The equipment requested in this application is a 700 MHz microcoil NMR probe with a sample handling system capable of loading samples from 96-well plates under unattended automation. Additionally, we are requesting an electrophoretic sample preparation system, Dynamic Field Gradient Focusing (DFGF), capable of performing most capillary electrophoresis (CE) methods on a preparative scale appropriate to the micro-NMR probe and transferring purified bands into the automated loading system. Nuclear Magnetic Resonance (NMR) spectroscopy is highly informative for chemical analysis, but its sensitivity is limited relative to other methods such as mass spectrometry (MS). Small-volume solenoidal NMR detectors (microcoil probes) offer gains in sensitivity comparable to using a cryoprobe at higher field, but are much more cost-effective for mass-limited samples such as those encountered in LC-MS analysis, and can be exchanged with other probes for different applications. Preliminary data obtained with a loaned microcoil probe on a 500 MHz NMR spectrometer is presented to establish its capabilities for trace analysis in our major applications. The automation system provides effective LC-MS-NMR capability when used in combination with a """"""""nanosplitter"""""""" LC-MS interface available at the host institute (Barnett Institute of Chemical and Biological Analysis);the nanosplitter provides high LC-MS performance while collecting 98% of LC flow into fractions for subsequent NMR analysis. Similarly, the DFGF system can extend CE or CE- MS analysis to CE-NMR. The major applications are natural product discovery, identification of DNA adducts, glycan analysis for glycoprotein cancer biomarkers and for biosimilar protein therapeutics, and metabolite profiling for toxicology in drug development. Other applications include screening anticancer agents, characterizing the oral microbiome, and metabolomics.
The proposed instrumentation provides rapid and accurate chemical identification to enhance a number of active programs, with goals to (i) develop new medicines, from both natural and synthetic sources, and (ii) to detect changes in the body chemistry associated with disease, reveal its underlying causes, and detect it earlier, thereby providing more successful treatments.
| Kautz, Roger; Wang, Poguang; Giese, Roger W (2013) Nuclear magnetic resonance at the picomole level of a DNA adduct. Chem Res Toxicol 26:1424-9 |
