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2-氨基蒽醌柱前衍生化糖类物质的分析
The Analysis of Saccharides Derivatised with 2-Aminoanthraquinone
【作者】 逄鑫；
【导师】 王仲孚；
【作者基本信息】 西北大学 ， 细胞生物学， 2011， 硕士
【摘要】 高效液相色谱(HPLC)已经成为糖类物质分离分析的重要工具。由于糖类物质没有光学活性,因此需要通过衍生化的方法使之带上紫外或荧光集团才能实现在HPLC上分离检测。目前已经开发了很多种类的衍生化试剂,但是大多都存在一些不足如衍生化流程繁琐、不同的反应样品需要摸索不同的反应条件、产生副产物、质谱检测灵敏度不高等,所以寻找新的性能优良的衍生化试剂很有意义。本文首次选用了2-氨基蒽醌作为糖的标记试剂,建立了糖柱前衍生化的高效液相色谱(HPLC)以及电喷雾质谱(ESI-MS)分析方法,取得了较好的效果,表明该方法在糖类物质的分析方面有潜在的应用价值。本研究主要得到了以下的结果和结论。1.实验中探讨了各种衍生化条件对产率的影响,并对这些条件加以优化。优化后的衍生化方法条件温和,70℃反应60min即可完成,反应完全。2.将五种单糖标准进行柱前衍生化,考察了此衍生化方法的稳定性、线性关系和最低检测限(LOD)、回收率等指标,建立了较为理想的单糖分析模式,成功地测定了白芨BSPI-A组分的单糖组成。3.将2-氨基蒽醌用于寡糖混合物的分离,建立了一套寡糖混合物柱前衍生化的HPLC-ESI-MS组分分析方法。表明该方法在食品体系寡糖混合物分离分析方面有的应用前景。
【Abstract】 High Performance Liquid Chromatography(HPLC) had became an important tool in the analysis of carbohydrates due to its efficiency and high-speed. because of the sugers had no optical activity,they can not be well detected on the HPLC until we supplied a ultraviolet or fluorescence group for them through derivatization reaction.A great many of derivatization reagent had been introduced,but they had many shortages such as its complex derivatization procedures,its different reaction conditions for different samples,its by-products,its less sensitivity on MS. So it was worth to try new functional reagents which can meet the need of analyzing the sugers.In this paper,we established a new HPLC and ESI-MS analysis method of carbohydrates by 2-Aminoanthraquinone pre-column derivatization based on reductive amination,and the results were nice. The developed dervatizationi method had potential application outlook for the analysis of mixed oligosaccharides.1. Derivatization conditions which affected the reaction productivity were discussed and optimized. The reaction was very mild and can be completed under 70℃、60min.2. The five monosaccharide standers were pe-column derived using 2-Aminoanthraquinone. we studied the reaction indexs such as stability、linear relation and lowest detectable limit and so on.Then established desired analysis model for monosaccharides. We successfully analyzed the nonosaccharides composition of BSPI-A from Bletilla striata.3. The means to separate and analyze mixed oligosaccharides using HPL、ESI-MS、LC-ESI-MS was successfully established, which was based on a new reagent 2-Aminoanthraquinone. This method had potential application prospect for the analysis of complicated oligosaccharides of food system.