用于分析糖蛋白和蛋白聚糖衍生寡糖的高效液相色谱分离模式比较

Comparison of separation modes of high-performance liquid chromatography for the analysis of glycoprotein- and proteoglycan-derived oligosaccharides.

作者信息

Davies M J, Hounsell E F

机构信息

Department of Biochemistry and Molecular Biology, University College London, UK.

出版信息

J Chromatogr A. 1996 Jan 12;720(1-2):227-33. doi: 10.1016/0021-9673(95)00221-9.

DOI:10.1016/0021-9673(95)00221-9

PMID:8601192

Abstract

Porous graphitised carbon (PGC) has been explored for high-performance liquid chromatography (HPLC) of mono- and di-saccharides released from proteoglycans and of fluorescently labelled oligosaccharide derivatives for high-sensitivity detection. Sulphated oligosaccharides show good retention and separation behaviour on PGC-HPLC, and compared to anion-exchange or reversed-phase ion-pair chromatography the chromatography is carried out in the absence of salt. Due to their poor retention on PGC-HPLC the analysis of single uronic acids has been optimised with high pH anion-exchange chromatography. Fluorescent labelled derivatives formed by reductive amination of neutral oligosaccharides with 2-aminobenzamide have been chromatographed on PGC-HPLC and by BioGel P4 gel filtration.

摘要

多孔石墨化碳（PGC）已被用于从蛋白聚糖释放的单糖和二糖以及荧光标记的寡糖衍生物的高效液相色谱（HPLC），以进行高灵敏度检测。硫酸化寡糖在PGC-HPLC上显示出良好的保留和分离行为，与阴离子交换或反相离子对色谱相比，该色谱在无盐条件下进行。由于单糖醛酸在PGC-HPLC上保留性差，因此已通过高pH阴离子交换色谱对其分析进行了优化。用2-氨基苯甲酰胺对中性寡糖进行还原胺化形成的荧光标记衍生物已在PGC-HPLC上进行色谱分析，并通过BioGel P4凝胶过滤进行分析。