基于UPLC-Q-TOF-MS技术分析百合地黄汤的化学成分

Analysis of Chemical Constituents in Baihe Dihuangtang by UPLC-Q-TOF-MS

目的:采用超高效液相色谱-四级杆飞行时间串联质谱(UPLC-Q-TOF-MS)技术对百合地黄汤中的化学成分进行研究。方法:选择ACQUITY UPLC BEH C18色谱柱(2.1 mm×50 mm,1.7μm),以乙腈(A)-0.1%甲酸水溶液(B)为流动相进行梯度洗脱(0~3 min,5%~11%A;3~6 min,11%~12%A;6~8 min,12%A;8~12 min,12%~20%A;12~13 min,20%~26%A;13~15 min,26%~32%A;15~17 min,32%~42%A;17~19 min,42%~5%A;19~20 min,5%A),流速0.4 m L·min-1,柱温30℃,进样量5μL;采用电喷雾离子源,在正、负离子模式下采集数据。运用SCIEX OS 1.4软件,根据化合物的相对保留时间、准分子离子峰和碎片离子峰等信息,结合对照品和相关文献资料进行化学成分分析。结果:在百合地黄汤中共鉴定出49个化学成分,包括酚酸甘油酯类5个,苯乙醇苷类15个,环烯醚萜类11个,紫罗兰酮类8个,苯丙素类3个,核苷类2个,有机酸类1个,其他类4个;其中酚酸甘油酯类来源于百合,其他成分主要来源于地黄,4个化学成分(毛蕊花糖苷、异毛蕊花糖苷、阿魏酸、咖啡酸)经对照品比对鉴定。结论:该研究建立了能快速、准确分析百合地黄汤化学成分的检测方法,较全面地阐述了百合地黄汤的化学成分信息,为该方的质量控制、药效物质基础研究以及复方制剂开发奠定了基础。

Objective:To study the chemical constituents in Baihe Dihuangtang by ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS).Method:The separation was performed on an ACQUITY UPLC BEH C18 column(2.1 mm×50 mm,1.7μm)by a gradient elution of acetonitrile-0.1%formic acid solution.The flow rate was 0.4 m L·min-1 and the column temperature was 30℃,the injection volume was 5μL.Electrospray ionization was applied and the data were collected via positive and negative ion modes.By using SCIEX OS 1.4 software,the chemical constituents were analyzed based on the retention time,excimer ion peak and fragment ion peak of the compounds,as well as comparison with reference substances and literature data.Result:A total of 49 chemical constituents in Baihe Dihuangtang were identified,including 5 phenolic glycerides,15 phenylethanoid glycosides,11 iridoids,8 lonones,3 phenylpropanoids,2 nucleosides,1 organic acid,and 4 other compounds.Among them,phenolic glycerides belonged to Lilii Bulbus,and other components mainly belonged to Rehmanniae Radix.Four chemical constituents(acteoside,isoacteoside,ferulic acid and caffeic acid)were identified by comparison of reference substances.Conclusion:The established detection method can quickly and accurately analyze the chemical constituents of Baihe Dihuangtang.The information of chemical constituents in Baihe Dihuangtang is comprehensively expounded.The study establishes a foundation for the research of quality control,material foundation of efficacy and the development of compound preparations of Baihe Dihuangtang.