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).

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 糖基组成分析是通过酸性甲基溶解的样品产生的单糖甲基甲基糖苷的每甲基甲硅烷基（TMS）衍生物的共同气相色谱/质谱法（GC/MS）进行的。 从样品中取一个200ul等分试样，并将40倍肌醇作为内标的分开试管。然后，通过在80 ch（16小时）的甲醇中甲烷溶液在1 M HCL中进行甲烷溶液处理后，从干样样品中制备甲基糖苷，然后用吡啶定和乙醇中的乙醇中的乙酰化重新 - N-乙酰化（用于检测氨基糖）。然后，通过在80°C（0.5小时）处理三米（Pierce）处理样品，每三甲基硅烷基硅烷基硅烷基硅烷基硅烷基。这些程序是如前所述的Merkle和Poppe（1994）方法进行的。 230：1-15;约克等。 （1985）方法酶。 118：3-40。使用Supelco EC-1 Fused Silica Capillary柱（30m€0.25 mm ID），在6890N的GC上对TMS甲基糖苷进行GC/MS分析。