Kodama C, Ototani N, Isemura M, Yosizawa Z
J Biochem. 1984 Oct;96(4):1283-7. doi: 10.1093/oxfordjournals.jbchem.a134947.
A sensitive method was developed for the separation and quantitation of four unsaturated disaccharides (delta Di-0S, delta Di-4S, delta Di-6S, and delta Di-diS) by high performance liquid chromatography. The unsaturated disaccharides were coupled with a fluorescent compound, 2-aminopyridine. Complete separation of the resulting pyridylamino derivatives was achieved on a column of muBondapak-C18 with 8 mM KH2PO4-Na2HPO4 (pH 6.0)/methanol (30/l, by volume) as a mobile phase. There was a linear relationship between the fluorescence emission (peak height), and the amount of each authentic disaccharide used for the coupling reaction. This method was applied to analyze commercially available chondroitin sulfates A and C, dermatan sulfate, and urinary glycosaminoglycans obtained from patients with mucopolysaccharidosis after digestion with chondroitinases. The data indicated that the present method is useful for the separation and quantitation of nmol-pmol levels of the unsaturated disaccharides produced from chondroitin sulfate isomers by chondroitinases and can be used for their structural characterization.
开发了一种灵敏的方法，用于通过高效液相色谱法分离和定量四种不饱和二糖（δDi-0S、δDi-4S、δDi-6S和δDi-diS）。不饱和二糖与荧光化合物2-氨基吡啶偶联。以8 mM KH2PO4-Na2HPO4（pH 6.0）/甲醇（体积比30/1）作为流动相，在μBondapak-C18柱上实现了所得吡啶基氨基衍生物的完全分离。荧光发射（峰高）与用于偶联反应的每种纯二糖的量之间存在线性关系。该方法应用于分析市售硫酸软骨素A和C、硫酸皮肤素以及从黏多糖贮积症患者中提取的经软骨素酶消化后的尿糖胺聚糖。数据表明，本方法可用于分离和定量软骨素酶从硫酸软骨素异构体产生的纳摩尔-皮摩尔水平的不饱和二糖，并可用于其结构表征。
