痛泻要方水煎液化学成分的UPLC-LTQ-Orbitrap-MS分析

Chemical Constituent Analysis of Tongxie Yaofang Decoction by UPLC-LTQ-Orbitrap-MS

目的:采用超高效液相色谱-线性离子阱-静电场轨道阱高分辨质谱法(UPLC-LTQ-Orbitrap-MS)快速鉴定痛泻要方水煎液中的化学成分。方法:色谱条件为ACQUITY UPLC BEH C_(18)色谱柱(2.1 mm×100 mm,1.7μm),流动相0.1%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~4 min,5%~15%B;4~10 min,15%~25%B;10~15 min,25%~60%B;15~20 min,60%~90%B;20~25 min,90%~100%B;25~27 min,100%B;27~30 min,100%~5%B;30~32 min,5%B),流速0.3 m L·min^(-1),进样量3μL,柱温35℃。质谱条件为电喷雾离子源(ESI),扫描范围m/z 100~1 250,正、负离子模式下分别采集MS、MS/MS数据。结合对照品、数据库和文献信息,运用Trace Finder 4.1和Xcalibur 2.1软件对痛泻要方水煎液进行化学成分鉴定。结果:在正、负离子模式下,从痛泻要方水煎液中共鉴定出90个化合物,主要为黄酮类、香豆素类、单萜糖苷类、色原酮类、内酯类。通过对所有化合物的药材来源进行归属,发现其中9个化合物来源于白术,21个化合物来源于白芍,24个化合物来源于陈皮,29个化合物来源于防风,另有7个化合物来源于至少2味药材。结论:该方法能快速、有效、全面地识别痛泻要方水煎液中的化学成分,明确了痛泻要方化学组成。鉴定出的化合物涵盖了4味组方药材的主要活性成分且丰度较高,说明痛泻要方提取方法和药材配比具有一定科学性,可为该方物质基础和质量评价研究提供参考。

Objective:To rapidly identify the chemical constituents in Tongxie Yaofang decoction by ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-LTQ-Orbitrap-MS).Method:Chromatographic conditions were ACQUITY UPLC BEH C_(18) column(2.1 mm×100 mm,1.7μm),mobile phase of 0.1%formic acid aqueous solution(A)-acetonitrile(B)for gradient elution(0-4 min,5%-15%B;4-10 min,15%-25%B;10-15 min,25%-60%B;15-20 min,60%-90%B;20-25 min,90%-100%B;25-27 min,100%B;27-30 min,100%-5%B;30-32 min,5%B),flow rate of 0.3 mL·min-1,column temperature at 35℃and injection volume of 3μL.UPLC-LTQ-Orbitrap-MS was equipped with an electrospray ionization(ESI),the MS and MS/MS data were collected in positive and negative ion modes,and detection range was m/z 100-1250.Combining the reference substance,chemical databases and related literature information,TraceFinder 4.1 and Xcalibur 2.1 were used to identify the chemical constituents of Tongxie Yaofang decoction.Result:A total of 90 compounds,mainly including flavonoids,coumarins,monoterpene glycosides,chromones and lactones,were identified from Tongxie Yaofang decoction.By attributing the sources of Chinese medicines for all identified compounds,9 of them were found to be derived from Atractylodis Macrocephalae Rhizoma,21 from Paeoniae Radix Alba,24 from Citri Reticulatae Pericarpium,29 from Saposhnikoviae Radix,and 7 from at least two Chinese medicines.Conclusion:The method can effectively,quickly and comprehensively identify the chemical components of Tongxie Yaofang decoction,and clarify the chemical composition.These identified compounds cover the main active ingredients of the four herbs with high abundance,which indicates that the extraction method and the ratio of the medicinal materials of Tongxie Yaofang are scientific,and can provide a reference for the research on the material basis and quality evaluation of this famous classical formula.