TECHNOLOGY NMR SPECTROSCOPY

Mayo Clinic Rochester, Rochester MN

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Abstract

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.