Murata K, Yokoyama Y
J Chromatogr. 1987 Apr 10;415(2):231-40. doi: 10.1016/s0378-4347(00)83215-6.
An improved high-performance liquid chromatographic (HPLC) method for unsaturated disaccharides prepared from hyaluronic acid and various chondroitin sulphate and dermatan sulphate isomers was developed, which involves an ion-exchange resin prepared from a sulphonated styrene-divinylbenzene copolymer. The retention times of the individual unsaturated disaccharides were unique and reproducible, the disaccharides appearing in the following order: unsaturated non-sulphated disaccharide derived from hyaluronic acid, then unsaturated 6-sulphated, non-sulphated and 4-sulphated disaccharides from chondroitin sulphate isomers. Unsaturated disulphated disaccharide G had a much shorter retention time than the unsaturated non-sulphated disaccharide derived from hyaluronic acid. The contents of these individual unsaturated disaccharides could be determined with similar sensitivities on the basis of their ultraviolet absorbance. Selective and unique retention times and good resolutions were found for various unsaturated disulphated and trisulphated disaccharides. The proposed method can be used to determine various chondroitin sulphate and dermatan sulphate isomers in addition to hyaluronic acid in amounts as small as 100 ng to 8 micrograms. The practicality of this method was verified by its application to the separation and determination of the different types of chondroitin sulphate and dermatan sulphate isomers derived from human arteries in the presence of appreciable amounts of hyaluronic acid.
开发了一种改进的高效液相色谱（HPLC）方法，用于分析由透明质酸以及各种硫酸软骨素和硫酸皮肤素异构体制备的不饱和二糖，该方法涉及一种由磺化苯乙烯-二乙烯基苯共聚物制备的离子交换树脂。各个不饱和二糖的保留时间是独特且可重现的，二糖按以下顺序出现：源自透明质酸的不饱和非硫酸化二糖，然后是来自硫酸软骨素异构体的不饱和6-硫酸化、非硫酸化和4-硫酸化二糖。不饱和二硫酸化二糖G的保留时间比源自透明质酸的不饱和非硫酸化二糖短得多。这些单个不饱和二糖的含量可以根据其紫外吸光度以相似的灵敏度进行测定。对于各种不饱和二硫酸化和三硫酸化二糖，发现了选择性和独特的保留时间以及良好的分离度。所提出的方法可用于测定低至100 ng至8 μg的透明质酸以及各种硫酸软骨素和硫酸皮肤素异构体。通过将该方法应用于在存在大量透明质酸的情况下分离和测定源自人动脉的不同类型的硫酸软骨素和硫酸皮肤素异构体，验证了该方法的实用性。
