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目的制备并结构解析一种硫酸乙酰肝素十二糖。方法采用肝素酶Ⅱ将硫酸乙酰肝素(HS)降解得到寡糖混合物,经分子筛Bio-Gel P-10低压柱色谱分离得到大小均一的十二糖混合物,再经强阴离子交换高效液相色谱(SAX-HPLC)分离,得到电荷均一的十二糖纯品。该寡糖经肝素酶Ⅰ、Ⅱ、Ⅲ完全降解后进行二糖组成分析,并采用电喷雾碰撞诱导串联质谱(ES-CIDMSn)确定其结构序列。结果从HS寡糖混合物中分离得到了一种大小和电荷均一的十二糖纯品,它由ΔUA-GlcNAc(Ⅳ-A)与ΔUA-GlcNS(Ⅳ-S)组成,其相对比例为5∶1,经ES-CID-MSn分析,确定其结构序列为ΔUA-GlcNAc-GlcA-GlcNS-GlcA-GlcNAcGlcA-GlcNAc-GlcA-GlcNAc-GlcA-GlcNAc。结论本实验从HS中制备了一种十二糖,并对其结构进行解析,为该HS寡糖构效关系研究奠定了基础。
Objective To prepare and structurally elucidate a heparan sulfate heptadecane. Methods Heparinase (II) was used to degrade heparan sulfate (HS) to obtain a mixture of oligosaccharides. The mixture was separated by Bio-Gel P-10 low pressure column chromatography to obtain a uniform mixture of dodecanose. The mixture was analyzed by Strong Anion Exchange High Performance Liquid Chromatography SAX-HPLC) to obtain a uniform charge of pure dodecane. The oligosaccharides were completely degraded by heparanase Ⅰ, Ⅱ and Ⅲ, and the disaccharide composition was analyzed. The structural sequence was confirmed by electrospray ionization collision-induced tandem mass spectrometry (ES-CIDMSn). As a result, a pure and uniform charge of dodecane consisting of ΔUA-GlcNAc (Ⅳ-A) and ΔUA-GlcNS (Ⅳ-S) was isolated from the HS oligosaccharide mixture at a relative ratio of 5: 1, confirmed by ES-CID-MSn analysis, the structural sequence is ΔUA-GlcNAc-GlcA-GlcNS-GlcA-GlcNAcGlcA-GlcNAc-GlcA-GlcNAc-GlcA-GlcNAc. Conclusions In this experiment, a kind of dodecane was prepared from HS, and its structure was analyzed. This laid the foundation for the study of the structure-activity relationship of this oligosaccharide.