DNFB柱前衍生化RP-HPLC测定大黄种子氨基酸的含量  

Determination of Amino Acids in Rhubarb Seeds with DNFB Pre-column Derivatization by Reversed-phase High Performance Liquid Chromatography

作　　者：刘何春[1,2] 徐文华[1] 沈建伟[1] 宋文珠[1] 臧黎黎 周国英[1] 

机构地区：[1]中国科学院西北高原生物研究所,西宁810008 [2]中国科学院大学,北京100049 [3]电子科技大学,成都610041

出　　处：《天然产物研究与开发》2014年第7期1056-1061,共6页Natural Product Research and Development

基　　金：国家科技支撑计划项目(2011BA105B03)

摘　　要：采用柱前衍生RP-HPLC法测定大黄种子中氨基酸的含量。6mol／L盐酸水解得到氨基酸，以2，4-二硝基氟苯（DNFB）为柱前衍生化试剂，梯度洗脱。色谱柱为Gemlmi—NX C18（4．6mm×250mm，5μm）；流动相A相为0．1M乙酸钠水溶液，B相为乙腈-水（1：1），柱温37℃，检测波长360nm。结果表明，三种正品大黄种子中均含有17种氨基酸，总量在11．30％-19．26％。该方法灵敏、准确，有良好的重复性和稳定性。The aim of the study was to determine the contents of amino acids in rhubarb seeds with pre-column derivatization by reversed-phase high-performance liquid chromatography （RP-HPLC）. Amino acids extracted by the method of hydrolysis in 6 mol/L HCl were derivatized by 2,4-dinitrofluorobenzene （DNFB） and analyzed by RP-HPLC with gradient elution. The chromatographic separation was performed on a Gemimi-NX C18 （4.6 nun ×250 mm,5μm） column with 0.1 M aqueous sodium acetate solution （A） and acetonitrile-water （1： 1） （B） as mobile phases. The column tem- perature was maintained at 37℃, and the detection wavelength was set at 360 nm. The results showed that the three genuine rhubarb seed contained 17 kinds of amino acids, 11.30% to 19.26% of the total content. The experimental re- suits proved that the developed RP-HPLC method was sensitive,accurate and repeatable.

关 键 词：大黄种子 RP-HPLC 2 4-二硝基氟苯 氨基酸 

分 类 号：Q517[生物学—生物化学]