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Extraction of Aqueous Metabolites from Cultured Adherent Cells for Metabolomic Analysis by Capillary Electrophoresis-Mass Spectrometry
Published on: June 9, 2019
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Characterization of rapid extraction protocols for high-throughput metabolomics.
Sarah Gehrke1, Julie A Reisz1, Travis Nemkov1
1Department of Biochemistry and Molecular Genetics, University of Colorado Denver - Anschutz Medical Campus, 12801 East 17th Ave, Aurora, CO, 80045, USA.
Rapid Communications in Mass Spectrometry : RCM
|June 7, 2017
Summary
Rapid hydrophilic compound extraction from blood, plasma, and red blood cells is feasible in ~4 minutes. This fast method enables quantitative metabolomics for clinical applications, overcoming previous time limitations.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Clinical Diagnostics
Background:
- High-throughput metabolomics advances research and personalized medicine.
- Current methods struggle with rapid extraction for time-sensitive clinical needs, like emergency departments.
Purpose of the Study:
- To assess fast liquid-liquid extraction for hydrophilic compounds.
- To enable quantitative metabolomics in clinical settings.
Main Methods:
- Ultra-high-pressure liquid chromatography/mass spectrometry (UHPLC/MS).
- Fast liquid-liquid extraction of hydrophilic compounds.
- Use of stable-isotope-labeled internal standards for quantification.
- Analysis of human whole blood, plasma, and red blood cells.
Main Results:
- Extraction yields were unaffected by vortexing time (4-60 min) for most hydrophilic metabolites.
- High quantitative accuracy (CV <20%) was achieved for most compounds.
- Recoveries exceeded 80% for most metabolites, with 50-75% for basic amino acids and polyamines.
- Plasma and red blood cell metabolomes showed similarities.
Conclusions:
- Rapid (~4 min) hydrophilic compound extraction is effective.
- This method is viable and potentially automatable for quantitative analysis.
- Supports research and clinical applications in metabolomics.
