基于UHPLC-Q-Orbitrap-MS技术分析肝豆扶木汤中化学成分

Analysis of chemical constituents in Gandou Fumu Decoction by UHPLC-Q-Orbitrap-MS technology

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法(UHPLC-Q-Orbitrap-MS)对肝豆扶木汤(GDFMD)中化学成分进行快速鉴定和归属。方法:选择Thermo Fisher Vanquish UHPLC仪器,Aglient SB C_(18)色谱柱(2.1 mm×150 mm,1.8μm),流动相为0.1%甲酸水(A)和乙腈(B),洗脱梯度:0—2 min,3%B;2—7 min,3%~15%B;7—15 min,15%B;15—20 min,15%~40%B;20—22 min,40%~100%B;22—24 min,100%B;流速:0.4 mL/min;柱温:50℃;进样量:2μL。通过正、负离子模式采集质谱数据,根据精确相对分子质量、二级质谱裂解碎片等信息,结合对照品及相关文献信息比对,对GDFMD中化学成分进行鉴定,并对这些化合物的药材来源进行归属。结果:共鉴别了82个化合物,包括26个酚类、25个生物碱、18个皂苷、5个倍半萜、4个游离酚和4个有机酸;其中归属制何首乌8个、枸杞子24个、白芍15个、柴胡4个、三七粉14个、土茯苓16个、郁金6个。结论:采用UHPLC-Q-Orbitrap-MS可实现对GDFMD中化学成分的快速、准确鉴定,基本涵盖了组方中各药材的成分信息,可为完善该复方的质量评价体系和阐明其药效机制奠定基础。

Objective:To rapidly identify and attribute the chemical constituents in Gandou Fumu Decoction(GDFMD)by ultra-high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry(UHPLC-Q-Orbitrap-MS).Methods:Thermo Fisher Vanquish UHPLC instrument was selected,Aglient SB C_(18) column(2.1 mm×150 mm,1.8μm),mobile phase was 0.1%formic acid water(A)and acetonitrile(B),elution gradient was 0-2 min,3%B;2-7 min,3%-15%B;7-15 min,15%B;15-20 min,15%-40%B;20-22 min,40%-100%B;22-24 min,100%B;flow rate:0.4 mL/min;column temperature:50℃;injection volume:2μL.The mass spectrometry data was collected in positive and negative ion modes,and the chemical constituents in GDFMD were identified based on the information of accurate relative molecular mass,fragmentation fragments of secondary mass spectrometry,and the comparison of reference substances and related literature information.Results:A total of 82 chemical constituents were speculated and identified,including 26 phenolics,25 alkaloids,18 saponins,5 sesquiterpenoids,4 free phenolics and 4 organic acids.Among these identified chemical constituents,there were 8 from Polygoni Multiflori Radix Praeparata,24 from Lycium barbarum,15 from Paeoniae Radix Alba,4 from Bupleuri Radix,14 from Panax notoginseng(powder),16 from Smilacis Glabrae Rhizoma and 6 from Curcuma Radix.Conclusion:The use of UHPLC-QOrbitrap-MS can realize the rapid and accurate identification of the chemical constituent in GDFMD,which basically covers the constituent information of each medicinal material in the formula,and which can lay the foundation for improving the quality evaluation system of the compound and elucidating its pharmacological mechanism.