科学的前期基础。

Objective In order to study the chemical profile of Shengui Capsules, a novel ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry(UHPLC-Q-OrbitrapHRMS) was applied to establish a method to recognize and classify the ingredients accurately and rapidly. Methods ACQUITY UPLC? BEH C18 chromatographic column(100 mm × 2.1 mm, 1.7 μm) was employed and the separation was performed with the mobile phase consisting of acetonitrile-0.1% formic acid aqueous solution. The information of accurate mass and multistage fragment ions were obtained by the novel 'monitored simultaneously for positive and negative ions, Full MS scan and automatic trigger secondary mass spectrometry' mode of Q-Orbitrap MS technology. Then, the complex chemical constituents would be identified by comparing the relative retention time and the mass data of the reference substance, and consulting the reference literature or Mass Bank, Chemical Book network database at the same time. Results Forty chemical components were finally identified in this study, including the 16 tritepene saponins, 6 organic acids, 13 lactones, and 5 other classes. Conclusion The method established in this study could be used to rapidly, accurately, and systematically identify the main chemical constituents of Shengui Capsules. Moreover, this study will lay a scientific preliminary foundation for the further pharmacodynamic material basis, quality control improvement and the rational clinical use.