GLYCOSYL COMPOSITION ANALYSIS OF SACCHARIDE

糖的糖基组成分析

• 批准号: 6653596
• 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/6653596
• 关键词: biological products; biomedical resource; carbohydrates; oral pharyngeal; spectrometry

Glycosyl-linkage analysis was performed by preparing permethylated alditiol acetates using the Hakomori procedure. The samples were dissolved in DMSO, DMSO anion was added, and the samples were stirred at room temperature for 2 h. An excess of methyliodide was added, and the samples were stirred again from 2 to 4 h at room temperature. Excess methyliodide was removed by evaporation, and the samples were purified using a C18 SepPak cartridge. The resulting samples were hydrolyzed in 2 M TFA, and the monosaccharides were reduced to their alditiols and N-acetylated using acidic anhydride in pyridine. The resulting acetates were then analyzed by GC-MS using a 30 mm SP2330 column from Supelco for the neutral sugars and using a DB1 column from J&W Scientific for the methylated amino sugars.

糖基连接分析通过制备全甲基化的 使用Hakomori方法制备乙酸醛地替醇。 将样品 溶解在DMSO中，加入DMSO阴离子，并搅拌样品 在室温下反应2小时。加入过量的碘甲烷， 在室温下再次搅拌样品2 - 4小时。 通过蒸发除去过量的甲基碘，并将样品在室温下蒸发。 使用C18 SepPak柱纯化。 将所得样品 在2 M TFA中水解，并将单糖还原至其 醛硫醇并在吡啶中使用酸酐进行N-乙酰化。 的 然后使用30 mm SP2330通过GC-MS分析所得乙酸酯 中性糖使用Supelco的DB 1柱， J&W Scientific的甲基化氨基糖。