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Abstract No.: MoP-198
Session: Metabolomics, Metabonomics
Presentation date: Mon, Aug 28, 2006
Presentation time: 14:30 – 16:00

Evaluation and Application of GC-TOF-MS and Comprehensive GCxGC-TOF-MS to the Study of Complex Mammalian Metabolomes

Eva Zelena1, Warwick B. Dunn1, Jason J. Ashworth1, Rebecca Williams1, Maureen Cameron1, Project Consortium HUSERMET2, Douglas B. Kell1

1 The University of Manchester, Manchester, United Kingdom
2 http//www.metabolomics.co.uk/, Manchester, United Kingdom

Correspondence address: Eva Zelena, The University of Manchester, School of Chemistry, Faraday Building, Sackville Street, Manchester, M60 1QD United Kingdom.

Keywords: Mass Spectrometry, Gas Chromatography; Mass Spectrometry, Time of Flight; Metabolic Profiling; Metabolism, Metabolites.

Novel aspect: Development and application of comprehensive GCxGC-TOF-MS to metabolomic study of mammalian systems.

 

The metabolome is defined as the total quantitative collection of low molecular weight compounds (metabolites) present in cell or organism which participate in metabolic reactions required for growth, maintenance and normal function.1,2 The study of the metabolome (metabolomics) is an expanding scientific field employed in a variety of topics. Being ‘downstream’ of the genome/transcriptome/proteome, the metabolome represents the functional level of the cell and is the optimal stage for understanding and predicting genetic and phenotypic changes in the biology of complex biological systems. Mammalian systems have been shown to be the most complex biological systems studied and the number of metabolites are conservatively expected to number in the thousands.

By definition “true” metabolomics is described as the non-biased quantification of all metabolites within a biological system. However, because of the large chemical and physical complexity of metabolomes and the large dynamic concentration ranges observed this is not analytically achievable.3 Metabolic profiling is commonly employed to study a wide range of metabolites of different chemical and physical properties within a biological system. An array of analytical tools are available for studying the metabolome.3 GC-TOF-MS and comprehensive GCxGC-TOF-MS are currently being investigated as metabolic profiling tools as they provide excellent chromatographic separation, high sensitivity and the ability to identify metabolites via mass spectral fragmentation patterns. The optimisation of analytical instrumentation is an important requirement to maximise the biological information obtained from complex metabolomes. As has been undertaken for GC-TOF-MS4 we have implemented an entirely automated (closed-loop) strategy for multi-objective (peak number, S/N, analysis time) optimisation of comprehensive GCxGC-TOF-MS instrumentation.

The assessment of GC-TOF-MS and comprehensive GCxGC-TOF-MS for the study of mammalian biofluids (serum/plasma and urine) has been performed. Discussions on automated closed-loop optimisation, improved chromatographic resolution, sensitivity, mass spectral purity and metabolite identification will be provided. The study of storage temperature, protein precipitation procedures will be discussed as will the investigation of clinical disease biomarkers.

1. S. G. Oliver, M. K. Winson, D. B. Kell and F. Baganz, Trends Biotechnol. 16, 373 (1998).
2. C. W. W. Beecher in Metabolic Profiling: Its Role in Biomarker Discovery and Gene Function Analysis, Ed. by G.G. Harrigan and R. Goodacre, Kluwer Academic Publishers, London, p. 311 (2003).
3. W. B. Dunn, N. J. Bailey and H. E. Johnson, Analyst 130, 606 (2005).
4. S. O’Hagan, W. B. Dunn, M. Brown, J. D. Knowles and D. B. Kell, Anal. Chem. 77, 290 (2005).