Core B: Metabolomics Core

该核心将提供混合(靶向、非靶向)代谢组学平台，以支持 根据POI研究的需要，对红细胞和其他生物样本进行化学分析。 中心平台由双色谱-高分辨质谱仪组成 再加上一系列数据提取算法，提供了对 高达90,000个高质量精度(5 Ppm)m/z要素，超过10,000个 人体样本中存在的化学物质。有针对性的分析允许直接测量 超过300种具有已知身份和能力的化学品，可检测不明身份和 对结果有影响的未知化学物质。对数以千计的 样本显示，该方法测量的离子与超过一半的 人体代谢图中的代谢物.用共洗脱法选定的身份确认 而MS/MS表明，该方法提供了有效的全球新陈代谢调查， 捕获KEGG人类代谢途径154条途径中的146条代谢产物。 该方法可能对当前的提案有用，还检测到数千个 来自饮食、宿主微生物群、感染剂和环境的化学物质 暴露，这可能导致受者对输血的易感性发生变化 存储的RBC。严格遵守标准操作程序，使用专门的 具有内部标准和外部汇集参考样品的仪器将提供 支持代谢组范围的关联研究(MWAS)以识别两者的能力 红细胞中与特定临床相关的内源性和外来化学物质 和/或拟议研究中的功能结果。样品分析一式三份 从而确定每个样本中每个m/z特征的变异系数。 数据提取参数的选择允许更宽松的数据提取标准 基于发现的分析和使用稳定同位素的更严格的目标标准 稀释和多离子监测，以检验特定的假设。对于未知的和 未知化学物质，由准确质量m/z组成的质谱学数据，保留 时间和离子强度的相对量化与基于数据的搜索一起使用 推定的化学鉴定。数据依赖离子解离(MS/MS和MS“) 对有代表性的或选定的样品进行分析，以确认 靶向代谢物和/或高丰度m/z特征。

This core will provide a hybrid (targeted, non-targeted) metabolomics platform to support chemical profiling of RBC and other biological samples as needed by the POI studies. The central platform consists of dual chromatography-high-resolution mass spectrometry coupled with a series of data extraction algorithms that provide relative quantification of up to 90,000 high-mass accuracy (5 ppm) m/z features, representing in excess of 10,000 chemicals present in human samples. Targeted analysis allows direct measurement of over 300 chemicals with known identity and capabilities to detect unidentified and unknown chemicals contributing to outcomes. Experience with analyses of thousands of samples shows that the method measures ions that match more than half of the metabolites in human metabolic charts; selected confirmation of identities by co-elution and MS/MS shows that the approach provides an effective global survey of metabolism, capturing metabolites in 146 out of 154 pathways in KEGG human metabolic pathways. Potentially useful for the current proposal, the approach also detects thousands of chemicals derived from diet, the host microbiome, infectious agents and environmental exposures, which could contribute to variation in recipient susceptibility to transfusions of stored RBCs. Rigorous adherence to standard operating procedures using dedicated instruments with internal standards and external pooled reference samples will provide the capability to support metabolome-wide association studies (MWAS) to identify both endogenous and foreign chemicals in RBCs that are associated with specific clinical and/or functional outcomes in the proposed studies. Samples are analyzed in triplicate so that coefficients of variation are determined for each m/z feature in each sample. Selection of data extraction parameters allows more liberal data extraction criteria for discovery-based analysis and more rigorous targeted criteria using stable isotope dilution and multiple ion monitoring to test specific hypotheses. For unknown and unidentified chemicals, mass spectral data consisting of accurate mass m/z, retention time and relative quantification as ion intensity are used with data-based searching for putative chemical identification. Data dependent ion-dissociation (MS/MS and MS") analyses are provided on representative or selected samples to confirm identities of targeted metabolites and/or high abundance m/z features.