We have done the analysis for a pre-clinical study and we have observed additional peaks in the chromatograms. We think that these peaks are metabolites. The concentration of the additional peaks are very low (we expect it to be between 0.5-1 ng/mL). How can we find out what the structure is, or prove it is a metabolite or not? I wish to get global overview how to solve this matter.
Thyoe of solvent depends on type of metaboloites,. Is it polar or non polar also depends on physical and chemical properties of metabolites
That is probably going to be a challenging endavour! I'm not an expert in the field of metabolite ID, but I don't think you can deduce the identity of a component from the chromatogram (retention time) only. Unless you can predict the main metabolites of your compound of interest, have these synthesized and determine the retention times of these in your chromatographic method.
Otherwise, use LC-MS/MS to get an accurate mass (i.e. elemental composition) of the suspected metabolite, as well as a fragmentation spectrum, this will aid in deciphering the identity and perhaps structure of the molecule. If you compare the MS data to that obtained for the starting compound, you can look for similarities in the MS/MS data, which could suggest that you're indeed looking at metabolites of a certain component.
Hope this helps..
Mr. Asali,
Please, consider the following discussion and the reference section, therein.
[https://www.researchgate.net/post/How_to_analyze_two_different_time_point_metabolomics_data]
High-resolution accurate mass spectrometry analysis should be performed.
If you define clearly what you perceive as "metabolite" it will become possible to provide some answers
Use the Liquid Chromatography coupled to High-Resolution Mass Spectrometry and do the MS/MS analysis for identifying the unknown metabolites.
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