NATURAL PRODUCT RESEARCH AND DEVELOPMENT ›› 2014, Vol. 26 ›› Issue (7): 1056-1061.

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

LIU He-chun1,2, XU Wen-hua1, SHEN Jian-wei1, SONG Wen-zhu1, ZANG Li-li3, ZHOU Guo-ying1*   

  1. 1 Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining 810008 ,China; 2 University of Chinese Academy  of Sciences, Beijing 100049 ,China;  3 University of Electronic Science and Technology of China,Chengdu 610041, China
  • Online:2014-07-31 Published:2014-11-09

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Abstract: 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 mm × 250 mm,5 μm) column with 0.1 M aqueous sodium acetate solution (A) and acetonitrile-water (1:1) (B) as mobile phases.The column temperature was maintained at 37 °C,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 results proved that the developed RP-HPLC method was sensitive,accurate and repeatable.

Key words: Rhubarb seeds, reversed-phase high performance liquid chromatography, 2,4-dinitrofluorobenzene, amino acids

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