The Mason Metabolomics Facility
Nonvolatile Metabolites
Nonvolatile metabolites are isolated from biological samples by a combination of mechanical homogenization (if necessary), ultrafiltration (to remove particulates and macromolecules), and organic extraction. The organic and/or aqueous fractions may be characterized. Solid phase extraction (SPE) may also be employed, to concentrate low abundance analytes. Targeted metabolomics analyses are performed using appropriate standards and our Agilent LCMS or Agilent LC-QToF (MS/MS) equipped with a reverse-phase (C5 or C18 for the organic soluble metabolites) or HILIC (for the aqueous soluble metabolites) chromatography column. The ion source selected (ESI, APPI, or APCI) is determined by compatibility with the chromatography solvent and analytes under investigation. Analysis is typically performed at either pH 3 or pH 9 (or both, where appropriate), to affect the ionization state of the analytes, and thereby maximize the chromatographic content. Global (aka discovery) metabolomics analyses are performed using our Agilent LC-QToF (MS/MS). Sample preparation is as described above, and the chromatography performed using reverse-phase and/or normal-phase columns.
An Example Nonvolatile Study:
Tissue metabolomes are clearly distinguishable (cluster) by tissue type in a PCA of pig organs.
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