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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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Bioinert UHPLC system improves sensitivity and peak shapes for ionic metabolites
Ondřej Peterka1, Alena Langová1, Robert Jirásko1
1University of Pardubice, Faculty of Chemical Technology, Department of Analytical Chemistry, Studentská 573, 53210 Pardubice, Czech Republic.
Journal of Chromatography. A
|December 11, 2024
Summary
Bioinert materials in chromatography improve peak shape and sensitivity for analyzing ionic metabolites, especially those with phosphate groups. This advancement enhances untargeted metabolomic analysis in complex samples like human plasma.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Metabolomics
Background:
- Ionic compound analysis via liquid chromatography faces challenges due to analyte interactions with metal surfaces, causing poor peak shape and reduced sensitivity.
- Bioinert materials in chromatographic systems mitigate these unwanted interactions, improving analytical performance.
Purpose of the Study:
- To develop and optimize an ultrahigh-performance liquid chromatography (UHPLC) method coupled with high-resolution mass spectrometry for untargeted metabolomic analysis using bioinert components.
- To evaluate the impact of bioinert technology on the analysis of ionic metabolites, particularly those containing phosphate groups.
Main Methods:
- Utilized UHPLC with bioinert components connected to a high-resolution mass spectrometer.
- Developed and optimized the method using 81 metabolite standards.
- Assessed method performance through calibration curves, dynamic range, limit of detection, and linearity.
- Applied the optimized method to analyze NIST SRM 1950 human plasma.
Main Results:
- Bioinert technology significantly improved peak shapes and sensitivity compared to conventional systems, especially for phosphate-containing metabolites.
- Calibration curves demonstrated a wide dynamic range, low limits of detection, and excellent linearity (R² > 0.99).
- Analysis of human plasma identified 156 metabolites and polar lipids with high confidence using mass accuracy, MS/MS fragmentation, and chromatographic behavior.
Conclusions:
- The developed UHPLC-MS method using bioinert components effectively enhances the analysis of ionic metabolites, offering improved peak shape and sensitivity.
- This bioinert technology shows significant potential for the analysis of various ionic compounds, including those with phosphate groups, in complex biological matrices.
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