Colorado PROFILES, The Colorado Clinical and Translational Sciences Institute (CCTSI)
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Integrated state-of-the-art LC-MS/NMR system for the University of Wisconsin


Collapse Biography 

Collapse Overview 
Collapse abstract
Funds are requested for the purchase of an integrated liquid-chromatography, mass spectrometer, and nuclear magnetic resonance (LC-MS/NMR) system. This system will be housed in the National Magnetic Resonance Facility at Madison (NMRFAM) located in the Biochemistry Department at the University of Wisconsin-Madison. The system will be installed in a room with tightly controlled temperature and humidity and with all needed utilities already in place. The system will be used to support ongoing, NIH-supported, research projects in four broad areas: (1) Analysis of metabolite mixtures (metabolomics). (2) Screening of drug-receptor (ligand- protein) interactions. (3) Screening of protein and nucleic acid samples by NMR for proper folding under various conditions and by MS for quality control (correct identity, labeling, and purity). (4) Elucidation of structures of natural products available only in small quantities. The LC-MS/NMR will have unique capabilities unmatched on this campus or at currently any other academic facility in the US. The system will have automated, multiple sample operation with temperature control of samples. It will have the ability to acquire ESI/APCI Q-TOF MS or MS/MS data on aliquots of samples (unfractionated or LC fraction) that can be investigated also by NMR spectroscopy. The NMR spectrometer will consist of a 600 MHz shielded magnet, a multinuclear console, and a cryogenic platform. Users will have the choice of two triple-resonance (1H or 13C observe, 13C or 15N decoupling, 2H lock) cryogenic probes: for adequate samples, a 5 mm probe will provide the highest sensitivity (300 ?L samples in tubes); for sample-limited situations, a 1.7 mm probe will provide superior performance (30 ?L samples in tubes). This NMR system will enable the economical screening of proteins and nucleic acids and investigations of natural products obtainable only in small quantities. The system will have a solid-phase extraction accessory capable of concentrating samples by up to 10-fold. A liquids handling system will transfer samples to the MS system or NMR tubes. A thermostatted sample changer will handle the automated screening of NMR samples in tubes. The system will have a software system for automated identification and quantification of metabolites and related biosynthetic pathways. NMRFAM staff members will maintain the instrumentation, assist in performing studies, and train users who desire hands-on experience how to use the system safely and properly. User fees will be levied to recover operating expenses. Scheduling will be by an existing web-based system, and oversight will be provided by existing local and external advisory committees. PUBLIC HEALTH RELEVANCE: The research supported by this instrumentation is expected to benefit public health because it aims to investigate metabolic markers of aging and various disease states and to determine the structures of natural products that are candidate new drugs and antibiotics. The instrumentation will enable more complete investigations of the structures of proteins and nucleic acids related to human health and how they interact with drugs and naturally occurring molecules that modulate their activity.


Collapse sponsor award id
S10RR025062

Collapse Time 
Collapse start date
2008-07-01
Collapse end date
2010-06-30

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