Core B: Metabolomics Core

该核心将提供一个混合（靶向，非靶向）代谢组学平台，以支持 POI研究所需的RBC和其他生物样本的化学分析。 中心平台由双重色谱-高分辨质谱组成 再加上一系列的数据提取算法， 高达90，000个高质量精度（5 ppm）m/z特征，代表超过10，000个 人体样本中的化学物质有针对性的分析可以直接测量 超过300种化学品具有已知的特性和检测能力， 未知的化学物质对结果的影响。分析数千种 样品表明，该方法测量的离子匹配超过一半的 人体代谢图中的代谢物.共洗脱法鉴定的选择性确证 MS/MS显示该方法提供了有效的代谢全球调查， 在KEGG人体代谢途径的154条途径中的146条中捕获代谢物。 对于当前的提案可能有用，该方法还检测了数千个 来自饮食、宿主微生物组、传染源和环境的化学品 暴露，这可能导致受体对输血敏感性的变化， 储存RBC。严格遵守标准操作程序， 具有内标物和外部混合参考样品的仪器将提供 支持全代谢组关联研究（MWAS）的能力， 红细胞中的内源性和外源性化学物质与特定临床 和/或功能结局。一式三份分析样品 从而确定每个样本中每个m/z特征的变化系数。 数据提取参数的选择允许更自由的数据提取标准， 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，以测试特定的假设。对于未知和 未鉴别化学品，质谱数据包括准确的质量m/z，保留时间 时间和离子强度的相对定量与基于数据的搜索一起使用， 推定化学鉴定数据依赖性离子解离（MS/MS和MS”） 对代表性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.