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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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