4th April 2008 at 09:15 in the Benzon Auditorium

The Faculty of Pharmaceutical Sciences, Universitetsparken 2, 2100 Copenhagen

Bruker BioSpin GmbH, Silberstreifen, D-76287 Rheinstetten

09:15 - 10:15

NMR-based hyphenation in metabonomics

10:15 - 10:30

Coffee break

10:30 - 11:15

NMR system solutions for chemistry and biology

The lectures are an Drug Research Academy event organised by Jerzy Jaroszewski, e-mail jj(at)farma.ku.dk), C. Cornett, N. Nyberg and D. Stærk, Faculty of Pharmaceutical Sciences, University of Copenhagen.

The participation is free of charge and is open for attendance by all interested parties. No registration is required.

Abstract

NMR-based Hyphenation in Metabolomics

NMR is a major analytical technique used in Metabolomics. It can be be applied in high throughput screening using flow injection NMR. To allow rapid input a preparation robot has to be integrated, running under full automation. The second analytical method used regularely is mass spectroscopy. Optimum correspondance between NMR and MS can be achieved running samples n a synchronized mode in an integrated set up also including preparation. Such a system produces for example high quality heteroanalytical covariance analysis. Due to the lower sensitivity of NMR, many metabolites in a biofluid or extract will not be visible directly. A method to improve visibility in NMR is the combination with SPE to achieve a so-called SPE-NMR, where still mixtures are investigated; however a reproducable prefractionation and enrichment for example due to polarity can be achieved. Spectra obtained after elution can be investigated again with statistical methods and match algorythms. In addition the information from a LC-MS run can be available at the same time. A third method to apply NMR in Metabolomics involves isolation of new biomarkers or drug metabolites, this can be achieved running LC-NMR/MS, preferably combined with post column solid phase extraction and a cryogenic NMR probe. Such it is possible to investigate very low concentration metabolites with NMR and get important structural information. Examples are demonstrated for all applications mentioned.

NMR system solutions for chemistry and biology

Metabonomics has rapidly gained importance over the last years with NMR being the major analysis technology used. Besides NMR, mass spectroscopy, especially in combination with chromatography, is a preferred tool. NMR and MS are able to supplement each other in the information obtained.  NMR has its strength in the quantitative analysis of small molecules and can give an overview of changes in a biological sample without prior knowledge. MS is better suited for the analysis of large molecules in a mixture and of compounds in smallest concentrations. Having the technology of the ultrashielded magnets, it is now possible to combine flow-injection NMR and mass spectroscopy to a total analysis system with minimized space requirements and controlled by single software.  Such a system can also be used for sample preparation in an integrated manner. The instrument combination is described and examples are given. Software is also introduced that allows automatic evaluation of the spectra acquired by NMR and MS. The instrument setup can be used integrated or isolated, given outmost flexibility. Besides pharmaceutical samples, it can be used for clinical and food analysis applications as well as high throughput screening in parallel synthesis or combinatorial chemistry. Since optimized probehead technology is one of the prerequisites for successful Metabonomics work, they are described in more detail as part of the lecture.