GLYCOSYL COMPOSITION ANALYSIS OF TWO SAMPLES

两个样品的糖基组成分析

• 批准号: 8170783
• 负责人: Parastoo Azadi
• 金额: $ 0.13万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170783
• 关键词: Acetylation; Acids; Aliquot; Amino Sugars; Blood capillaries; Computer Retrieval of Information on Scientific Projects Database; Detection; Funding; Glycosides; Grant; Hour; Inositol; Institution; Mass Fragmentography; Methanol; Methods; Monosaccharides; Procedures; Research; Research Personnel; Resources; Sampling; Silicon Dioxide; Source; Tube; United States National Institutes of Health; acetic anhydride; capillary; pyridine

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Glycosyl composition analysis was performed by combined gas chromatography/mass spectrometry (GC/MS) of the per-O-trimethylsilyl (TMS) derivatives of the monosaccharide methyl glycosides produced from the sample by acidic methanolysis. A 200ul aliquot was taken from the sample and added to separate tubes with 40ug of Inositol as the internal standard. Methyl glycosides were then prepared from the dry sample following the mild acid treatment by methanolysis in 1 M HCl in methanol at 80¿C (16 hours), followed by re-N-acetylation with pyridine and acetic anhydride in methanol (for detection of amino sugars). The sample was then per-O-trimethylsilylated by treatment with Tri-Sil (Pierce) at 80¿C (0.5 hours). These procedures were carried out as previously described in Merkle and Poppe (1994) Methods Enzymol. 230: 1-15; York, et al. (1985) Methods Enzymol. 118:3-40. GC/MS analysis of the TMS methyl glycosides was performed on an AT 6890N GC interfaced to a 5975B MSD, using a Supelco EC-1 fused silica capillary column (30m ¿ 0.25 mm ID).

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以出现在其他 CRISP 条目中 列出的机构是。 对于中心来说，它不一定是研究者的机构。 通过气相色谱/质谱联用 (GC/MS) 对通过酸性甲醇分解样品产生的单糖甲基糖苷的全 O-三甲基甲硅烷基 (TMS) 衍生物进行糖基组成分析。 从样品中取出 200ul 等分试样，加入装有 40ug 肌醇作为内标的单独试管中，然后通过在 80° 的 1 M HCl 甲醇溶液中进行弱酸处理，从干燥样品中制备甲基糖苷。 C（16 小时），然后用甲醇中的吡啶和乙酸酐进行重新 N-乙酰化（用于检测氨基糖），然后通过在 80° 下用 Tri-Sil (Pierce) 处理来对样品进行全 O-三甲基甲硅烷基化。 C（0.5小时）。这些程序如先前在Merkle和Poppe（1994）Methods Enzymol.230：1-15；York等人（1985）Methods Enzymol.118：3-40中所述进行。 TMS 甲基糖苷的分析是在连接 5975B MSD 的 AT 6890N GC 上进行的，使用Supelco EC-1 熔融石英毛细管柱（30m ¿ 0.25 mm ID）。