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

作　　者：王亮 江雪飞[1,2] 吴崇玉 熊志强 孙静怡 王明珠 孙化鹏 丛汉卿 乔飞[3] Wang Liang;Jiang Xuefei;Wu Chongyu;Xiong Zhiqiang;Sun Jingyi;Wang Mingzhu;Sun Huapeng;Cong Hanqing;Qiao Fei(Sanya Nanfan Research Institute of Hainan University,Sanya,572025;School of Tropical Agriculture and Forestry,Hainan University,Danzhou,571737;Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China(Ministry of Agriculture and Rural Affairs),Tropical Crop Germplasm Research Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101)

机构地区：[1]海南大学三亚南繁研究院,三亚572025 [2]海南大学热带农林学院,詹州571737 [3]中国热带农业科学院热带作物品种资源研究所,农业部华南作物基因资源与种质创制重点开放实验室,海口571101

出　　处：《分子植物育种》2024年第12期4033-4040,共8页Molecular Plant Breeding

基　　金：海南省自然科学基金面上项目(2019RC309);国家自然科学基金面上项目(NSFC:32070364)共同资助。

摘　　要：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.

关 键 词：海南粗榧 超高效液相色谱串联质谱 三尖杉碱 三尖杉类生物碱 三尖杉酸 

分 类 号：S791.53[农业科学—林木遗传育种]