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A Strategy for Sensitive, Large Scale Quantitative Metabolomics
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High-throughput and multiplexed LC/MS/MRM method for targeted metabolomics.

Ru Wei1, Guodong Li, Albert B Seymour

  • 1Metabolomics/Proteomics, Applied Quantitative Genotherapeutics, Pfizer Inc. 620 Memorial Drive, Cambridge, Massachusetts 02139, USA. ru.wei@pfizer.com

Analytical Chemistry
|June 8, 2010
PubMed
Summary
This summary is machine-generated.

This study presents a new high-throughput method for target-based metabolomics, enabling the analysis of 205 endogenous metabolites in just 10 minutes. This sensitive and reproducible technique enhances the study of metabolic changes in various biological and disease states.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Systems Biology

Background:

  • Target-based metabolomics is crucial for understanding metabolic alterations in diseases and responses to stimuli.
  • Analyzing numerous endogenous metabolites simultaneously is challenging due to their diverse chemical properties.

Purpose of the Study:

  • To develop a high-throughput, sensitive, and reproducible method for target-based metabolomics.
  • To enable the simultaneous analysis of hundreds of endogenous metabolites from complex biological samples.

Main Methods:

  • Utilized a liquid chromatography-tandem mass spectrometry (LC/MS/MRM) system.
  • Combined optimized separation conditions, ionization polarities, and multiple reaction monitoring (MRM) acquisition.
  • Sequentially analyzed metabolites in three subgroups: amino acids, sugars/nucleic acids, and organic acids.

Main Results:

  • Successfully analyzed 205 endogenous metabolites in 10 minutes.
  • Achieved low picogram sensitivity for over half of the analyzed metabolites.
  • Demonstrated 3-4 orders of linearity and assay coefficients of variation <15% for approximately 80% of metabolites.

Conclusions:

  • Established a multiplex LC/MS/MRM method for quantitative profiling of hundreds of known metabolites.
  • The developed methodology is broadly applicable and expandable for analyzing endogenous and drug metabolites.
  • This method provides a robust tool for advancing metabolomics research in diverse applications.