Pilot study for targeted analyses

Details

Last Updated: Friday, 04 September 2020 14:21

• The method must have a simple (preferably one step) and defined extraction procedure which can be used to generate an extract from the matrix suitable for metabolite analysis by mass spectrometry.
• The metabolites of interest must be stable in the extract until analysis by GC-MS or LC-MS.
• The metabolites must have a unique defined retention time (RT) on a chromatographic column, mass signal (molecular ion or eq.) in a full scan analysis and/or unique transition from a tandem mass analysis (e.g. MRM).
• Internal standards with similar chemical structure and chemical behavior must be available for quantification of the target metabolites.
• There should be a linear response at least in a magnitude order of 10³ between the instrumental signal (MS detector) and each concentration of the target metabolites.
• The response of a metabolite must increase when the matrix extract is spiked with its corresponding standard.
• It must be possible to detect the metabolites within the linear concentration range

• A list of relevant references from the scientific literature
• Definition of the utilized matrix (e.g. cell lines or plasma)
• A protocol for sample preparation and extraction of the metabolites from the defined matrix
• A protocol describing instrumental parameters and the analytical procedure
• A chromatogram of standards and internal standards, both in organic solvent and in spiked matrix extract (high and low concentrations)
• Estimated detection limits of target metabolite standards
• Data showing the linear response of the target metabolites
• Data confirming that the metabolites are stable in the extract until the time of analysis
• Results showing the signal of the target metabolites in the defined matrix when they are spiked by a defined concentration of metabolite standards