| 用高效液相色谱定量分析分支链氨基酸 |
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| 引用本文： | 何晨光,马雷,徐庆阳,陈宁. 用高效液相色谱定量分析分支链氨基酸[J]. 生物技术通讯, 2009, 20(4): 556-558. DOI: 10.3969/j.issn.1009-0002.2009.04.030 |
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| 作者姓名： | 何晨光 马雷 徐庆阳 陈宁 |
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| 作者单位： | 1. 天津出入境检验检疫局,天津,300042
2. 天津科技大学,生物工程学院,天津,300457 |
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| 基金项目： | 天津市应用基础及前沿技术研究计划 |
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| 摘 要： | 目的：周2，4-二硝基氟苯（DNFB）对分支链氨基酸衍生后，采用优化的高效液相色谱（HPLC）对其进行定量分析。方法：色谱柱为AgilentZORBAXEclipsAAA（4．6mm×150mm，5-Micron），流动相为乙酸盐缓冲液（pH6．4）-乙腈，流速1．0mL／min，检测波长360nm。结果：用HPLC法测定分支链氨基酸的浓度为20-200mg／L时线性关系良好，3种分支链氨基酸的R。均在0．9997以上，平均回收率高，RSD≤0．56％（n=6）。结论：此方法快速、准确、重现性好，适合于对发酵液中分支链氨基酸的定量分析。
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| 关 键 词： | 分支链氨基酸 高效液相色谱 2，4-二硝基氟苯 |
Determination of the Content of Branched Chain Amino Acid by High Performance Liquid Chromatography |
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| HE Chen-Guang,MA Lei,XU Qing-Yang,CHEN Ning. Determination of the Content of Branched Chain Amino Acid by High Performance Liquid Chromatography[J]. Letters in Biotechnology, 2009, 20(4): 556-558. DOI: 10.3969/j.issn.1009-0002.2009.04.030 |
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| Authors: | HE Chen-Guang MA Lei XU Qing-Yang CHEN Ning |
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| Affiliation: | 1. Tianjin Entry-Exit Inspection and Quarantine Bureau, Tianjin 300042; 2. College of Bioteehnology, Tianjin University of Science and Technology, Tianjin 300457; China) |
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| Abstract: | Objective： An optimal method for determining branched chain amino acid（BCAA）） derived with 2,4-fluorodinitrobenzene（FDNB） using high performance liquid chromatography（HPLC） was reported. Methods： The HPLC analysis was carried out on Agilent ZORBAX Eclips AAA column（4.6 mm ×150mm, 5-Micron） with a mobile phase of the acetate buffer solution（pH6.4）-acetonitrile. The flow rate was 1.0 mL/min and UV detection was performed at 360 nm. Results： R2 of BCAA linear regression equation was beyond 0.999 7. The average recovery was better with RSD≤0.56%（n=6）. Conclusion： The results showed that this method is rapid, accurate and reproducible. It can be applied in quantitative determination of BCAA. |
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| Keywords: | branched chain amino acid high performance liquid chromatography 2,4-fluorodinitrobenzene |
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