CAPILLARY ELECTROPHORESIS FOR THE ANALYSIS OF CHONDROITIN SULFATE-DERIVED AND DERMATAN SULFATE-DERIVED DISACCHARIDES
CAPILLARY ELECTROPHORESIS FOR THE ANALYSIS OF CHONDROITIN SULFATE-DERIVED AND DERMATAN SULFATE-DERIVED DISACCHARIDES
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DOI：
10.1016/0003-2697(91)90296-6
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发表时间：
1991-05-15
影响因子：
2.9
通讯作者：
LINHARDT, RJ
中科院分区：
文献类型：
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作者：
ALHAKIM, A;LINHARDT, RJ
高压毛细管区带电泳（CZE）已被用于分析的非，单，二，和三硫酸化的二糖硫酸软骨素，硫酸皮肤素，透明质酸的第一次。这些糖胺聚糖首先使用多糖裂解酶解聚。所得不饱和二糖产物可通过其在232 nm处的紫外吸光度来检测。通过CZE分析，获得了每种不饱和二糖的不同保留时间。使用单一的，简单的缓冲液系统，在70厘米的熔融石英毛细管两端的恒定电压的应用程序解决了在40分钟内的八组分的混合物。来自硫酸软骨素使用软骨素ABC裂解酶（EC 4.2.2.4）和混合物的不饱和二糖标准的二糖定量是可能的，只需要皮克量的样品。所检测的二糖的净电荷为-1至-4，主要根据净电荷进行解析，其次根据电荷分布进行解析。分析了两种均含有相同不饱和糖醛酸残基的未硫酸化二糖。一种来自具有N-乙酰半乳糖残基的软骨素，一种来自具有N-乙酰葡糖残基的透明质酸盐。尽管事实上，他们的区别仅在一个中心的手性，这些二糖的CZE解决。毛细管电泳法是一种快速、简便的方法，为糖胺聚糖酸性二糖组分的分析和分离提供了一种强有力的新工具。
High-voltage capillary zone electrophoresis (CZE) has been used for the first time in the analysis of non-, mono-, di-, and trisulfated disaccharides derived from chondroitin sulfate, dermatan sulfate, and hyaluronic acid. These glycosaminoglycans are first depolymerized using polysaccharide lyases. The resulting unsaturated disaccharide products can be detected by their ultraviolet absorbance at 232 nm. Different retention times were obtained for each unsaturated disaccharide analyzed by CZE. The application of a constant voltage across a 70-cm fused silica capillary using a single, simple buffer system resolved an eight-component mixture within 40 min. Quantitation of disaccharides derived from chondroitin sulfate using chondroitin ABC lyase (EC 4.2.2.4) and mixtures of unsaturated disaccharide standards was possible requiring only picogram quantities of sample. The disaccharides examined had a net charge of from −1 to −4 and were resolved primarily on the basis of net charge and secondarily on the basis of charge distribution. Two unsulfated disaccharides both containing the same unsaturated uronic acid residue were analyzed. One was from chondroitin having an N-acetylgalactosyl residue and one from hyaluronate having an N-acetylglucosyl residue. Despite the fact that they differed only by the chirality at one center, these disaccharides were resolved by CZE. CZE is a fast and simple method that represents a powerful new tool for analysis and separation of acidic disaccharide components of glycosaminoglycans.