Metabolite fingerprinting is a comparative high-throughput analysis with the intention to identify metabolite markers for genetic or environmental disturbances. For this the metabolic state of an organism under different experimental conditions is analyzed and compared (wild type vs. mutant or treated vs. non-treated organism). All metabolites possible are extracted and analyzed by Ultra Performance Liquid Chromatography coupled to a Time-of-Flight Mass Spectrometer (UPLC ESI-TOF-MS). Features which are showing intensity profiles of interest (e.g. higher amounts in treated organisms), are further analyzed and identified. The high coverage of almost all metabolites by extraction and analysis as well as the generation of accurate mass information by high resolution MS are a prerequisites to meet the aims of the non-targeted approach.
The metabolite fingerprinting analysis can be divided into five major steps:
1) Extraction of the metabolites from the biological samples.
2) Data acquisition by UPLC ESI-TOF-MS.
3) Peak picking and peak alignment for the generation of comprehensive data matrices.
4) Data processing and identification of metabolite markers (supported by the software tool MarVis)
5) Verification of the tentative identities of the metabolite markers by MS/MS or comparable methods.
Workflow: Data analysis by MarVis