Nanoparticle microarray for high-throughput microbiome metabolomics using matrix-assisted laser desorption ionization...

Area of Science:

• Metabolomics
• Microbiome Analysis
• Mass Spectrometry

Background:

• The gut microbiome plays a crucial role in host health.
• Understanding metabolomic profiles is key to assessing microbiome function.
• High-throughput methods are needed for comprehensive microbiome metabolomics.

Purpose of the Study:

• To develop a high-throughput matrix-assisted laser desorption/ionization mass spectrometry (MALDI)-MS platform for microbiome metabolomics.
• To optimize nanoparticle and organic matrix combinations for enhanced metabolite detection.
• To apply the platform to analyze metabolomic changes in turkey gut samples under different antibiotic treatments.

Main Methods:

• Developed a pre-fabricated microarray platform utilizing nanoparticles and organic matrices for MALDI-MS.
• Screened various inorganic and metal nanoparticles (NPs) and organic matrices for optimal performance.
• Analyzed turkey intestinal samples using the optimized MALDI-MS platform, coupled with LC-MS/MS for metabolite identification.

Main Results:

• Identified an optimal matrix set including specific NPs (Fe3O4, Au, Cu, Ag) and an organic matrix (α-cyano-4-hydroycinnamic acid).
• Reproducibly detected over two thousand unique metabolite features across turkey gut samples.
• Chemically identified 56 metabolites using MALDI-MS/MS and LC-MS/MS, with significant metabolomic variations observed based on age and antibiotic treatment.

Conclusions:

• The developed MALDI-MS platform offers a high-throughput and reproducible approach for microbiome metabolomics.
• The study successfully demonstrated the platform's utility in revealing age- and treatment-dependent metabolic variations in the turkey gut microbiome.
• This method provides a powerful tool for investigating the impact of diet and antibiotics on gut microbial metabolism.