摘要
目的:建立用柱前衍生和高效毛细管电泳测定地龙中水解氨基酸含量的方法。方法:地龙药材经水煎煮后,先用盐酸水解处理地龙样品,再用Waters AccQ-Fluor衍生试剂对水解氨基酸进行柱前衍生,采用高效毛细管电泳方法测定地龙中氨基酸的含量。结果:在pH 2浓度0.25 mol.L-1的H3PO4-Na2HPO4电泳缓冲液,进样量为0.5 psi×4 s,分离电压20 kV电泳条件下,在16 min内14种常规氨基酸分离效果较好,氨基酸浓度15.625～250μmol.L-1线性关系良好,成功测定地龙水煎液中氨基酸的含量。结果显示地龙药材中Glu,Ile/Leu,Gly,Ala含量相对较高,His,Tyr含量较低,Asp未检测到。结论:采用高效毛细管电泳进行AQC衍生后地龙氨基酸的分析,操作简单、分析时间短,测定结果可靠,可适用于动物类中药材中氨基酸的快速检测。
Objective： To develop a method using pre-column derivation and high-performance capillary electrophoresis for determining amino acids content in pheretima. Method： Pheretima, a kind of Chinese medicinal material was firstly treated with water boiling, and then hydrolyzed by hydrochloride acid. The hydrolysate of amino acids in pheretima was derived by Waters AccQ-Fluor reagent. The amino acids in pheretima were identified and quantified with the method of high-performance capillary electrophoresis. Result： The optimized CZE conditions were as follows： a running buffer composed of 25 mmol·L-1 H3PO4-NaEHPQ （pH 2） , separation voltage at 20 kV, temperature at 25 centigrade, injection time for 4 sec with 0.5 psi pressure. 15 kinds of amino acids in pheretima were determined within 16 minutes. The separation effect of amino acids was perfect and the linear relationship was good in the range of 15. 625-250 μmol·L-1 amino acids concentration. The results showed that there was relative high content of Glu, Ile/Leu, Gly, Ala and low content of His, Tyr in pheretima of Chinese medicinal material. And Asp was not detected. Conclusion： The present established method of AQC derivation and high-performance capillary electrophoresis is simple, rapid and suitable for determination of amino acids in animal medicinal materials.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第2期117-120,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
浙江省教育厅科研项目(Y200908742)
关键词
地龙
氨基酸
含量
毛细管电泳
pheretima
amino acids
content
high performance capillary electrophoresis （HPCE）
