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Related Experiment Video

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Dual UHPLC-HRMS Metabolomics and Lipidomics and Automated Data Processing Workflow for Comprehensive High-Throughput

P Vangeenderhuysen1, J Van Arnhem1, B Pomian1

  • 1Laboratory of Integrative Metabolomics (LIMET), Department of Translational Physiology, Infectiology and Public Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.

Analytical Chemistry
|May 23, 2023
PubMed
Summary

Fecal metabolomics and lipidomics can now be performed faster using a novel dual extraction and UHPLC-HR-Q-Orbitrap-MS workflow. This method, coupled with the TaPEx algorithm, significantly reduces sample-to-result time for gut microbiome research.

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Area of Science:

  • Metabolomics and Lipidomics
  • Gut Microbiome Research
  • Analytical Chemistry

Background:

  • Feces is a valuable, non-invasive matrix for studying the gut microbiome-health axis.
  • High-throughput analysis is crucial for large cohort studies with limited sample availability.
  • Existing methods require optimization for speed, efficiency, and comprehensive molecular profiling.

Purpose of the Study:

  • To develop a high-throughput, dual fecal extraction workflow for simultaneous metabolome and lipidome analysis.
  • To validate a novel R-based targeted peak extraction (TaPEx) algorithm for automated data processing.
  • To reduce sample-to-result time in fecal microbiome analyses.

Main Methods:

  • A dual fecal extraction method combined with ultra-high performance liquid chromatography-high resolution-quadrupole-orbitrap-mass spectrometry (UHPLC-HR-Q-Orbitrap-MS).
  • Analysis of 836 in-house standards, detecting 360 metabolites and 132 lipids.
  • Optimization and benchmarking of the TaPEx algorithm against other software using cohort samples.

Main Results:

  • Successful targeted profiling with high repeatability (78% CV < 20%), reproducibility (82% CV < 20%), and linearity (81% R² > 0.9).
  • Detection of 15,319 features in untargeted fingerprinting (CV < 30%).
  • TaPEx algorithm outperformed untargeted approaches, achieving 81.3% compound detection.
  • A 60% reduction in sample-to-result time was achieved in cohort studies.

Conclusions:

  • The developed dual fecal extraction and UHPLC-HR-Q-Orbitrap-MS workflow enables efficient, high-throughput metabolome and lipidome analysis.
  • The automated TaPEx algorithm significantly improves targeted data processing efficiency.
  • This integrated approach accelerates gut microbiome research, facilitating large-scale cohort studies.