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Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry UPLC-HRMS
Published on: May 20, 2013
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Chiral High-Performance Liquid Chromatography (HPLC) in Targeted Metabolomics
Xiaoqing Fu1,2, Michael Lämmerhofer3
1Pharmaceutical (Bio-)Analysis, Institute of Pharmaceutical Sciences, University of Tübingen, Tübingen, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|November 22, 2025
Summary
Stereoselective analysis is crucial for understanding biological processes. This study details targeted enantioselective liquid chromatography-tandem mass spectrometry methods for analyzing key metabolites like oxylipins and fatty acids.
Area of Science:
- Analytical Chemistry
- Metabolomics
- Lipidomics
Background:
- Stereoselectivity is vital in enzymatic reactions and understanding biological processes.
- Distinguishing stereoisomers, especially enantiomers, requires specialized separation techniques.
- Current methods like untargeted metabolomics have limitations in comprehensive enantiomer analysis.
Purpose of the Study:
- To present targeted enantioselective liquid chromatography (LC) assays for critical metabolite classes.
- To highlight the application of LC coupled with tandem mass spectrometry (MS) for stereoisomer analysis.
- To provide robust methodologies for enantioselective analysis in biological samples.
Main Methods:
- Utilized targeted enantioselective LC with chiral stationary phases.
- Employed triple quadrupole (QqQ) mass spectrometry for sensitive detection.
- Developed specific workflows for oxylipins, 3-hydroxy fatty acids, amino acids, and short branched-chain fatty acids.
Main Results:
- Successfully applied enantioselective LC-MS/MS assays to analyze diverse metabolite classes.
- Demonstrated applicability for samples from platelets and plasma.
- Established protocols for proteinogenic amino acid and short branched-chain fatty acid enantiomer separation.
Conclusions:
- Targeted enantioselective LC-tandem MS is a powerful technology for stereoisomer analysis in metabolomics.
- The presented methods offer broad metabolite class coverage with high specificity.
- These assays are essential for a complete understanding of metabolic states and biological pathways.
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