鲨鱼硫酸软骨素二糖组成分析及其4S/6S比值的讨论

Disaccharide Composition of Shark Chondroitin Sulfate and Discussion on the Ratio of 4S/6S

目的研究鲨鱼来源硫酸软骨素的二糖组成,并对鲨鱼硫酸软骨素(shark-CS)4S/6S(N-乙酰半乳糖胺上4-硫酸化基团/N-乙酰半乳糖胺上6-硫酸化基团)比值进行探讨。方法通过酶解液相色谱法及文献比较对鲨鱼来源的硫酸软骨素二糖组成进行分析。结果鲨鱼硫酸软骨素主要由6-硫酸化软骨素二糖(ΔDi-6S)、4-硫酸化软骨素二糖(ΔDi-4S)、2,6-硫酸化软骨素二糖(ΔDi-2,6diS)以及无硫酸化的软骨素二糖(ΔDi-0S)组成,其百分含量分别为28%~65%、10%~44%、9%~25%、0.5%~23%。Shark-CS 4S/6S比值不超过0.8。结论提出Shark-CS 4S/6S比值的正确计算方法,解决了数据相互矛盾的问题。指出USP43中关于shark-CSΔDi-6S峰面积百分比大于ΔDi-4S峰面积百分比的不合理规定,为USP43修订提供参考与依据。

OBJECTIVE Research disaccharide composition of shark chondroitin sulfate and discussion on the ratio of shark-CS 4S/6S(4s/6s ratio as the ratio between the sulfated groups located in position 4 and 6 on N-acetyl-galactosamine).METHODS The chemical structure of shark chondroitin sulfate was analyzed by E-HPLC and comparison with the data in literature.RESULTS Shark-CS mainly containedΔDi-6S(28%-65%),ΔDi-4S(10%-44%),ΔDi-2,6diS(9%-25%),ΔDi-0S(0.5%-23%).A 4S/6S ratio no more than 0.8 in the case of shark-CS.CONCLUSION An accurate calculation method of shark-CS 4S/6S ratio is proposed,thus addressing the problem of conflicting data of previous reports.It was unreasonable that the requirement of the area percentage of theΔDi-6S peak is greater than that of theΔDi-4S peak in USP43.It has provided a reference for revising the USP43.