The following services are offered by the NERC Metabolomics Facility.
Before you apply to use the facility, we will discuss with you how best to design your metabolomics experiment. Please see "How to apply for services".
B. Sample Extraction
Extraction of metabolites from biological matrices.
This will be routinely carried out by visiting scientists after training provided by us. Each sample will be either extracted to provide a polar and non-polar fraction, based on the method by Bligh and Dyer, or by monophasic extraction.
C. Mass Spectrometry
We currently offer routinely:
- Nontargeted nanospray direct infusion MS (DIMS), using our "SIM-stitching" method, of polar and lipid metabolites in positive and negative ion modes, usually on a Thermo Q Exactive mass spectrometer.
- Nontargeted reversed-phase UHPLC MS using a Dionex U3000 RSLC system and the same mass spectrometer as above.
- Nontargeted Lipidomics using C30 columns on the same system.
- Targeted LC-MS./MS especially using MRM scanning on a Thermo TSQ Vantage.
Specific metabolites can be characterized with high mass accuracy (using internal standards), e.g. on a Bruker SolarX, and MSn fragmentation.
Other types of analysis are available, but might require additional development.
D. Nuclear Magnetic Resonance Spectroscopy
We currently offer 1-D and 2-D J-resolved NMR spectroscopy of polar metabolites on a Bruker Ultrashield 600 Plus, equipped with a cryoprobe.
Metabolite identification and quantification from 1-D NMR spectra can be conducted using the Chenomx NMR Suite. Visitors can conduct their own analyses in Birmingham, after training by NBAF-B staff. Structural elucidation using a range of further 2-D NMR techniques can be performed where required.
E. Data Analysis
Raw spectral data will be processed using mostly custom-written software. Subsequent data analysis will comprise of univariate and/or multivariate methods, and an initial output will be assembled from the statistical results and MS & NMR signal annotations (e.g. MIPack or Chenomx results). Further processing, statistical interrogations and identification can be undertaken following discussions of these initial results.
Visitors are welcome conduct their own statistical analyses, after training by NBAF-B staff. Our goal is to enable publication of the results.