TECHNOLOGY NMR SPECTROSCOPY

作为梅奥临床综合代谢组学资源核心(MCCMRC)的组成部分，核磁共振 代谢组学核心使用核磁共振对代谢组谱进行有针对性的分析 (核磁共振)谱学。除了建立标准核磁共振高通量1H和31P核磁共振 代谢组学分析，核心将实施其独特的18O标记和18O辅助31P的方法 研究磷代谢动力学的核磁共振波谱和碳同位素异构体的~(13)C核磁共振 代谢网络中的交换，这是一个集成在质谱学核心中的过程。这一集成 组织提取物的定性和定量分析(1H和31P核磁共振)、稳定同位素示踪的方法 (13C和18O辅助的31P核磁共振波谱)和活体波谱(1H，31P体积选择核磁共振 光谱学、核磁共振光谱成像)将包括对人类和动物样本的分析(例如， 全血、血浆、脑脊液、尿、心、肝、脑、肾)和体内完整的器官(脑、 心脏、肾脏、肌肉)。此外，我们将实施应激代谢测试方案，在 在运动之后。核磁共振代谢组学核心由分析核磁共振设备支持，该设备具有5个最先进的 300兆赫、500兆赫、600兆赫、700兆赫和大口径700兆赫核磁共振波谱仪，配备 经验丰富的核磁共振光谱学家。我们建议获得一个高灵敏度的600 MHz BB低温探头， 高通量样品处理机和额外的技术人员，以扩大核心能力和分析 1H的能力至少是三倍，31P和13C的核磁共振能力至少是4-9倍(每天-25到100个样品)， 为梅奥诊所内外更多的调查人员提供服务。核心将会 实施标准化的代谢方案和核磁共振数据采集、处理、 代谢物鉴定、生物信息途径和通量分析与解释。核心将会发展 确保具有成本效益的系列样品分析和质量控制的适当工作流程。这个 核心员工将与MCCMRC管理层合作，实施一致的格式来存放RAW、 根据数据共享计划对数据进行处理和分析。目前的核磁共振代谢核心 协助20多名NIH在心血管、衰老、糖尿病、癌症和神经退行性疾病方面的研究人员 疾病，以及再生和个体化医学领域。我们计划制作这个节目 在五年内自给自足。因此，该应用程序的总体目标是巩固和扩展核磁共振 提供代谢核心设施和服务，并向其他国家机构提供我们的专业知识 作为区域枢纽，推进基于稳定同位素和核磁共振的翻译和临床代谢组学。

As an integral part of Mayo Clinic Comprehensive Metabolomics Resource Core (MCCMRC) the NMR Metabolomics core provides targeted analysis of metabolomic profiles using Nuclear Magnetic Resonance (NMR) Spectroscopy. In addition to establishing standard NMR high-throughput 1H and 31P NMR metabolomics analyses, the core will implement, its unique methods of 18O labeling and 18O-assisted 31P NMR spectroscopy for studies of phosphometabolite dynamics and 13C NMR for carbon isotopomer exchange in metabolic networks, a procedure that is integrated in Mass Spectrometry core. This integrated approach of qualitative and quantitative analysis of tissue extracts (1H and 31P NMR), stable isotope tracing (13C and 18O-assisted 31P NMR spectroscopy) and in vivo spectroscopy (1H, 31P volume selective NMR spectroscopy, NMR spectroscopic imaging) will include the analysis of human and animal samples (e.g., whole blood, plasma, cerebrospinal fluid, urine, heart, liver, brain, kidney) and in vivo intact organs (brain, heart, kidney, muscle). In addition, we will implement protocols for stress metabolomic testing before and after exercise. The NMR Metabolomics core is supported by Analytical NMR Facility that has 5 state-of-theart 300 MHz, 500 MHz, 600 MHz, 700 MHz and wide bore 700 MHz NMR spectrometers and is staffed with experienced NMR spectroscopists. We propose the acquisition of a high sensitivity 600 MHz BB cryoprobe, high-throughput sample handler and additional technical staff to expand the core capacity and analytical capabilities at least threefold for 1H and 4-9 fold for 31P and 13C NMR (from -25 to 100 samples/day) and to offer services and to a larger number of investigators inside and outside Mayo Clinic. The core will implement standardized metabolomic protocols and methods of NMR data acquisition, processing, metabolite identification, bioinformatic pathway and flux analysis and interpretation. The core will develop appropriate workflow that will ensure cost-effective analysis of series of samples, and quality control. The core staff will work with the MCCMRC Management to implement consistent formats to deposit raw, processed and analyzed data according to the data sharing plan. The NMR Metabolomic core currently assists over 20 NIH supported investigators in cardiovascular, aging, diabetes, cancer and neurodegenerative disease, and regenerative and individualized medicine areas. We plan to make the program self-sustaining in five years. Thus the overall goal of this application is to consolidate and expand the NMR Metabolomic core facility and services it provides and to offer our expertise to other national institutions serving as a regional hub to advance stable isotope and NMR based translational and clinical metabolomics.