Chemoselective Metabolomics via a Modular Reactivity-Encoding Platform

Area of Science:

• Biochemistry
• Analytical Chemistry
• Metabolomics

Background:

• Metabolic reprogramming is crucial for understanding diseases.
• Conventional liquid chromatography-mass spectrometry (LC-MS) struggles to detect all metabolites due to ionization and interference issues.
• This leads to underrepresentation of certain metabolite classes.

Purpose of the Study:

• To develop a chemoselective metabolomics strategy for functional group-resolved analysis.
• To overcome limitations of non-discriminatory metabolomics workflows.
• To enable comprehensive interrogation of the metabolome.

Main Methods:

• Developed a modular reactivity-encoding platform (MREP) using four alkyne-tagged probes.
• Probes selectively derivatize carboxyl, carbonyl, amine, and thiol groups.
• Encoded metabolites captured on a resin via azide-alkyne cycloaddition, removing interference and adding a reporter.

Main Results:

• Achieved high reaction orthogonality, near-quantitative conversion, and robust quantification.
• Triazole derivatives showed enhanced ionization and a universal reporter-ion for classification.
• Analyzed mouse serum and liver, identifying 7,208 features and 1,573 metabolites across four functional groups.

Conclusions:

• The MREP framework enables reactivity-resolved interrogation of complex small-molecule systems.
• This strategy significantly expands the detectable chemical space in metabolomics.
• Provides a versatile platform for detailed submetabolome classification and reconstruction.