Code: WOE	Time Slot/Poster Number: 4:00-4:25	Session: Hyphenated Techniques
Advanced LC/microcoil NMR methods for structure determination and metabolite profiling
Daniel Raftery; Ravi KC; Emmanuel Appiah-Amponsah; Kwadwo Owusu-Sarfo; Tao Ye; GA Nagana Gowda
Purdue University, West Lafayette, IN
Abstract
The analysis of complex samples such as biofluids poses a number of analytical challenges. While the combination of NMR with multivariate statistics provides a successful approach to analyze high concentration species for metabolite profiling, challenges still exist. In order to delve more deeply into the complex samples and analyze lower concentration species that may be more sensitive or specific to changes in biological states, more advanced methods are attractive. In this presentation we will discuss the combination of chromatography and/or fractionation with advanced microcoil methods to identify low concentration metabolites of sometimes unknown structures. This approach is very complementary to the more conventional methods of whole fluid analysis and the knowledge gained can be leveraged back for high throughput analysis.

Code: WOE	Time Slot/Poster Number: 4:25-4:40	Session: Hyphenated Techniques
Combining DEER and PRE Distance Restraints with Solution-State NMR Data for Structure Determination of Protein Complexes
Yunhuang Yang; Theresa A. Ramelot; Michael A. Kennedy
Miami University, Oxford, OH
Abstract
There is increasing integration of complementary methodologies for structure determination of multi-protein complexes. One approach is to supplement conventional solution-state NMR data with intermediate and long-range distances (10-70Å) from paramagnetic relaxation enhancements (PRE) and EPR-based Double Electron Electron Relaxation (DEER) for structure validation, filtering candidate structures, or for de novo structure calculations. Here, we describe integration of PRE and DEER data with solution NMR data for structure determination of a 13.6 kD homodimer. Conventional DEER enabled inter-chain distance measurements out to 34Å and novel DEER experiments enabled direct measurement of inter-chain exchange kinetics. PRE and DEER distances validated NMR and X-ray structures and were sufficient to generate correct inter-chain packing in the absence of C13-edited/C12-filtered NOEs or RDCs.

Code: WOE	Time Slot/Poster Number: 4:40-5:05	Session: Hyphenated Techniques
Hyphenated Technologies - PET/MR
T. Adrian Carpenter1; Robert C Hawkes1; Tim D Fryer1; Richard E Ansorge2; Joseph W Stevick1; Stefan B Siegel3
1Wolfson Brain Imaging Centre, Cambridge, United Kingdom; 2Department of Physics, University of Cambridge, Cambridge, United Kingdom; 3Siemens Molecular Imaging, Preclinical Solutions, Knoxville, TN 37932
Abstract
PET/MR is an exciting hyphenated technology for molecular imaging. Prototype system for preclinical and clinical studies are now available. We describe the Cambridge preclinical system based on a 1T split magnet and a state of the art Siemens F120 PET scanner and contrast this with solid state Photodiode approaches. The performance characteristics of the system are discussed and preliminary simultaneous PET/MR images of a biological system will be presented.

Code: WOE	Time Slot/Poster Number: 5:05-5:20	Session: Hyphenated Techniques
A Superconducting Magnet for NMR-Mass Simultaneous Measurements
Kentaro Kominato
Japan Superconductor Technology, Inc., Kobe, Japan
Abstract
We developed a novel spectroscopic magnet. The new spectroscopy, which was suggested by Fuke, would enable to carry out NMR and Mass spectroscopy simultaneously. In this methodology nuclear magnetic resonance is observed as acceleration of the ions taking advantage of interaction between nuclear magnetic moment and magnetic field gradient. In order to realize the methodology, the magnet is required to provide two homogeneous fields of 12 and 4 tesla, respectively. The field gradient between these two regions must be high enough to accelerate ions. The coil structure is robust against high stress due to the asymmetric magnetic field. The magnet is shielded actively and has a horizontal access room temperature bore of 155 mm diameter.

Code: WOE	Time Slot/Poster Number: 5:20-5:45	Session: Hyphenated Techniques
Roles of the hyphenated HPLC-DAD/MS-SP E-NMR technique in metabonomics
Yulan Wang
Wuhan Institute Of Physics And Mathematics, Wuhan,
Abstract
Metabolite identification is a key step for metabonomics study. Hyphenation of high performance liquid chromatography-diode-array detector/mass spectrometry-solid phase extraction-NMR spectroscopy (HPLC-DAD/MS-SPE-NMR) represents the most important analytical techniques for the development of metabonomics. HPLC allows the separation of a metabolite from a mixture whereas SPE unit facilitates multiple trapping of unknown metabolites. There are two benefits: one is enrichment of a metabolite, which overcomes the drawback of low sensitivity of NMR spectroscopy, the other is avoidance to perform solvent suppression in NMR experiment. HPLC-DAD/MS-SPE-NMR hyphenation provides a fast method for structure elucidation of novel and trace metabolites in plant extracts and drug metabolites. Modification of this hyphenated technique, enabling multiple trappings of strong polar metabolites for biofluids analysis, needs further development.