加味独活寄生合剂及含药血清主要成分超高效液相色谱-四级杆-时间飞行质谱仪分析

Analysis of Main Components of Jiawei Duhuo Jisheng Mixture(加味独活寄生合剂)and Drug-Containing Serum by Ultra Performance Liquid Chromatography-Quadrupole-Time of Flight Mass Spectrometer

目的:通过超高效液相色谱-四级杆-时间飞行质谱仪(UPLC-Q-TOF)分析加味独活寄生合剂及含药血清,筛选出其主要物质成分及入血清成分。方法:采用Agilent Extend C18(2.1 mm×50 mm,1.8μm)色谱柱,流动相:乙腈-水(水中含0.1%甲酸,质谱级)按规定条件进行梯度洗脱,流速:0.4 mL/min,进样量:2μL,质谱检测条件:离子化方式为电喷雾离子化(ESI),准确质量数用ESI-L Low Concentration Tuning Mix(G1969-85000)校正,正离子分析模式,通过比较样品峰、保留时间等质谱信息,并结合提取离子流图及与对照品、相关文献数据对比进行化学成分确认。结果:(1)加味独活寄生合剂UPLC-Q-TOF分析确认了47个共有峰,通过分析比对主要包括亚麻酸甲酯、腺苷、邻苯二甲酸二异丁酯、金合欢基丙酮、十二酸甘油酯等47个化合物,主要来源为加味独活寄生合剂处方中的独活、桑寄生、杜仲、秦艽等,当前的条件分析尚未检测到细辛的药物成分。(2)加味独活寄生合剂含药血清UPLC-Q-TOF分析指认33个共有峰,通过分析比对主要包括邻苯二甲酸二异丁酯、金合欢基丙酮、十二酸甘油酯、邻苯二甲酸丁辛酯、桃叶珊瑚苷等33个化合物,主要来源为加味独活寄生合剂处方中的独活、桑寄生、杜仲、制天南星、黄芩、木瓜、威灵仙等。当前的条件分析加味独活寄生合剂含药血清中尚未检测到细辛、肉桂、秦艽、当归4味药物成分。结论:UPLC-Q-TOF指认了加味独活寄生合剂47个共有峰,含药血清33个共有峰,从整体上系统地反映加味独活寄生合剂的物质化学信息,初步确认了33个入血化学成分,以提示加味独活寄生合剂的药效物质基础,为药效实验提供了参考和依据。

Objective:To analyze Jiawei Duhuo Jisheng Mixture and drug-containing serum by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometer(UPLC-Q-TOF),and to screen out the main substance components and the components in serum.Methods:An Agilent extend C18(2.1 mm×50 mm,1.8μm)column was used.The mobile phase was acetonitrile water(containing 0.1%formic acid in water,mass spectrometry grade).The flow rate was 0.4 mL/min,and the injection volume was 2μL.Mass spectrometry detection conditions:ionization mode is electrospray ionization(ESI),accurate mass was calibrated with ESI-L Low Concentration Tuning Mix(G1969-85000),positive ion analysis mode.The chemical composition is confirmed by comparing the mass spectrum information such as sample peak and retention time,and combining the extracted ion current diagram and comparison with the reference substance and relevant literature data.Results:(1)The UPLC-Q-TOF analysis of Jiawei Duhuo Jisheng Mixture confirmed 47 common peaks.Through analysis and comparison,they mainly included methyl linolenate,adenosine,diisobutyl phthalate,farnesylacetone,glycerol dodecyl ester,etc.The main source were Duhuo(Angelicae pubescentis radix),Sangjisheng(Mistletoe),Duzhong(Eucommiae cortex)and Qinjiao(Gentianae macrophyllae radix)in the prescription of the Jiawei Duhuo Jisheng Mixture.The current condition analysis has not detected the ingredients of Xixin(Asarum).(2)The UPLC-Q-TOF analysis of drug-containing serum of Jiawei Duhuo Jisheng Mixture confirmed 33 common peaks.Through analysis and comparison,they mainly included diisobutyl phthalate,farnesylacetone,glycerol dodecyl ester,butyl octyl phthalate,aucubin,etc.The main source were Duhuo(Angelicae pubescentis radix),Sangjisheng(Mistletoe),Duzhong(Eucommiae cortex)and Zhitiannanxing(Arisaema),Huangqin(Scutellaria),Mugua(Chaenomeles),Weilingxian(Clematis)in the prescription of the Jiawei Duhuo Jisheng Mixture.The current conditions have not detected the four medicine components of Xixin(Asarum),Rougui(Cinnamomi Cortex),Qinjiao(Gentianae macrophyllae radix),Danggui(Chinese angelica)in the drug-containing serum of Jiawei Duhuo Jisheng Mixture.Conclusion:UPLC-QTOF has identified 47 common peaks of Jiawei Duhuo Jisheng Mixture and 33 common peaks for drug-containing serum,which systematically reflected the material chemical information of Jiawei Duhuo Jisheng Mixture and preliminarily confirmed 33 chemical components in blood,so as to indicate the pharmacodynamic material basis of Jiawei Duhuo Jisheng Mixture and provide reference and basis for efficacy experiment.