基于超高效液相色谱-串联质谱检测三黄膏主要成分  

Detection of main components of Sanhuang Ointment based on UPLC-MS/MS

作　　者：王天明 易剑锋[1] 王波[1] 潘海邦[1] WANG Tianming;YI Jianfeng;WANG Bo;PAN Haibang

机构地区：[1]甘肃中医药大学,甘肃兰州730000

出　　处：《中医临床研究》2024年第6期8-15,共8页Clinical Journal Of Chinese Medicine

基　　金：国家自然基金项目(81860850);甘肃省教育厅——优秀研究生“创新之星”项目(2021CXZX-734,2021CXZX-736)。

摘　　要：目的:采用超高效液相色谱-串联质谱(Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry,UPLC-MS/MS)对三黄膏的化学成分进行分析和鉴定。方法:取三黄膏50 mg加入甲醇内标提取液,涡旋后取上清液,用微孔滤膜过滤后准备检测。液相条件:Agilent SB-C18 1.8μm,2.1 mm×100 mm色谱柱,以超纯水(加入0.1%的甲酸)和乙腈(加入0.1%的甲酸)为流动相,梯度洗脱,流速0.36 mL/min,柱温40℃,进样量2μL。质谱条件:电喷雾离子源温度500℃;离子喷雾电压5 500 V(正离子模式)/–4 500 V(负离子模式);离子源气体I(GSI)、气体II(GSII)和气帘气分别设置为50、60和25 psi,碰撞诱导电离参数设置为高。三重四级杆扫描使用多反应检测(Multiple Reaction Monitoring,MRM)模式,并将碰撞气体(氮气)设置为中等。通过进一步的去簇电压(Declustering Potential,DP)和碰撞能(Collision Energy,CE)优化,完成了各个MRM离子对的DP和CE。根据每个时期内洗脱的代谢物,在每个时期监测一组特定的MRM离子对。采用电喷雾离子源,正负离子模式下采集数据。结果:共鉴定和推测出30个化合物,包括11种生物碱类、11种黄酮类、3种萜类、5种酚酸类。结论:通过UPLC-MS/MS鉴定的30种化合物与黄芩、黄连、黄柏主要化合物进行对比后发现,经过加工处理后的三黄膏保留了黄芩、黄连、黄柏的主要化学成分。为今后三黄膏的物质基础研究奠定了基础。Objective:To analyze and identify the chemical components of Sanhuang Ointment(三黄膏)by ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Methods:A total of 50 mg of Sanhuang Ointment was taken and added into methanol internal standard extract.After swirling,supernatant was taken and filtered by microporous filter membrane for detection.Liquid phase conditions:Agilent SB-C181.8μm,2.1 mm×100 mm column with ultra-pure water(with 0.1%formic acid)and acetonitrile(with 0.1%formic acid)as mobile phases,gradient elution was performed,flow rate was 0.36 m L/min,column temperature was 40℃,injection volume was 2μL.Mass spectrometry conditions:electrospray ionization(ESI)temperature was 500℃.Ion spray voltage was 5500 V(positive ion mode)/–4500 V(negative ion mode).Ion source gas I(GSI),gas II(GSII)and curtain gas were set to 50,60 and 25 psi,respectively,and collision-induced ionization parameters were set to high.QQQ scanning was performed using MRM mode and the collision gas(nitrogen)was set to medium.Further declustering potential(DP)and collision energy(CE)optimization was accomplished for DP and CE of individual MRM ion pairs.A specific set of MRM ion pairs was monitored in each period depending on the metabolites eluted within each period.Data was collected in positive and negative ion mode by electrospray ion source.Results:A total of 30 compounds were identified and postulated,including 11 alkaloids,11 flavonoids,3 terpenoids,and 5 phenolic acids.Conclusion:Comparison of the 30 compounds identified by UPLC-MS/MS with the main compounds of Huangqin(Scutellaria baicalensis),Huanglian(Radix Scutellariae),and Huangbo(Radix Phellodendron)reveals that the processed Sanhuang Ointment retains the main chemical components of Huangqin,Huanglian,and Huangbo.It lays the foundation for future research on the material basis of Sanhuang Ointment.

关 键 词：三黄膏 超高效液相色谱-串联质谱 化学成分谱 

分 类 号：R289.5[医药卫生—方剂学]