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Miniaturized Two-Dimensional Heart Cutting for LC-MS-Based Metabolomics.
Carla Orlandi1,2, Carine Jacques3, Hélène Duplan3
1Toxalim (Research Centre in Food Toxicology), INRAE UMR 1331, Paul Sabatier University (UPS), ENVT, INP-Purpan, Toulouse 31062, France.
Analytical Chemistry
|January 30, 2023
Summary
This study introduces a novel 2D-LC-MS workflow that integrates hydrophilic interaction liquid chromatography (HILIC) and reversed-phase (RP) separations. This method efficiently analyzes polar and nonpolar metabolites from a single injection, reducing sample consumption and analysis time.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Metabolomics
Background:
- Liquid chromatography-mass spectrometry (LC-MS) metabolomics often requires separate hydrophilic interaction liquid chromatography (HILIC) and reversed-phase (RP) workflows.
- These separate workflows lead to high sample consumption and extended analysis times.
- A need exists for integrated methods to improve efficiency in metabolomic analyses.
Purpose of the Study:
- To develop and validate an integrated 2D-LC-MS workflow combining HILIC and RP separations.
- To enable comprehensive metabolome coverage with reduced sample volume and analysis time.
- To demonstrate the workflow's utility in analyzing complex biological samples like human plasma.
Main Methods:
- Development of miniaturized, independent 1D HILIC and RP methods using 1 mm inner diameter columns.
- Integration of HILIC and RP methods into a 2D micro LC-MS system using heart-cutting trapping.
- Optimization of chromatographic and ionization conditions, including trapping parameters.
- Application of the workflow to human plasma standard reference material SRM 1950.
Main Results:
- Successful integration of HILIC and RP separations into a single injection workflow.
- Detection of hundreds of metabolites in human plasma, including both polar and nonpolar compounds.
- Significant reduction in sample injection volume to 0.5 μL.
- Minimized loss of certain metabolites compared to separate workflows.
- Reduced overall analysis time due to column equilibration during elution.
Conclusions:
- The developed integrated 2D-LC-MS workflow offers a more efficient approach to metabolomics.
- This method enhances the coverage of diverse metabolites while minimizing sample requirements.
- The workflow is suitable for analyzing complex biological matrices, improving metabolomic study efficiency.
