Metabolomics Core

.PROJECT摘要(请参阅说明)： 代谢组学核心提供了一种工具来确定糖尿病及其并发症的转基因小鼠模型的代谢物和激素的表型。代谢组学核心提供的服务包括从组织和血浆中提取、纯化、衍生化和仪器分析代谢物、激素和选定的酶活性。这些方法已经被扩展到考虑到研究人员可以提供的最少量的组织(-10到25毫克，以及-5到25 pi的血浆体积)。 该设施配备了一系列最先进的仪器，包括GC-MS、LC/MS/MS、高效液相色谱和核磁共振，为代谢物浓度分析和同位浓缩提供了灵活性和资源。使用COBAS MIRA系统获得血清、血浆和尿液的代谢面板，包括Chem 7、肝功能测试、脂谱和二价阳离子。我们已经优化了COBAS MIRA分析系统，用于分析小鼠样本，所需的血清体积平均为-20pi，用于4种测试的任意组合。这些等离子体化学分析填补了对小鼠研究的非常高的需求，这反映在我们上个资助期的工作流程中。GC/MS和LC/MS/MS分析提供了脂肪酸代谢物的完整链长分布，包括游离脂肪酸、脂肪酰-CoA酯、二酰甘油和神经酰胺。其他提供的和需求量很大的LC/MS/MS包括用于代谢通量测量的葡萄糖代谢中间体，以及用于肾脏清除的肌酐。 代谢组学核心是一个资源实验室，用于分析耶鲁大学MMPC综合生理学核心实验过程中产生的样本，并向其他外部研究人员提供相同的分析阵列，以帮助进行糖尿病转基因小鼠模型的表型分析，并根据国家MMPC执行委员会制定的政策对糖尿病样本进行收费服务分析，旨在为NIH资助的用户提供与商业供应商相比大幅节省的费用。核心有一个积极的研究和开发计划，以开发新的分析方案，以解决糖尿病、肥胖症和相关并发症的生物化学方面的新和令人兴奋的发展

.PROJECT SUMMARY (See instructions): The Metabolomics Core provides a facility to determine the phenotype of metabolite and hormone profiles of transgenic mouse models of diabetes and its complications. The services provided by the Metabolomics Core include the extraction, purification, derivatization, and instrumental analysis of metabolites, hormones, and selected enzyme activities from tissues and plasma. The methods have been scaled to account for the minimal amounts of tissues (-10 to 25 mg, and plasma volumes of - 5 to 25 pi) that can be provided by investigators.. The facility is equipped with an array of state-of-the-art instrumentation, including GC-MS, LC/MS/MS, HPLC, and NMR, providing the flexibility and resources for analyses of metabolite concentrations as well as sotopic enrichments. Metabolic panels of serum, plasma, and urine, including Chem 7, liver function tests lipid profiles, and divalent cations, are obtained using the COBAS MIRA system. We have optimized the COBAS Mira analytical system for analysis of mouse samples such that required volumes average -20 pi of serum for any combination of 4 tests. These plasma chemistry assays fill a need with very high demand for mouse studies, as reflected in our work flow over the last funding period. GC/MS and LC/MS/MS analyses provide complete profiles of chain length distribution of fatty acid metabolites, included free fatty acids, fatty acyl-CoA esters, diacylglycerols, ceramides. Other LC/MS/MS that are offered and are in high-demand include intermediates of glucose metabolism for metabolic flux measurements, and creatinines for renal clearance. The Metabolomics Core is a resource lab for the analysis of samples generated during the course of experiments of the Integrative Physiology Core at the Yale MMPC, and also provides the same array of assays to other outside investigators to assist in phenotyping of transgenic mouse models of diabetes Samples are analyzed on a fee-for-service basis with the charge per assay based on policy determined by the National MMPC Executive Committee, designed to provide substantial savings to NIH-funded users compared to commercial vendors. The Core has an active Research and Development Program to develop new analytical protocols to address new and exciting developments in the biochemistry of diabetes, obesity, and associated complications