食品中氨基酸检测方法的研究

文档编号:SW028  文档字数:9791,页数:22

摘 要：本文档研究了一种用高效液相色谱分析技术（HPLC）测定食品中（主要是奶粉中）氨基酸的含量。本方法在衍生过程中使用了OPA和FMOC两种试剂，其中，OPA作为一级衍生剂，FMOC作为二级氨基酸衍生剂。通过对流动相、流速、衍生剂、衍生时间、检测器等的研究，确定氨基酸的检测方法。实验采用色谱柱Hypersil ODS氨基酸分析柱，长度20cm，内径2.1mm，柱温40.0℃，流速：0.45 ml／min，流动相：醋酸钠与三乙胺的水溶液调节PH至7.310.05作为流动相A,醋酸钠与三乙胺的水溶液调节PH至7.200.05，再与甲醇和乙腈混合作为流动相B，通过检测器参数切换程序达到最佳检测波长。该方法RSD在l.83% ~4．93%之间，重现性较好．样品加标回收率在89.30% ~ll5.70% 之间，线性范围10~80 pmol／L，最小检出限为0.25tanol／L，相关系数均在0.9980以上，此法具有操作简便易行、准确度高、重现性好等优点.

5 ●●关键词：高效液相色谱；氨基酸；食品

The research on detection a method for amino acid in food

Abstracts: my thesis aims to find a special method to detect the NH4 content in food by HPLC. Two reagents--- OPA and FMOC, are used in the spin-off process, the former of which is used as a kind of level of derivative and the latter of which is used as a kind of one derivative of NH4 based on protein secondary structures. This research includes using a Hypersil ODS column (20cm×2.1mm)and the column temperature is 40℃。Then the flow rate is 0.45ml/min. As for the mobile phase, there are totally two. The mobile phase A is sodium acetate—triethylamine(PH﹦7.20±0.05) and the mobile phase B is methanol—acetonitrile mixed with the mobile phase A . At last, use the  DPSS detector to turn the UV detection into the best condition .When RSD is in the range of1.83％～4.93％，this method has better reproducibility. The recovery rate of the sample is in the range of 89.3％～115.7％, the linear relationship is appeared in the range of 10～80pmol/L ,the minimal limit of assay is in the range of 0.25tanol/L and the correlation coefficients are all more than 0.9980. In conclusion, this method has an advantage of great convenience, precision and reproducibility.
Key words: HPLC; amino acid; food

目  录
中文摘要 …………………………………………………………………………………Ⅰ
英文摘要 …………………………………………………………………………………Ⅱ
目录 ………………………………………………………………………………………Ⅲ
1. 绪论 ……………………………………………………………………………………1
 1.1前言…………………………………………………………………………………1
1.2氨基酸简介…………………………………………………………………………1
1.3食品中氨基酸分析…………………………………………………………………1
1.4高效液相色谱法……………………………………………………………………1
  1.5高效液相色谱法原理、特点、应用范围…………………………………………2
2. 实验部分 ………………………………………………………………………………4
 2.1 仪器和试剂  ……………………………………………………………………4
 2.2 实验试剂配制……………………………………………………………………4
 2.3 实验方法…………………………………………………………………………5
3. 结果与讨论 ……………………………………………………………………………7
 3.1 标准曲线的制作 ……………………………………………………………… 7
 3.2 梯度的选择 ………………………………………………………………………9
 3.3 流动相PH值选择…………………………………………………………………9
 3.4 流速的选择 ………………………………………………………………………9
 3.5 衍生时间的选择 …………………………………………………………………9
 3.6 衍生剂的选择 ……………………………………………………………………9
 3.7 检测器的选择 ……………………………………………………………………10
 3.8 样品的定性与定量 ………………………………………………………………10
 3.9 三乙胺的影响 ……………………………………………………………………10
 3.10  不同色谱条件下分离情况考察 ………………………………………………11
 3.11 回归方程、线性范围和检测限结果 ………………………………………… 11
 3.12 样品氨基酸种类和含量测定结果 …………………………………………… 12
 3.13 重现性和回收率实验结果 …………………………………………………… 15
4. 总结与展望 …………………………………………………………………………… 15
致谢………………………………………………………………………………………… 17
参考文献…………………………………………………………………………………… 18

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