Kinoshita A, Sugahara K
Department of Biochemistry, Kobe Pharmaceutical University, Kobe, Higashinada-ku, 658-8558, Japan.
Anal Biochem. 1999 May 1;269(2):367-78. doi: 10.1006/abio.1999.4027.
A series of disaccharides derived from chondroitin sulfate and heparin/heparan sulfate were derivatized at their reducing ends with a fluorophore 2-aminobenzamide to develop a sensitive microanalytical method for glycosaminoglycans. The resulting labeled compounds derived from chondroitin sulfate or heparin/heparan sulfate were well-separated and quantified by HPLC equipped with a fluorescence detector. The detection limit was a low picomole level. This method was applied to the analysis of the disaccharide composition of tetra- and hexasaccharides derived from chondroitin sulfate and heparin/heparan sulfate as well as these glycosaminoglycan polysaccharides. The method was also successfully applied to the exosequencing of chondrohexasaccharides, where the fluorophore-labeled oligosaccharides were degraded exolytically from the nonreducing ends using bacterial eliminases. The resultant labeled fragments were identified by HPLC.
一系列源自硫酸软骨素和肝素/硫酸乙酰肝素的二糖在其还原端用荧光团2-氨基苯甲酰胺进行衍生化，以开发一种用于糖胺聚糖的灵敏微分析方法。所得源自硫酸软骨素或肝素/硫酸乙酰肝素的标记化合物通过配备荧光检测器的高效液相色谱法进行了良好的分离和定量。检测限为低皮摩尔水平。该方法应用于分析源自硫酸软骨素和肝素/硫酸乙酰肝素的四糖和六糖以及这些糖胺聚糖多糖的二糖组成。该方法还成功应用于软骨六糖的外切测序，其中荧光团标记的寡糖使用细菌消除酶从非还原端进行外切降解。所得标记片段通过高效液相色谱法进行鉴定。
