【作者】 孙燕；

【导师】 吕志华； 徐家敏；

【作者基本信息】 中国海洋大学 ， 生药学， 2004， 硕士

【摘要】 褐藻胶寡糖是由β-1，4-D-甘露糖醛酸（M）和α-1，4-L-古罗糖醛酸（G）两种单体通过1-4糖苷键连接而成的海洋酸性寡糖，具有抗病毒、增强免疫功能、抗肿瘤及抗炎等生物活性，与内源性糖链中的许多结构有相似之处，有着巨大的开发潜力。本论文首次利用高效毛细管电泳中的区带电泳，进行了褐藻胶寡糖的高效毛细管电泳分离分析方法研究。 酶降解的褐藻胶寡糖在非还原端碳4，5位形成双键与碳6位羧基中的羰基共轭，在235nm处具有特征紫外吸收。本论文采用235nm的紫外光直接进行检测，进行了酶降解褐藻胶寡糖的高效毛细管电泳分离分析方法的研究。优化了电泳条件，实现了高效分离。 酸降解的褐藻胶寡糖与酶解寡糖不同，在碳4，5位没有形成双键，没有紫外吸收特征，也没有荧光发光基团，对其进行分析检测比较困难。利用柱前衍生技术，与衍生试剂8-氨基芘．1，3，6-三磺酸钠（APTS）反应，引入荧光基团，采用毛细管电泳-激光诱导荧光检测方法，建立了标准褐藻胶寡糖的毛细管电泳-激光诱导荧光检测分离分析方法。优化了衍生条件，并考察了电泳缓冲液介质、浓度、pH值、工作电压及分离温度对寡糖-APTS衍生物分离的影响，实现了酸降解褐藻胶寡糖的高效分离，检测限达到了10^-17mol水平，确定了标准曲线及线性范围。同时应用该分析方法对褐藻胶的水解产物进行了分离分析，检测了聚甘露糖醛酸和聚古罗糖醛酸的酸降解产物，进行了褐藻胶全水解产物M／G比值的测定，都获得了良好的分离分析效果。 褐藻胶寡糖是酸性寡糖，其电泳条件和电泳行为与文献报道的中性糖有不同之处，本文根据电泳结果进行了讨论。海洋酸性寡糖其结构特点与内源性寡糖有很多相似点，所以酸性寡糖毛细管电泳分离分析方法的研究对糖缀合物的分析也有重要的意义。更多还原

【Abstract】 Alginate oligosaccharides is a kind of marine acidic oligosaccharides made of -1,4-D-mannuronic acid and -1,4-L- guluronic acid with good anti-virus, anti-tumor and anti-inflammation activities. In this paper, alginate oligosaccharides are separated and analyzed by high performance capillary electrophoresis.Alginate oligosaccharides degraded by the alginate lyase are separated by HPCE with UV 235nm detection. CE separation conditions have been investigated. The oligosaccharides are analyzed in conditions of pH 2.5, 100mmol/L phosphate buffer, 20kV, temperature 30 C. In this condition alginate oligosaccharides are successfully separated.Alginate oligosaccharides degraded by acid are derivatized with 8-aminopyrene-l,3,6-trisultonate (APTS) by reductive animation and their ARTS derivatives are separated and detected by capillary zone electrophoresis with laser-induced fluorescence. Opitimization studies have also been performed with respect to labeling and HPCE separation conditions of oligosaccharides. The detection sensitivity can reach 10~17mol, standard curve and linearity range are determined. The alginate oligosaccharides- APTS derivatives are separated in an acidic phosphate buffer (pH 2.5) and in the alkaline borate buffer (pH 10.02), respectively. Acid hydrolysis products of alginate labeled with APTS are successfully mapped and the M/G ratio of alginate is also determined by this method.There are some comparabilities in structure and the properties of electrification between marine acidic oligosaccharides and endogenous substances, so the study of acidic oligosaccharides by HPCE has important significance.更多还原