Chris L. Stumpf

Metabonomic analysis of mouse urine by liquid-chromatography-time of flight mass spectrometry (LC-TOFMS): detection of strain, diurnal and gender differences

The application of HPLC-MS combined with principal components analysis (PCA) to the metabonomic analysis of mouse urine is demonstrated. Urine samples from three strains of mouse were analysed by gradient HPLC-MS combined with positive and negative electrospray time-of-flight mass spectrometry. Analysis of the resulting data using PCA enabled the samples to be discriminated between on the basis of gender, strain and diurnal variation. These preliminary results suggest that HPLC-MS-based approaches may have a useful role in metabonomic analysis that complements existing approaches.

A multi-analytical platform approach to the metabonomic analysis of plasma from normal and zucker (fa/fa) obese rats

Plasma obtained from 20 week old normal Wistar-derived and Zucker (fa/fa) rats was analysed using a number of different analytical methodologies to obtain global metabolite profiles as part of metabonomic investigations of animal models of diabetes. Samples were analysed without sample pre-treatment using 1H NMR spectroscopy, after acetonitrile solvent protein precipitation by ultra-performance liquid chromatography-MS (UPLC-MS) and after acetonitrile protein precipitation and derivatisation for capillary gas chromatography-MS (GC-MS). Subsequent data analysis using principal components analysis revealed that all three analytical platforms readily detected differences between the plasma metabolite profiles of the two strains of rat. There was only limited overlap between the metabolites detected by the different methodologies and the combination of all three methods of metabolite profiling therefore provided a much more comprehensive profile than would have been provided by their use individually.