TECHNOLOGY NMR SPECTROSCOPY

作为马约临床综合代谢组学资源核心（MCCMRC）的组成部分， Metabolomics core使用核磁共振技术对代谢组学特征进行靶向分析 (NMR)谱除了建立标准的NMR高通量1H和31 P NMR 代谢组学分析，核心将实施，其独特的方法，18 O标记和18 O辅助31 P 磷代谢物动力学研究的NMR谱和碳同位素的~（13）C NMR 代谢网络中的交换，这是一个集成在质谱核心中的程序。这一综合 组织提取物的定性和定量分析方法（1H和31 P NMR），稳定同位素示踪 (13C和18 O辅助的31 P NMR光谱法）和体内光谱法（1H，31 P体积选择性NMR 光谱学，NMR光谱成像）将包括对人类和动物样品的分析（例如， 全血、血浆、脑脊髓液、尿、心脏、肝、脑、肾）和体内完整器官（脑， 心脏、肾脏、肌肉）。此外，我们将实施压力代谢组学测试的协议， 锻炼后。NMR代谢组学核心由分析NMR设施提供支持，该设施具有5个最先进的 300 MHz、500 MHz、600 MHz、700 MHz和宽口径700 MHz NMR光谱仪，并配备 经验丰富的核磁共振波谱仪。我们建议获得高灵敏度600 MHz BB冷冻探针， 高通量样品处理器和额外的技术人员，以扩大核心能力和分析 1H NMR的能力至少为3倍，31 P和13 C NMR的能力至少为4-9倍（每天约25至100个样品）， 为马约诊所内外的大量研究者提供服务。芯会 实施NMR数据采集、处理 代谢物鉴定、生物信息学途径和通量分析和解释。核心将发展 适当的工作流程，确保对一系列样品进行具有成本效益的分析和质量控制。的 核心员工将与MCCMRC管理层合作，实施一致的格式，以存款原始数据， 根据数据共享计划处理和分析数据。NMR代谢组学核心目前 协助20多名NIH支持的心血管、衰老、糖尿病、癌症和神经退行性疾病研究人员 疾病，再生和个性化医疗领域。我们计划让这个节目 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NMR 它提供代谢组学核心设施和服务，并向其他国家机构提供我们的专业知识 作为区域中心，推进基于稳定同位素和NMR的转化和临床代谢组学。

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.