The Department of Biochemistry Metabolomics Facility
In a nutshell metabolomics is the profiling of all the small molecule metabolites found in a tissue, biofluid or cell. The approach has found applications in functional genomics, toxicology and safety assessment, the food industry, drug discovery and the pharmaceutical industry, the agrochemical industry and environmental toxicology. The Department of Biochemistry and more generally Cambridge has an excellent track record in metabolomics and has produced some of the seminal papers in the field. The department also boasts an impressive array of technology including the following dedicated instruments: a Bruker 500 MHz NMR spectrometer, two Thermo DSQ II GC-MS, a Thermo GC-FID, a Waters Quattro Premiere LC-MS and a Waters QToF Xevo LC-MS.
Because no one analytical technique can provide a complete coverage of the metabolome we have set out a number of either open (untargeted) or closed (targeted) approaches to profile metabolites for those wishing to carry out metabolomics. We are happy to discuss what is the most suitable method to use for your metabolomics experiments. We can also offer advice or carry out data processing of the subsequent data.
If you are interested in having samples analyzed please feel to discuss this with either:
1. High resolution 1H NMR spectroscopy of aqueous tissue extracts or biofluids to provide a metabolic fingerprint or footprint of the extract. Unassigned metabolites will be identified by standard 2D NMR spectroscopy and statistical approaches such as STOCSY followed by standard additions where possible. Other structure elucidation tools may also be used to help metabolite where appropriate. For tissue extracts, spectra could also be processed using specialised software (NMR suite Version 5.1, Chenomx, Edmonton, Canada) for peak fitting and semi quantification. Consumable cost: £25.50 per sample.
2. GC-FID analysis of total fatty acids. Fatty acids will be definitively identified by retention time matching with known standards and where necessary the acquisition of mass spectra using GC-MS and matching with compounds in the NIST library. Consumable cost: £24.00 per sample.
3. GC-MS analysis of the aqueous extract of tissues, cells or biofluids using a MSTFA derivatisation procedure as detailed in Atherton et al., 2006. Metabolites will be identified by mass spectral matching with the NIST library and other publically available resources e.g. the Fiehn lab database. Consumable cost: £27.00 per sample.
4. UPLC LC-MS analysis of lipids from tissue and serum extracts to identify bulk lipid changes as detailed in Roberts et al., 2008. This assay will use a UPLC QTOF Ultima and allows the analysis of intact lipids directly. Lipid identification will be carried out by MS/MS fragmentation and reference to libraries such as where possible. Consumable costs: £36.00 per sample.
5. Targeted analysis of aqueous metabolites using the triple quadrupole mass spectrometer. This will allow the quantitative analysis of over 65 aqueous metabolites and in particular amino acids and related compounds. The advantage of this approach is the complete assignment of the metabolites contributing to the Mass Spectral profile and absolute quantification. Consumable costs: £25.00 per sample. kit with a LC-MS
Consultant time to either prepare samples or process data: £40.00/hr. For large numbers of samples please discuss.
Tissue extracts can be prepared at a cost of £12 per sample.
Support time for further data processing: £37.50/hr. Arranged by discussion.
Costs to Industrial Clients:
Note: You must agree to our for Processing Samples - this assures you of confidentiality; no further agreements are necessary or will be entered into.
Charges as detailed above +50%+VAT.
1. Atherton HJ, Bailey NJ, Zhang W, Taylor J, Major H, Shockcor J, Clarke K, Griffin JL (2006) A combined 1H-NMR spectroscopy- and mass spectrometry-based metabolomic study of the PPAR-alpha null mutant mouse defines profound systemic changes in metabolism linked to the metabolic syndrome. Physiol Genomics. 2006 Oct 11;27(2):178-86.
2. Roberts, Lee D.; McCombie, Gregor; Titman, Christopher M.; Griffin, Julian L. (2008) A matter of fat: An introduction to lipidomic profiling methods. Journal of Chromatography, B: Analytical Technologies in the Biomedical and Life Sciences , 871(2), 174-181.