GLYCOSYL COMPOSITION ANALYSIS OF TWO SAMPLES BY GC-MS

通过 GC-MS 分析两个样品的糖基组成

• 批准号: 8363062
• 负责人: Parastoo Azadi
• 金额: $ 0.17万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363062
• 关键词: Acetylation; Amino Sugars; Blood capillaries; Detection; Funding; Gases; Glycosides; Grant; Guanine + Cytosine Composition; Hexanes; Inositol; Methanol; Methods; National Center for Research Resources; Principal Investigator; Procedures; Reagent; Research; Research Infrastructure; Resources; Sampling; Series; Silicon Dioxide; Source; Technology; Tube; United States National Institutes of Health; acetic anhydride; capillary; cost; pyridine

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Glycosyl composition by GC-MS The sample was transferred to a pre-rinsed screw-cap tube, added with 3 ¿g of inositol as an internal standard, and lyophilized. Methyl glycosides then were prepared from the dried sample by methanolysis with 1 M HCl in methanol at 80¿C overnight followed by re-N-acetylation with pyridine and acetic anhydride in methanol (for detection of amino sugars). The methyl glycosides of the sample were per-O-trimethylsilylated (TMS) with a Tri-Sil reagent (Thermo Scientific) at 80¿C for 0.5 h. These procedures were carried out as described previously in Merkle and Poppe (1994) Methods Enzymol. 230: 1-15; York, et al. (1985) Methods Enzymol. 118:3-40. Analysis of the TMS methyl glycosides was performed on a Hewlett Packard Series II 5890 gas chromatograph equipped with a Supelco EC-1 fused silica capillary column (30m ¿ 0.25 mm ID) and interfaced to a Hewlett Packard 5970 MSD. Since the absolute content of the analytes was expected to be low, the final derivatized sample was dissolved in a small volume of hexane and had to be injected manually.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供 该子项目的主要支持。 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源的子项目可能列出的总成本。 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 GC-MS 测定的糖基组成 将样品转移至预冲洗的旋盖管中，添加 3 ¿ g 肌醇作为内标，然后通过在 80° 下用 1 M HCl 的甲醇溶液进行甲醇分解，从干燥的样品中制备冻干的甲基糖苷。 C过夜，然后用甲醇中的吡啶和乙酸酐进行再N-乙酰化（用于检测氨基糖）。在30℃下用Tri-Sil试剂（Thermo Scientific）对样品的甲基糖苷进行全O-三甲基甲硅烷基化（TMS）。 80° C 0.5 小时，如先前 Merkle 和 Poppe (1994)Methods Enzymol.230:1-15；York 等人 (1985)Methods Enzymol.118:3-40 中所述进行。甲基糖苷分析是在配有 Supelco EC-1 融合的 Hewlett Packard Series II 5890 气相色谱仪上进行的硅胶毛细管柱 (30m ¿ 0.25 mm ID) 并连接到 Hewlett Packard 5970 MSD 由于预计分析物的绝对含量较低，因此最终的衍生样品溶解在少量己烷中，并且必须手动注入。 。