PROTON NMR OF 1 SAMPLE

1 个样品的质子核磁共振

• 批准号: 8361833
• 负责人: Parastoo Azadi
• 金额: $ 0.18万
• 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: 2011-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361833
• 关键词: Acetates; Acetone; Aliquot; Blood capillaries; COSY; Chemicals; Deuterium; Dimethyl Sulfoxide; Electrons; Freeze Drying; Funding; Grant; Infusion procedures; Left; Mass Fragmentography; Measures; Methods; Methylation; NMR Spectroscopy; NOESY; National Center for Research Resources; Phase; Polymers; Principal Investigator; Protons; Relative (related person); Research; Research Infrastructure; Resources; Running; Sampling; Silicon Dioxide; Sodium Hydroxide; Solutions; Source; Spectrometry, Mass, Electrospray Ionization; Sugar Alcohols; TOCSY; Trifluoroacetic Acid; Tube; United States National Institutes of Health; Water; acetic anhydride; base; capillary; cost; detector; genetic linkage analysis; ionization; methyl iodide; pyridine; seal

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. Methods: NMR Spectroscopy The sample was deuterium exchanged by dissolving in D2O and lyophilization. The sample was dissolved in 80 ¿L D2O and placed in a 3 mm Shigemi NMR tube. 1-D Proton, TOCSY and NOESY NMR spectra, run with water presaturation, and gradient enhanced COSY and HSQC spectra were acquired on a Varian Inova-800 MHz spectrometer at 298 K (25 ¿C). Chemical shifts were measured relative to internal acetone (¿H=2.218 ppm, ¿C=33.00 ppm). Permethylation For glycosyl linkage analysis, the sample was permethylated, depolymerized, reduced, and acetylated; and the resultant partially methylated alditol acetates (PMAAs) analyzed by gas chromatography-mass spectrometry (GC-MS) as described by York et al (1985) Methods Enzymol. 118:3-40. Initially, an aliquot of the sample was freeze-dried and suspended in about 200 ¿l of dimethyl sulfoxide. The sample was then permethylated by the method of Ciukanu and Kerek (1984) Carbohydr. Res. 131:209-217 (treatment with sodium hydroxide and methyl iodide in dry DMSO). The sample was subjected to the NaOH base for 10 minutes then methyl iodide was added and left for 40 minutes. The base was then added and left for 10 minutes and more methyl iodided was added and left for 40 minutes. The addition of more methyl iodide and NaOH base were to insure complete methylation of the polymer. Linkage analysis Following sample workup, the permethylated material was hydrolyzed using 2 M trifluoroacetic acid (2 h in sealed tube at 121¿C), reduced with NaBD4, and acetylated using acetic anhydride/pyridine. The resulting PMAAs were analyzed on an Agilent 7890A GC interfaced to a 5975C MSD (mass selective detector, electron impact ionization mode); separation was performed on a 30 m EC-1 bonded phase fused silica capillary column. ESI Mass spectrometry The permethylated sample was analyzed by direct infusion in 50 % MeOH solution on a ThermoFisher LTQ. Experimental details are available upon request.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 方法： NMR光谱 样品是通过溶解在D2O和冻干中交换的氘。将样品溶解在80`l d2O中，并放入3 mM Shigemi NMR管中。 1-D质子，Tocsy和Noesy NMR光谱，具有水的天度性运行，并在298 K（25»C）的Varian Inova-800 MHz光谱仪上获得了梯度增强的舒适和HSQC光谱。相对于内部丙酮测量化学位移（H = 2.218 ppm，�C= 33.00 ppm）。 苄苄氨基化 为了进行糖基链接分析，样品被氯化，沉积，还原和乙酰化；如约克等人（1985年）方法所述，由气相色谱 - 质谱法（GC-MS）分析的部分甲基化乙醇（PMAA）（PMAA）。 118：3-40。 最初，将样品的等分试样冷冻干燥，并悬浮在约200 l二甲基亚硫氧化二甲基中。然后，通过Ciukanu和Kerek（1984）碳水化合物的方法将样品苄苄烯基化。 res。 131：209-217（在干燥DMSO中用氢氧化钠和碘化甲基处理）。将样品持续10分钟，然后添加碘化甲基并保留40分钟。然后添加底座并剩下10分钟，然后添加更多的碘化甲基碘化甲基，然后剩下40分钟。添加更多的碘化甲基和NaOH碱是为了确保聚合物的完全甲基化。 链接分析 样品工作后，使用2 M三氟乙酸（在121€c处密封的管中2小时）将苄苄化材料水解，用NaBD4还原，并使用乙酰酸酐/吡啶乙基化。在连接到5975C MSD的安捷伦7890A GC上分析了所得的PMAA（质量选择性检测器，电子影响电离模式）；分离在30 m的EC-1键相融合二氧化硅毛细管柱上进行。 ESI质谱法 通过在Thermofisher LTQ上的50％MEOH溶液中直接输注二苄甲基化的样品。可应要求提供实验细节。