Methods of chemoselective derivation of multiple classes of metabolites
Chemoselective derivatization of biological amines, carboxylic acids, aldehydes or ketones are employed in methods to detect a plurality of components, or members of a component, such as metabolites, that vary in molecular structure. The methods of the invention can be employed in aqueous and nonaqueous conditions.
1. A method of characterizing metabolites in a biological sample, comprising the steps of:
a) combining at least a portion of a biological sample, the biological sample including at least one metabolite that is, or includes,
i) an amino acid or an amine,
ii) an aldehyde or a ketone, or
iii) a carboxylic acid or an alcohol,
with an ultraviolet-active isocyanide and Ugi reactants that consist essentially of two of i), ii) and iii) apart from the metabolite, thereby derivatizing at least a portion of that metabolite; and
b) characterizing the derivatized metabolite of the biological sample.
2. The method of claim 1 , wherein the metabolite is a first metabolite, and wherein the biological sample includes a second metabolite that is, or includes,
i) an amino acid or an amine,
ii) an aldehyde or a ketone, or
iii) a carboxylic acid or an alcohol,
that is not any of i), ii) or iii) of the first metabolite, and further including the steps of
a) combining at least a portion of the biological sample with an ultraviolet-active isocyanide and Ugi reactants that consist essentially of two of i), ii) and iii) apart from the second metabolite, thereby derivatizing at least a portion of the second metabolite; and
b) characterizing the derivatized second metabolite.
3. The method of claim 1 , wherein the metabolite is selected from the group consisting of polyamines, fatty acids, neurotransmitters, hormones, α-amino acids, intermediates in central metabolism, and co-enzymes.
4. The method of claim 1 , wherein the metabolite includes an amine.
5. The method of claim 1 , wherein the metabolite includes an aldehyde.
6. The method of claim 1 , wherein the metabolite includes a ketone.
7. The method of claim 1 , wherein the metabolite includes a carboxylic acid.
8. The method of claim 1 , wherein the metabolite includes an α-amino acid.
9. The method of claim 1 , wherein the biological sample includes a plurality of chemically distinct metabolites sharing a common functional group of,
i) an amino acid or an amine,
ii) an aldehyde or a ketone, or
iii) a carboxylic acid or an alcohol.
10. The method of claim 9 , wherein the step of characterizing the derivatized metabolites includes separating at least a portion of the derivatized metabolites from each other.
11. The method of claim 10 , wherein the derivatized metabolites are separated from each other by liquid chromatography.
12. The method of claim 11 , wherein the derivatized metabolites are characterized, at least in part, by mass spectroscopy.