食品科学 ›› 2010, Vol. 31 ›› Issue (16): 213-216.doi: 10.7506/spkx1002-6630-201016046

• 分析检测 • 上一篇    下一篇

柱前衍生反相高效液相色谱法测定柞蚕蛹蜕中氨基酸

杨 彬,阮长春,刘 志,孙光芝*   

  1. 吉林农业大学中药材学院
  • 收稿日期:2009-12-24 修回日期:2010-04-23 出版日期:2010-08-25 发布日期:2010-12-29
  • 通讯作者: 孙光芝* E-mail:bio-control@126.com

Determination of Amino Acids in the Pupal Exuviae of of Antheraea pernyi by Precolumn Derivatization Coupled with Reversed-Phase High Performance Liquid Chromatography

YANG Bin,RUAN Chang-chun,LIU Zhi,SUN Guang-zhi*   

  1. College of Chinese Medicinal Materia, Jilin Agricultural University, Changchun 130118, China
  • Received:2009-12-24 Revised:2010-04-23 Online:2010-08-25 Published:2010-12-29
  • Contact: SUN Guang-zhi* E-mail:bio-control@126.com

摘要:

建立一种反相高效液相色谱同时测定柞蚕蛹蜕中17 种氨基酸的分析方法。以正亮氨酸为内标物,异硫氰酸苯酯(PITC)作为Venusil-AA 氨基酸分析专用柱柱前衍生试剂,采用二元梯度洗脱,波长254nm 处检测柞蚕蛹蜕中氨基酸含量。17 种氨基酸在0.5~0.0125μmol 的范围内,具有良好的线性关系,加样回收率在96.2%~101.9%。柞蚕蛹蜕中氨基酸含量为21.9%,其中精氨酸含量最高。人体必需氨基酸占总氨基酸的33.62%。柱前衍生反相高效液相色谱法简单、高效,可以用于柞蚕蛹蜕氨基酸含量测定。

关键词: 高效液相色谱法, 柱前衍生, 异硫氰酸苯酯, 氨基酸, 正亮氨酸, 柞蚕蛹蜕

Abstract:

A reversed-phase high performance liquid chromatographic (RP-HPLC) method has been presented for the simultaneous determination of 17 amino acids in the pupal exuviae of Antheraea pernyi. Norleucine was used as internal standard for quantification using the internal standard method. For amino acid analysis, a Venusil-AA amino acid column was used, and phenylisothiocyanate (PITC) was the precolumn derivatization agent. Elutions were carried out in a binary gradient mode. The detection wavelength was set at 254 nm. The calibration curves of 17 amino acids all exhibited good linearity over the range from 0.5 to 0.0125μmol. Mean recoveries for 17 amino acids were between 96.2% and 101.9%. The total amino acid content of the pupal exuviae of Antheraea pernyi was determined to be approximately 21.9%, and the total essential amino acid content represented 33.62% of the total amino acids, and the content of arginine was the highest. This method is simple and efficient and provides a good method for the determination of amino acids in he pupal exuviae of Antheraea pernyi.

Key words: high performance liquid chromatography, pre-column derivatization, phenylisothiocyanate, amino acid, norleucine, the pupal exuviae of Antheraea pernyi

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