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.

通过以下方法分析样品的单糖组成： 制备甲基糖苷的 TMS 衍生物，然后 GC-MS 分析。 将样品酸水解，然后进行 甲醇分解，然后用吡啶和乙酸进行 N-乙酰化 酸酐（用于检测氨基糖）。 当时的样品是 经过 Tri-Sil 处理。 TMS 甲基糖苷的 GC-MS 分析 在与 5970 MSD 连接的 Hewlett-Packard 5890 GC 上执行 30米DB1熔融石英毛细管柱。 样品是 使用 Hakomori 甲基化进行糖基连接分析 程序随后进行组合 GC-MS 分析。 当时的样品是 使用2M三氟乙酸进行酸水解，以实现 所有糖苷键的断裂。 糖基残基是 用 NaBD4 还原，然后用乙酸乙酰化 酸酐/吡啶。 所得部分甲基化糖醇 在 30 米 Supelco 2330 熔融石英毛细管上分析醋酸盐 使用 Hewlett-Packard 5890 GC 与 5970 联用，通过 GC-MS 分析色谱柱 默沙东。