论文总字数：20593字

摘 要

非天然手性氨基酸及其衍生物在生物药品开发、化工合成、催化工业等方面具有十分广泛的应用，同时在化妆品、生化试剂领域也有开发的潜能。HPLC分析方法具有便捷、迅速、精确、精密度好的优点，如今普遍应用于手性氨基酸对映体的分离检测中。

这次毕业设计重点就在使用高效液相的检测方法来分析研究手性的对羟基苯甘氨酸(简称D-HPG)、N-氨甲酰基D-对羟基苯甘氨酸（简称N-HPG）以及对羟基苯海因（简称HPH），并对HPLC分析条件进行筛选优化，包括流动相配比，检测波长，流动相流速，柱温的选择等，使得未知混合样品有较好的分析和分离结果，并且最终确定了最理想的高效液相实验检测方案用以测定转化液中待测样品含量。同时对HPLC法进行考察，包括方法精密度考察，样品线性范围考察。将实验计算得到的关于样品的浓度与色谱峰高之间的关系来做对未知样品进行分析测定，从而得到了待测品的成分以及含量。

关键词：HPLC检测分析 D-对羟基苯甘氨酸 条件优化

HPLC analysis of chiral non-natural amino acids and their derivatives

Abstract

Non-natural chiral amino acids and their derivatives are widely used in drug development, chemical synthesis, catalytic industry and other aspects. Meanwhile, they are also widely used in the field of cosmetics and biochemical reagents.HPLC analysis is simple, fast, accurate, sensitive and reproducible. It has been used as an accurate and efficient method for amino acid detection.

In this paper, D-,hydroxyphenylglycine and its intermediates and hydroxyphenylhydantoin were analyzed by HPLC, and the HPLC analysis conditions were screened and optimized.The flow phase matching ratio, the detection wavelength, the flow velocity of the phase,including the selection of the column temperature, and so on, in order to make the main peak separated from the impurity peaks such as the raw material and the intermediate .Thus the best HPLC analysis program was established to determine the content of the samples to be tested in the conversion solution. Meanwhile, the HPLC method was investigated, including the precision of the method and the linear range of the samples.The content of D- hydroxyphenylglycine and its inrmediates and raw materials were calculated according to analysis of the linear relationship between the composition of unknown sample and the peak height given by laboratory.

Key Words: HPLC detect; D-,hydroxyphenylglycine ;condition optimization

目录

摘 要....................................................................I

ABSTRACT..................................................................II

第一章文献综述............................................................1

1.1 手性氨基酸概况........................................................1

1.1.1 手性非天然氨基酸及其衍生物简介................................1

1.1.2 手性非天然氨基酸及其衍生物的生理活性..........................1

1.2 手性非天然氨基酸及其衍生物的来源......................................2

1.3 手性氨基酸的应用与发展前景............................................2

1.4 手性氨基酸检测方法....................................................2

1.4.1 酶法检测........................................................2

1.4.2 毛细管电泳检测..................................................3

1.4.3 薄层色谱检测....................................................3

1.4.4 气相层析色谱检测................................................3

1.4.5 高效液相法检测.................................................4

1.5 课题意义 ............................................................7

第二章 实验材料与方法 .................................................. 8

2.1 实验材料..............................................................8

2.1.1 实验主要仪器....................................................8

2.1.2 实验主要试剂....................................................8

2.2. 实验方法.............................................................8

2.2.1 实验基本思路....................................................9

2.2.2 实验初始条件....................................................9

第三章 实验内容与过程 ..................................................10

3.1 系统适用性实验.......................................................10

3.2 色谱条件优化.........................................................10

3.2.1 流动相体积比 ..................................................10

3.2.2 最适pH.... ....................................................11

3.2.3 流动相流速.....................................................11

3.2.4 检测波长.......................................................11

3.3 实验方案的验证.......................................................12

3.3.1 精密度考察 ....................................................12

3.3.2 线性关系考察...................................................12

3.3.3 分离效果考察...................................................12

第四章 实验总结与分析...................................................13

4.1 最佳实验因素选择...................................................13

4.1.1 最初条件选择 ..................................................13

4.1.2 流动相配比的选择 ..............................................15

4.1.3 最适pH的选择 .................................................18

4.1.4 流动相流速的选择...............................................21

4.1.5 检测波长的选择.................................................24

4.2 实验方法验证.......................................................28

4.2.1 测定方法精密度...............................................28

4.2.2 线性关系考察.................................................29

4.2.3 分离效果考察.................................................31

4.3 实验展望...........................................................33

参考文献.................................................................34

致谢......................................................................36

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