色谱

色谱

• 研究论文 • 上一篇    下一篇

手性衍生-高效液相色谱法拆分和定量测定茶氨酸对映体

李银花,刘仲华,黄建安

  

  1. Institute of Natural Products, Hunan Agricultural University, Changsha 410128, China

  • 收稿日期:2007-01-25 修回日期:2007-04-02 出版日期:2007-09-30 发布日期:1985-03-25
  • 通讯作者: 刘仲华

Separation and Quantification of Theanine Enantiomers Using High Performance Liquid Chromatography Coupled with Chiral Derivatization

LI Yinhua, LIU Zhonghua, HUANG Jian’an

  

  1. Institute of Natural Products, Hunan Agricultural University, Changsha 410128, China

  • Received:2007-01-25 Revised:2007-04-02 Online:2007-09-30 Published:1985-03-25

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摘要：

建立了以1-氟-2,4-二硝基苯基-5-L-丙氨酰胺（FDAA）为手性衍生试剂、高效液相色谱拆分茶氨酸对映体的方法。采用的色谱条件为：Kromasil C18色谱柱;三乙胺-磷酸缓冲液和乙腈为流动相,梯度洗脱，流速1.0 mL/min;检测波长为340 nm;柱温为35 ℃。L-茶氨酸的进样量在1.732×10-3～2.077 μg范围内峰面积与进样量之间的线性关系良好，加标回收率为97.3%～102.0%,检测限为4.973×10-4 μg,定量限为1.223×10-3 μg。D-茶氨酸的进样量在1.696×10-3～2.044 μg范围内峰面积与进样量之间的线性关系良好,加标回收率为97.2%～103.2%,检测限为5.871×10-4 μg,定量限为1.236×10-3 μg。该方法灵敏度高,分析过程中不发生消旋化。

关键词: 1-氟-2,4-二硝基苯基-5-L-丙氨酰胺（FDAA）, 茶氨酸对映体 , 高效液相色谱, 手性衍生

Abstract:

L-Theanine is a unique non-protein amino acid in tea, and shows many physiological functions. To directly extract L-theanine from tea is a lengthy and very costly process. Although the chemical synthesis of theanines is simple, the product is unfortunately a mixture of DL-enantiomers. A chiral derivatization method was developed for the separation and quantification of theanine enantiomers by reversed-phase high performance liquid chromatography （RP-HPLC）. 1-Fluoro-2,4-dinitrophenyl-5-L-alanine amide （FDAA）was used as the chiral reagent. This method showed good linearity for both L-theanine (ranging from 1.732×10-3 to 2.077 μg) and D-theanine (ranging from 1.696×10-3 to 2.044 μg). The recoveries were in the range of 97.3%-102.0% for L-theanine and 97.2%-103.2% for D-theanine. This method also showed excellent limit of detection (~5×10-4 μg) and limit of quantification (~1×10-3 μg) for both L-theanine and D-theanine. The results demonstrated that this method is precise, accurate and can be used for the determination of theanine enantiomers.

Key words: 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide （FDAA）, chiral derivatization, D-theanine enantiomers
, high performance liquid chromatography (HPLC)