Metabolomics Services
Our primary focus is on discovery (non-targeted) based metabolic profiling of biological fluids using liquid and/or gas chromatography coupled with mass spectrometry. The following illustrates the standard workflow that we utilize in a profiling experiment (GC- or LC-MS).
Step 1:
Small molecule metabolites are extracted from all means of biological fluids (plasma, serum, urine, etc.) to plant extracts and secreted fluids. |
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Step 2:
Separation of metabolites is performed using ultra performance liquid chromatography (UPLC, pictured at right) or gas chromatography.
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Step 3:
UPLC-MS: Detection of the metabololites is performed using a quadrupole time-of-flight mass spectrometer (Q-TOF). This instrument is capable of high mass accuracy measurements that can be used to calculate probable molecular formulas.
GC-MS: Detection is performed using an quadrupole mass analyzer.
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Step 4:
Alignment of the datasets by retention time and mass is performed using the Waters Marker Lynx software. This generates an aligned data matrix suitable for further processing with external software.
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Step 5:
Multivariate statistics are performed on the aligned data sets using the Umetrics SIMCA-P+ software. This type of analysis will illustrate the
maximum statistical separation between sample groups of interest.
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Step 6:
The "s-plot" is generated in the SIMCA-P+ software which aides in the process of generating a list of molecules are most heavily contributing to the separation between two of the groups.
These molecules of interest can then be further investigated:
• Molecular formula prediction
• Fragmentation of molecules (MS/MS)
• Comparison to standards
• Query existing databases
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