基于UPLC-Q-Orbitrap HRMS法比较不同炮制方法下山楂化学成分差异

Analysis of Chemical Constituents of Different Processing Methods of Crateagus by UPLC-Q-Orbitrap HRMS

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(UPLC-Q-Orbitrap HRMS)技术,鉴定山楂经3种不同炮制方法后的化学成分差异。方法:色谱采用Thermo Scientific Accucore TM C 18色谱柱(100 mm×3 mm,2.6μm),0.1%甲酸-水(A)和0.1%甲酸-乙腈(B)流动相,进行梯度洗脱;质谱采用ESI源,+/-离子模式扫描,扫描模式为Full MS/dd-MS2。利用Compound Discover 3.2软件分析数据,根据一级质谱质荷比、二级质谱碎片等信息,鉴定样品化学成分。结果:在炒山楂、焦山楂、山楂炭中共鉴定了49个化学成分,包括黄酮类、有机酸类、三萜酸类、苯丙素类、及其他类成分等。其中共有9种黄酮类成分、8种有机酸类、3种苯丙素类成分化学成分的含量发生了明显变化,表现为炒山楂>焦山楂>山楂炭。结论:利用UPLC-Q-Orbitrap HRMS法可全面、精确的测定炒山楂、焦山楂、山楂炭中的化学成分差异,进一步为山楂不同方法炮制品功效变化物质基础的确定提供了参考依据。

Objective:To analyze and identify the chemical constituents of 3 different processed methods of crateagus by an ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high resolution mass spectrometry(UPLC-Q-Orbirap HRMS)method.Methods:Compounds were isolated on Thermo Scientific Accucore TM C 18(100 mm×3 mm,2.6μm)column.The mobile phasewas 0.1%formic acid water and 0.1%formic acetonitrile with gradient elution.HRMS was performed using ESI,and scanned in positive and negative ion modes by means of Full MS/dd-MS2.Data analysis was performed using Compound Discover 3.2 software,and the composition identification was based on the information of accurate m/z of primary mass spectra and fragmentation of secondary mass spectra.Results:A total of 49 chemical components were identified in fried crateagus,charred crateagus and crateagus charcoal,including flavonoids,organic acids,triterpenoids,phenylpropanoids,and other components.A total of 9 flavonoids,8 organic acids,and 3 phenylpropanoids showed significant changes in chemical composition content,which showed that fried crateagus>parched crateagus>charred crateagus.Conclusion:The UPLC-Q-Orbitrap HRMS method can be used to comprehensively and accurately determine the differences of chemical components in fried crateagus,charred crateagus and crateagus charcoal,which further provides a reference basis for the determination of the material basis of the changes in the efficacy of different concoctions of crateagus.