Kitagawa H, Kinoshita A, Sugahara K
Department of Biochemistry, Kobe Pharmaceutical University, Japan.
Anal Biochem. 1995 Nov 20;232(1):114-21. doi: 10.1006/abio.1995.9952.
A series of disaccharides derived from chondroitin sulfate and heparin/heparan sulfate were derivatized at their reducing ends with the fluorophore 2-aminoacridone. The resulting labeled compounds derived from chondroitin sulfate or heparin/heparan sulfate were well separated and could be quantified by capillary electrophoresis and HPLC with a fluorescence detector. This method was successfully applied to the analysis of the disaccharide composition of sulfated tetra- and hexasaccharides derived from chondroitin sulfate and heparin/heparan sulfate. Remarkably, the saturated and the corresponding unsaturated chondrodisaccharides, the structures of which differ only in their nonreducing terminal uronic acid residues, were also well separated from each other by capillary electrophoresis. This facilitated disaccharide composition analysis of saturated chondrooligosaccharides prepared by testicular hyaluronidase digestion as well as unsaturated chondro- and heparin/heparan sulfate-oligosaccharides prepared by digestion with chondroitinases and heparitinases. The developed method employing the derivatization by 2-aminoacridone in conjunction with capillary electrophoresis or HPLC with a fluorescence detector allows a simple, sensitive, and quantitative disaccharide composition analysis of the glycosaminoglycan-derived oligosaccharides.
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-氨基吖啶进行衍生化。所得的源自硫酸软骨素或肝素/硫酸乙酰肝素的标记化合物得到了很好的分离，并且可以通过毛细管电泳和配备荧光检测器的高效液相色谱法进行定量。该方法成功应用于分析源自硫酸软骨素和肝素/硫酸乙酰肝素的硫酸化四糖和六糖的二糖组成。值得注意的是，饱和和相应的不饱和软骨二糖，其结构仅在非还原末端糖醛酸残基上有所不同，通过毛细管电泳也能很好地相互分离。这有助于对通过睾丸透明质酸酶消化制备的饱和软骨寡糖以及通过软骨素酶和肝素酶消化制备的不饱和软骨和肝素/硫酸乙酰肝素寡糖进行二糖组成分析。所开发的采用2-氨基吖啶衍生化结合毛细管电泳或配备荧光检测器的高效液相色谱法的方法，能够对糖胺聚糖衍生的寡糖进行简单、灵敏且定量的二糖组成分析。
