DOLPA/AOT-异辛烷反胶束萃取-UPLC法测定倒提壶中4种生物碱的含量  

Determination of Four Alkaloids in Cynoglossum Amabile Stapf et Drumm by Ultrahigh Performance Liquid Chromatography with DOLPA/AOT-Isooctane Reverse Micelles Extraction

作　　者：樊轻亚[1] 王万好 代春美[3] FAN Qing-ya;WANG Wan-hao;DAI Chun-mei(Vocational and Technical College School of Medicinin of Xinyang,Xinyang464000,China;Henan Pharmaceutical Co.,Ltd.,Xinyang464000,China;China Academy of Chinese Medical Sciences,Beijing100700,China)

机构地区：[1]信阳职业技术学院,河南信阳464000 [2]河南同源制药有限公司,河南信阳464000 [3]中国中医科学院,北京100700

出　　处：《分析测试学报》2018年第12期1445-1450,共6页Journal of Instrumental Analysis

基　　金：河南省科技厅科技攻关项目(172102310315);信阳职业技术学院青年骨干教师培训计划

摘　　要：建立了二(2-乙基己基)磷酸酯(DOLPA)/二(2-乙基己基)丁二酸酯磺酸钠(AOT)-异辛烷反胶束萃取技术结合超高效液相色谱同时测定倒提壶中天芥菜品、天芥菜碱、毛果天芥菜碱及天芥菜定含量的方法。采用单因素试验考察了表面活性剂的种类与浓度、水相p H值、盐的种类与浓度、萃取相比、萃取及反萃取时间等因素对生物碱萃取率的影响。确定最佳萃取条件为:DOLPA/AOT的总浓度为0. 6 mol/L,水相p H值为4. 0,KCl浓度为0. 2 mol/L,萃取相比4∶1,萃取和反萃取时间分别为10、30 min。在优化条件下,以乙腈-0. 2%三乙胺为流动相进行超高效液相色谱法(UPLC)梯度洗脱,可在16 min内完成色谱分析,4种生物碱色谱峰的分离度良好;天芥菜品、天芥菜碱、毛果天芥菜碱及天芥菜定4种生物碱的质量浓度与峰面积呈良好线性关系,检出限分别为0. 030、0. 050、0. 040、0. 020 mg/L,回收率分别为98. 2%～103%、97. 9%～102%、98. 1%～101%、97. 8%～104%,精密度、重复性及加标回收率的RSD均不大于2. 3%。该法简单快捷、灵敏准确,满足倒提壶中4种生物碱的检测要求。A method of ultrahigh performance liquid chromatography(UPLC)with DOLPAAOT-isooctane reverse micelles was developed for the determination of heliosupine,heliotrine,lasiocarpine and heliotridine in Cynoglossum amabile Stapf et Drumm.Experimental conditions affecting the reverse micelles such as types and concentration of surface-active agent,pH value,salt species and concentration,ratio of sample and reverse micelles,extraction time and back extraction time were optimized.The optimal conditions were as followsDOLPA AOT concentration0.6 molL,pH value4.0,KCl concentration0.2 molL,ratio of sample and reverse micelles4∶1,extraction time10 min,back extraction time30 min.Under the optimal conditions,heliosupine,heliotrine,lasiocarpine and heliotridine were separated,and determined by UPLC with acetonitrile-0.2%triethylamine as mobile phase by gradient elution.Four ingredients were separated in 16 min.There existed good linear relationships for the analytes between peak areas and concentrations,with detection limits of 0.030,0.050,0.040 and 0.020 mgL,respectively.The recoveries were in the ranges of 98.2%-103%,97.9%-102%,98.1%-101%and 97.8%-104%,respectively.The RSDs for recovery,precision and reproducibility were not more than 2.3%.The method is convenient,sensitive and accurate,and is suitable for the determination and confirmation of 4 alkaloid in Cynoglossum amabile Stapf et Drumm.

关 键 词：倒提壶 生物碱 反胶束 超高效液相色谱法(UPLC) 

分 类 号：O657.72[理学—分析化学] TQ460.72[理学—化学]