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).

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 糖基组成分析进行了组合的气相色谱/质谱（GC/MS）的全-O-三甲基甲硅烷基（TMS）衍生物的单糖甲基糖苷的样品通过酸性甲醇分解。 从样品中取出200 μ 1等分试样，并加入到具有40 μ g肌醇作为内标的单独管中。然后，在温和的酸处理后，通过在80 ℃下在甲醇中的1 M HCl中甲醇分解（16小时），然后在甲醇中用吡啶和乙酸酐进行再N-乙酰化（用于检测氨基糖），从干燥样品制备甲基糖苷。 然后通过在80 ℃下用Tri-Sil（Pierce）处理（0.5小时）将样品全-O-三甲基甲硅烷基化。这些程序如先前在Merkle和Poppe（1994）Methods Enzymol. 230：1-15;约克等人（1985）酶学方法（Methods Enzymol. 118：3-40。TMS甲基糖苷的GC/MS分析在与5975 B MSD连接的AT 6890 N GC上进行，使用Supelco EC-1熔融石英毛细管柱（30 m ² 0.25 mm ID）。