LINKAGE ANALYSIS OF HUMAN MILK FRACTION 2

母乳组分 2 的连锁分析

• 批准号: 8170801
• 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/8170801
• 关键词: Acetates; Acetic Acids; Amino Sugars; Blood capillaries; Computer Retrieval of Information on Scientific Projects Database; Funding; Gases; Grant; Human Milk; Hydrolysis; Hydroxyl Radical; Institution; Mass Spectrum Analysis; Oligosaccharides; Phase; Preparation; Ramp; Research; Research Personnel; Resources; Silicon Dioxide; Source; Sugar Alcohols; Temperature; United States National Institutes of Health; Water; acetic anhydride; capillary; detector; genetic linkage analysis; pyridine; sugar

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. Preparation of Partially Methylated Alditol Acetates (PMAAs) For the determination of glycosyl linkages, PMAAs were prepared from the permethylated oligosaccharides (remainder from oligosaccharide profiling). Briefly, permethylated oligosaccharides were hydrolyzed with HCl/water/acetic acid (0.5:1.5:8, by vol.) at 80oC for 18h, followed by reduction with NaBD4. The latter-freed hydroxyls after hydrolysis were acetylated with acetic anhydride/pyridine (1:1, v/v) at 100 ¿C for 15 min. Gas Chromatograph-Mass Spectrometry (GC-MS) The PMAAs for neutral sugars were analyzed on an Agilent 7890A GC interfaced to a 5975C MSD equipped with a Rtx-2330 column, whereas, the PMAAs for amino sugars was analyzed on a Hewlett Packard (HP) 5890 GC interfaced to a 5970 installed with a 30-m EC 1 bonded phase fused silica capillary column (Altech). The GC conditions were the following: Agilent 7890A, Initial oven temp was 80oC, which was held for 2 min, ramped to 170oC at 30oC/min, held for 0 min, and finally ramped to 235oC at 4oC/min and held for 20 min; HP 5890 GC, oven temperature, 140¿C (2.0¿C/min) ¿ 220¿C (20¿C/min) ¿ 300¿C (20¿C/min); detector temperature, 280 ¿C; inlet temperature, 250 ¿C.

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。 制备部分甲基化乙醇乙酸醛醇（PMAA） 为了测定糖基链接，从氯二基化的寡糖（寡糖分析）制备了PMAA。简而言之，将二甲基化的寡糖用Hcl/Water/乙酸（0.5：1.5：8，by Vol。）在80oC下水解为18H，然后用NABD4还原。水解后后来的羟基用乙酰用乙酰酸酐/吡啶（1：1，v/v）在100°C下乙酰化15分钟。 气相色谱 - 质量光谱法（GC-MS） 在配备了RTX-2330列的5975C MSD的敏捷的7890A GC上分析了中性糖的PMAA，而在Hewlett Packard（HP）5890 GC上与5970 capere casee capiene Forech capered Fiele copered Fiele copere fore caper copere copere fore caper caper copere fore caper caper capere copere copere copere copere copere copere fore and copere caper capere copere fore capere fore则分析了氨基糖的PMAAS。 GC条件如下：Agilent 7890a，初始烤箱温度为80oC，保持2分钟，以30oc/min的速度升至170oC，保持0分钟，最终以4oc/min的速度升至235oC，并保持20分钟； HP 5890 GC，烤箱温度，140€c（2.0€c/min）€220�C（20€c/min）€300¿C（20€c/min）;探测器温度，280®;入口温度，250�C。