Yoshida K, Miyauchi S, Kikuchi H, Tawada A, Tokuyasu K
Tokyo Research Institute of Seikagaku Kogyo, Japan.
Anal Biochem. 1989 Mar;177(2):327-32. doi: 10.1016/0003-2697(89)90061-4.
A rapid and simple analytical method for unsaturated disaccharide isomers formed by enzymatic digestion from hyaluronic acid, chondroitin sulfate, dermatan sulfate, heparan sulfate, and heparin by high-performance liquid chromatography using an amine-bound silica column with a linear gradient of sodium dihydrogen phosphate was developed. The analyses were performed on isomers of two groups belonging to the chondroitin sulfate family and the heparin sulfate family. In both families, disaccharide isomers eluted in the order non-, mono-, di-, and trisulfated disaccharides by elevating salt concentrations. The method was applied to the analysis of constituent disaccharides of representative sulfated glycosaminoglycans, which proved that most constituents could be quantified separately. This method is advantageous in that enzymatic digests can be applied directly on a column without any pretreatment and good resolution of several disaccharides can be obtained by one chromatography.
开发了一种快速简便的分析方法，用于通过高效液相色谱法，使用胺基键合硅胶柱和磷酸二氢钠线性梯度，分析由透明质酸、硫酸软骨素、硫酸皮肤素、硫酸乙酰肝素和肝素经酶消化形成的不饱和二糖异构体。分析针对硫酸软骨素家族和硫酸乙酰肝素家族的两组异构体进行。在这两个家族中，二糖异构体通过提高盐浓度按非硫酸化、单硫酸化、二硫酸化和三硫酸化二糖的顺序洗脱。该方法应用于代表性硫酸化糖胺聚糖组成二糖的分析，结果表明大多数成分可以单独定量。该方法的优势在于酶消化物无需任何预处理即可直接进样到柱上，并且一次色谱分析就能获得几种二糖的良好分离度。
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