UPLC同时测定海南粗榧中3种三尖杉生物碱类化合物  

Content Determination of Three Cephalotaxus Alkaloids in Cephalotaxus hainanensis by UPLC

作　　者：孙化鹏 王荣香[1] 丛汉卿[1] 乔飞[1] SUN Hua-peng;WANG Rong-xiang;CONG Han-qing;QIAO Fei(Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China,Ministry of Agriculture,Tropical Crops Genetic Resources Institute,Chinese Academy of Tropical Agricultural Science,Danzhou 571737,China)

机构地区：[1]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

出　　处：《中国实验方剂学杂志》2018年第19期47-51,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中央级公益性科研院所基本科研业务费专项(1630032017083,1630032017076);国家自然科学基金面上项目(31570326)

摘　　要：目的：海南粗榧中主要活性成分为以三尖杉碱为母核的三尖杉酯类生物碱,该研究利用超高压液相色谱（UPLC）建立一种同时测定三尖杉碱、三尖杉酯碱、高三尖杉酯碱3种化合物含量的检测方法,分析海南粗榧不同组织样品中3种生物碱的代谢分布。方法：选用Agilent 1290Ⅱ型超高压液相色谱仪,Agilent Infinitylab EC-C18色谱柱（2.1 mm×100 mm,2.7μm）,流动相甲醇-20 mmol·L碳酸铵水溶液,梯度洗脱,洗脱流速0.8 m L·min,检测波长291 nm,进样量5μL,柱温30℃。结果：在上述色谱条件下,色谱图基线平稳,各目标化合物峰型漂亮,分离度好。同时,对照品线性回归结果表明3种三尖杉生物碱含量与峰面积呈良好的线性关系,方法学考察实验结果表明仪器精密度、方法重复性及回收率均符合定量测定方法要求。结论：该方法具有较好的精密度、稳定性和重复性,且该方法检测时间短,12 min可以完成海南粗榧样品中3种三尖杉酯类生物碱的定量检测,操作简单快速、结果准确可靠,可以用于海南粗榧中三尖杉碱、三尖杉酯碱和高三尖杉酯碱的检测分析和质量控制,为深入研究海南粗榧中三尖杉酯类生物碱的代谢累积和时空分布等工作奠定方法学基础,同时也为海南粗榧的药用价值评价提供理论依据。Objective： The main active ingredients in Cephalotaxus hainanensis include cephalotaxine,harringtonine, homoharringtonine. This study aimed to establish a UPLC method for determination of cephalotaxine,harringtonine and homoharringtonine and analyze their metabolic accumulation and distribution in different tissues in C. hainanensis. Method： The method was performed on Agilent 1290 Ⅱ UPLC platform. The suitable conditions of UPLC were Agilent Infinitylab EC-C18 column（ 2. 1 mm × 100 mm,2. 7 μm, Agilent Infinitylab）,methanol and 20 mmol·L ammonium carbonate aqueous solution as mobile phase,gradient elution at 0. 8 m L·min,detective wavelength at 291 nm,injection volume of 5 μL,and column temperature at 30 ℃.Result： Under the above chromatographic conditions, the chromatogram baseline was stable,and each target compound had a good peak shape and resolution. The results showed a good linear relationship between content and peak area in all three C. hainanensis alkaloids. The methodological experiment showed that the instrument precision,method reproducibility and recovery rate all conform to the requirements of quantitative determination.Conclusion： This determination method has a good precision,stability and repeatability,with the advantages of shorter determination time, simpler operation and more accurate results. All of these three alkaloids could be detected in 12 min. It can be used to determine the contents of cephalotaxine, harringtonine and homoharringtonine in C. hainanensis,and establish a foundation for further study of metabolic accumulation in C.hainanensis.

关 键 词：超高效液相色谱 海南粗榧 三尖杉碱 三尖杉酯碱 高三尖杉酯碱 

分 类 号：R284.1[医药卫生—中药学] R289[医药卫生—中医学]