High-Frequency Microfluidic Fractionation for Compound-Resolved Bioactivity-Based Metabolomics

Area of Science:

• Analytical Chemistry
• Metabolomics
• Drug Discovery

Background:

• Specialized metabolites are vital drug candidates, but their identification from extracts is slow.
• Nontargeted metabolomics faces bottlenecks in functional assessment, hindering biological understanding.
• Urgent need for new drugs, especially antibiotics, necessitates faster discovery methods.

Purpose of the Study:

• To develop a compound-resolved bioactivity-based metabolomics workflow.
• To accelerate the identification of active agents from complex metabolite samples.
• To address the bottleneck in functional assessment within metabolomics.

Main Methods:

• Combined nontargeted liquid chromatography tandem mass spectrometry (LC-MS/MS) with high-frequency fractionation.
• Utilized microfluidic paper-analytical devices (μPADs) for parallel spotting during MS/MS acquisition.
• Employed luminescent bioreporter strains to detect cellular stress and correlate signals with MS data.

Main Results:

• Demonstrated high sensitivity (down to 1 ng/spot) for antimicrobial compound detection.
• Successfully identified multiple antimicrobial compounds from crude extracts of *Saccharopolyspora erythraea*.
• Highlighted the workflow's practicality and high-throughput capability for drug discovery.

Conclusions:

• The developed workflow enables rapid, compound-resolved bioactivity-based metabolomics.
• This approach significantly speeds up the identification of potential drug candidates from complex samples.
• The method advances systematic functional assessment in metabolomics and aids antibiotic discovery.