Development of Recovering Equipment of Free Oligosaccharides Chemically Converted from Fluorescent Labeled Glycans by Hydrogen Peroxide and Its Application of Two-dimensional Liquid Chromatography

本研究探讨了化学再生的游离寡糖从他们的Fuglecent衍生物制备还原胺与各种芳香胺。将4-氨基苯甲酸乙酯（p-ABEE）、2-氨基苯甲腈（o-ABN）、4-氨基苯甲腈（p-ABN）、7-氨基-4-甲基香豆素（AMC）、2-氨基苯甲酸（o-ABA）、2-氨基苯甲酰胺（o-ABAD）、2-氨基吡啶（AP）和8-氨基萘-1，3，6-三磺酰胺（ANTS）的麦芽糖衍生物在30℃下与过氧化氢/乙酸水溶液一起孵育。从p-ABEE、p-ABN和AMC衍生物中回收麦芽糖相当好，得到约90%的麦芽糖。从邻位取代苯胺（o-ABA、o-ABAD和o-ABN）衍生物中回收麦芽糖的回收率为5- 40%，但从AP和ANTS衍生物中不能回收麦芽糖。然而，用溴化氰预先处理AP衍生物能够以高产率再生麦芽糖。作为一个应用程序，从一些糖蛋白分离的酰胺柱上的p-ABEE-标记的N-聚糖，通过转换峰成分，其AP衍生物通过自由基和随后的映射，通过反相色谱法进行鉴定。

This study examined chemical regeneration of free oligosaccharides from their Fuorescent derivatives prepared by reductive amination with various aromatic amines. Maltose derivatives of ethyl 4-aminobenzoate (p-ABEE), 2-aminobenzonitrile (o-ABN), 4-aminobemonitrile (p-ABN), 7-amino-4-methylcoumarin (AMC), 2-aminobenzoic acid (o-ABA), 2-aminobenzamide (o-ABAD), 2-aminopyridine (AP), and 8-aminonaphthalene-1,3,6-trisulfonare (ANTS) were incubated at 30℃ with an aqueous solution of hydrogen peroxide/acetic acid. Recoveries of maltose from p-ABEE, p-ABN, and AMC derivatives were fairly good and gave approximately 90% of maltose. Recoveries of maltose from its o-substituted aniline (o-ABA, o-ABAD, and o-ABN) derivatives were 5-40%, but maltose was unrecoverable from AP and ANTS derivatives. Nevertheless, prior treatment of an AP derivative with cyanogen bromide enabled the regeneration of maltose in high yields. As an application, p-ABEE-labeled N-glycans from some glycoproteins separated on an amide column were identified by converting peak components to their AP derivatives via free saccharides and following mapping by reversed phase chromatography.