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1.0mL/min;检测波长为340nm;柱温为35℃。L-茶氨酸的进样量在1.732×10-3～2.077μg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4.973×10-4μg,定量限为1.223×10-3μg。D-茶氨酸的进样量在1.696×10-3～2.044μg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5.871×10-4μg,定量限为1.236×10-3μg。该方法灵敏度高,分析过程中不发生消旋化。
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. 077μg） and D-theanine （ranging from 1.595 × 10^-3 to 2. 044 μg）. The recoveries were in the range of 97.3% - 102.0% for L-theanine and 97.2% - 105.2% 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.
出处
《色谱》
CAS
CSCD
北大核心
2007年第5期719-722,共4页
Chinese Journal of Chromatography
