Metabolomics techniques allow the analysis of metabolites in biofluids and tissues giving metabolic profiles as the end product. Such profiles provide a unique fingerprint of the metabolic state of an organism and can be used to identify specific metabolic changes leading to the understanding of biochemical pathway, biomarker combinations, toxic effects and disease progression. NMR spectroscopy and mass spectrometry are the two most common analytical techniques used to specifically identify metabolites or generate metabolic profiles. NMR is mainly used to detect hydrogen atoms in metabolites. The NMR analysis detects all proton-containing small metabolites (<1500 Da) present in concentrations above ca. 5 microM. Thus, all hydrogen-containing molecules will give a proton NMR spectrum, as long as they are present in concentrations above the detection limit. Different molecules produce different NMR signals and NMR spectra of biological samples are therefore the superposition of the spectra of all metabolites in the sample. The relative intensities of these signals reflect the concentration of each metabolite. The datasets obtained are further analyzed using multivariate analysis to link the NMR data to biological knowledge. The main applications of metabolomics to humans are related to phenotyping, biomarker discovery, clinical studies (diagnosis disease and therapeutic efficacy), toxicology...
We offer assistance in defining the nature and dimension of the sample set in each metabolomic study to be performed, with the aim of ensuring sound and significant results from the very beginning. The services provided include: experimental design, sample preparation, NMR analysis and assistance in data processing and analysis. Hands-on training in the application of NMR-based metabonomics is also proposed.
- 600 MHz NMR spectrometer equipped with a 5 mm 1H/13C/15N/31P cryoprobe and a
sample changer allowing storage at + 6 °C and automatic measurement of biofluids and
- 600 MHz NMR spectrometer equipped with a 4 mm 1H/13C high resolution magic angle
spinning probe for studies of intact tissues
We also offer some complementary metabolomics analysis using mass spectrometry (LC-MS/MS)
We also perform non-metabolomics NMR analysis (structural analysis, interaction studies..)
|Input samples||Whole blood, Plasma, Serum, Fluids (urine, saliva, CSF, etc), Solid tissue, Tissue cells, Tissue extract|
|Sample formats||- Biofluids and tissue extracts are mixed with a deuterated phosphate buffer or saline solution and transferred into NMR tubes - Intact tissues (ca. 15 mg) are mixed with a small amount of D2O or saline solution (typically 20-50 microliters) and transferr|
|Analysis throughput||1 day / 10-100 samples||The time required for standard data collection by NMR is between 10 to 60 min depending on the type of sample and number of NMR experiments. A standard acquisition time for a simple one-dimensional proton NMR spectrum of urine is ca. 10 min. / After sample preparation, 120 urine samples, 48 serum/plasma samples or 72 extracts can be for example analyzed (ref. 1 in Protocols).|
|Analysis costs||Contact the manager||The cost depends on the type of sample to be analyzed (biofluid, extract, intact tissue). For samples such as urine and serum that require minimal preparation and for acquisition of a simple one-dimensional 1H NMR spectrum, the cost per sample is 300 SEK for academics. The cost also depends on the user/staff participation in the sample preparation and analysis: user operation after training, full service, and projects where the investigator collaborate with the platform staff in experimental design, experimentation, troubleshooting, and data analysis.|
- O. Beckonert et al., Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts, Nature Protocols, 2, 2692-2703, 2007
- O. Beckonert et al., HR MAS NMR for metabolic profiling of intact tissues
Nature Protocols, 5,1019-1032, 2010