Hou Zhiqiang, Wang Wenkang, Wang Yuying, Chen Shiguo, Ye Xingqian
College of Biosystems Engineering and Food Science, National-Local Joint Engineering Laboratory of Intelligent Food Technology and Equipment, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang Engineering Laboratory of Food Technology and Equipment, Zhejiang University, Hangzhou 310058, China.
College of Biosystems Engineering and Food Science, National-Local Joint Engineering Laboratory of Intelligent Food Technology and Equipment, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang Engineering Laboratory of Food Technology and Equipment, Zhejiang University, Hangzhou 310058, China; College of Agriculture & Biotechnology, Zhejiang University, Hangzhou 310058, China.
Int J Biol Macromol. 2025 Feb;291:138535. doi: 10.1016/j.ijbiomac.2024.138535. Epub 2024 Dec 9.
Glycosaminoglycans of marine origin have exceptional biological activities, but rapid elucidation strategies for precise structures are still lacking. In this study, the optimal conditions for deacetylation of glycosaminoglycans were optimized first. The hydrazinolysis time of 24 h was determined as the final condition, oligosaccharides with a degree of polymerization of 2-14 are mainly produced after hydrazinolysis. Then mixed oligosaccharides generated by deacetylation-deamination cleavage of polysaccharides were detected with hydrophilic interaction chromatography with tandem mass spectrometry (HILIC-MS/MS), and the total ion chromatography showed excellent resolution. The mass spectrometry data were automatically processed by GlycReSoft, and the corresponding matching results were obtained according to the set parameters. A small amount of keratan sulfate exists in oligosaccharides of Teuthida and Salmo salar, and ion peaks corresponding to IdoA-GalNAc4S exist in oligosaccharides of Scophthalmus maximus and Salmo salar. The results of the manual analysis of the mass spectrometry data verified that the HILIC-MS/MS combined with automatic analysis by software adopted in this study could achieve accurate matching and matching of mixed oligosaccharide components. The establishment of a rapid and accurate analysis strategy for the structure of glycosaminoglycans in this study will help the development of its application and standardized industrial production.
海洋来源的糖胺聚糖具有特殊的生物活性，但仍缺乏用于精确结构快速解析的策略。在本研究中，首先对糖胺聚糖脱乙酰化的最佳条件进行了优化。确定24小时的肼解时间为最终条件，肼解后主要产生聚合度为2 - 14的寡糖。然后采用亲水相互作用色谱-串联质谱（HILIC-MS/MS）对多糖脱乙酰化-脱氨裂解产生的混合寡糖进行检测，总离子色谱显示出优异的分辨率。质谱数据由GlycReSoft自动处理，并根据设定参数获得相应的匹配结果。柔鱼和大西洋鲑的寡糖中存在少量硫酸角质素，大菱鲆和大西洋鲑的寡糖中存在与艾杜糖醛酸-4-硫酸半乳糖胺对应的离子峰。对质谱数据的手动分析结果证实，本研究采用的HILIC-MS/MS结合软件自动分析能够实现混合寡糖成分的准确匹配和比对。本研究中糖胺聚糖结构快速准确分析策略的建立将有助于其应用开发和标准化工业生产。
