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| 2,4-二硝基氟苯衍生法测定游离氨基酸方法的优化 |
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张宏杰,周建军,李新生,吴三桥,李会宁
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陕西省资源生物重点实验室,陕西,汉中,723000
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| 摘要: |
| 以美国HP LC1100系列为基本实验设备,在流速为1.0ml/min, 紫外检测器波长为360nm,运行时间25min,进样量为10ul情况下,筛选了流动相梯度、2,4-二硝基氟苯用量、柱温,测定了回收率.结果表明:FDBN衍生法分析游离氨基酸,流动相Ⅰ为0.04N KH2PO4(pH=7.2±0.05,40%KOH调整)缓冲液;流动相Ⅱ为乙腈的水溶液,其浓度为55%,流动相Ⅰ的百分比梯度为86/0 min-88/2min-86/4min-70/10 min-30/20min-10/21 min-0/24.0min,1%FDBN用量为5-20ul,柱温40 ℃,样品AA浓度大于0.5nmol/ul,17种AA分离效果最佳.此方法AA回收率在100%～104%之间. |
| 关键词: 2,4-二硝基氟苯 氨基酸 测定方法 优化 |
| DOI： |
| 分类号:Q503 |
| 基金项目:陕西省教育厅资助项目 |
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| Optimization for Determining Free Amin Acid with FDBN Method |
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| Abstract: |
| An optimal method for determining free amino acid derived with FDBN by using HPLC was reported here,The chromatography was run with 0.04M KH 2PO 4 and 55%of CH 3CN as mobilephase ,and an UV detector.It was found that the 17 amino acids could be separated with 100% 104% of reeovery when the column temperature was at 40℃ and the concentration of aa was higher than 0.5n mol·L 1 . |
| Key words: FDBN,Assay of amino acids,Optinization? |
