N LINKED OLIGOSACCHARIDE PROFILING

N 连接寡糖分析

• 批准号: 6653592
• 负责人: ROBERTA K MERKLE
• 金额: $ 28.8万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6653592
• 关键词: bacteria; biological products; biomedical resource; carbohydrates; spectrometry

The samples were analyzed for monosaccharide composition by preparing the TMS derivatives of the methylglycosides followed by GC-MS analysis. The samples were acid hydrolyzed then subjected to methanolysis followed by N-acetylation with pyridine and acetic anhydride (for detection of amino sugars). The samples were then treated with Tri-Sil. GC-MS analysis of the TMS methylglycosides was performed on a Hewlett-Packard 5890 GC coupled to a 5970 MSD using a 30-meter DB1 fused silica capillary column. The samples were subjected to glycosyl-linkage analysis using the Hakomori methylation procedure followed by combined GC-MS analysis. The samples were then subjected to acid hydrolysis using 2 M trifluoroacetic acid to effect cleavage of all glycosidic linkages. The glycosyl residues were reduced with NaBD4 and then acetylated using acetic anhydride/pyridine. The resulting partially methylated alditol acetates were analyzed on a 30-meter Supelco 2330 fused silica capil lary column by GC-MS using a Hewlett-Packard 5890 GC coupled to a 5970 MSD.

通过通过 制备甲基糖苷的TMS衍生物，然后 GC-MS分析。 样品被酸水解，然后进行 甲烷溶解，然后用吡啶和乙酸N-乙酰化 酸酐（用于检测氨基糖）。 然后样品是 用Tri-Sil处理。 TMS甲基糖苷的GC-MS分析为 在惠普（Hewlett-Packard）上进行5890 GC，耦合到5970 MSD 30米DB1融合二氧化硅毛细管柱。 样品是 使用hakomori甲基化进行糖基 - 连接分析 程序随后进行了GC-MS分析。 然后样品是 使用2 M三氟乙酸进行酸水解作用 所有糖苷链接的切割。 糖基残基是 用NaBD4还原，然后使用乙酰化乙酰化 酸酐/吡啶。 产生的部分甲基化醛醇 在30米的supelco 2330融合二氧化硅Capil上分析了醋酸盐 GC-MS使用惠普5890 GC的喉柱，耦合到5970 MSD。