UPLC-MS测定海南粗榧中3种三尖杉类生物碱和2种三尖杉酸

Determination of 3 Cephalotaxus Alkaloids and 2 Harringtonic Acids in Cephalotaxus hainanensis by UPLC-MS

为探究海南粗榧(Cephalotaxus hainanensis)中抗癌三尖杉酯碱、高三尖杉酯碱,以及其直接前体三尖杉碱、三尖杉酸和高三尖杉酸的含量和分布,本研究采用超声波甲醇提取海南粗榧样品中的生物碱及其前体,建立了超高效液相色谱串联质谱(UPLC-MS)的检测方法。色谱条件:色谱柱为Agilent InfinityLab EC-C18(2.1 mm×100 mm,2.7μm),流动相为0.1%v/v甲酸水溶液和乙腈,梯度洗脱,检测波长291 nm,柱温30℃,进样量2μL,流速0.3mL/min。质谱条件:质谱仪以Full MS/AIF模式运行,扫描碰撞能量35eV,喷雾电压3.5 kV,离子传输管温度275℃,辅助气加热温度350℃,鞘气流速35 arb,辅助气流速15 arb,吹扫气流速5 arb,辅助气、鞘气和吹扫气为高纯氮气(N_(2))。正离子模式检测3种生物碱,负离子模式检测2种酯碱的三尖杉酸。新建立的UPLC-MS检测方法在16 min内可以很好地分离和检测各种目标化合物,操作简单,结果准确可靠,可为海南粗榧中三尖杉碱的酯碱衍生物三尖杉酯碱、高三尖杉酯碱及其前体的定性定量分析提供参考。此外,不同组织中三尖杉酯碱、高三尖杉酯碱及其相应前体三尖杉酸的含量与生长年限呈正相关。

The contents and distributions of anti-cancer drugs in Cephalotaxus hainanensis,including harringtonine,homoharringtonine and their direct precursors of cephalotaxine,harringtonic acid and homoharringtonic acid were explored in this study.Ultrasonic bath and methanol were used to extract alkaloids and their precursors from Cephalotaxus hainanensis sample,and a UPLC-MS method was established for the detection.Chromatographic conditions:the chromatographic column Agilent Infinity Lab EC-C18(2.1 mm×100 mm,2.7μm)was gradient eluted with a mobile phase of 0.1%v/v formic acid in water and acetonitrile,detection wavelength of 291 nm,column temperature of 30 ℃,injection volume of 2μL,and flow rate of 0.3 mL/min.Mass spectrometric conditions:the mass spectrometer was operated in Full MS/AIF mode with a scan collision energy of 35 eV,spray voltage 3.5 kV,the temperature of that ion transmission tube 275℃,auxiliary gas heating temperature 350℃,sheath gas flow rate 35 arb,auxiliary gas flow rate 15 arb,sweep gas flow rate 5 arb;the auxiliary gas,sheath gas and sweep gas are high purity nitrogen(N_(2)).Three alkaloids were detected in positive mode,and two harringtonine acids were detected in negative mode.The newly established UPLC-MS detection method can separate and detect various target compounds within 16 min characterized with simple operation,precision and reliability,which can provide a reference for qualitative and quantitative analysis of harringtonine,homoharringtonine and their precursors in Cephalotaxus hainanensis.In addition,the contents of harringtoninevhomoharringtonine and their corresponding precursor harringtonine acids in different tissues are positive correlated with growing years.