A Comprehensive LC–MS Metabolomics Assay for Quantitative Analysis of Serum and Plasma Article Swipe
Lun Zhang
,
Jiamin Zheng
,
Mathew Johnson
,
Rupasri Mandal
,
M. D. Soler Cruz
,
Miriam Martínez‐Huélamo
,
Cristina Andrés‐Lacueva
,
David S. Wishart
·
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.3390/metabo14110622
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.3390/metabo14110622
Background/Objectives: Targeted metabolomics is often criticized for the limited metabolite coverage that it offers. Indeed, most targeted assays developed or used by researchers measure fewer than 200 metabolites. In an effort to both expand the coverage and improve the accuracy of metabolite quantification in targeted metabolomics, we decided to develop a comprehensive liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay that could quantitatively measure more than 700 metabolites in serum or plasma. Methods: The developed assay makes use of chemical derivatization followed by reverse phase LC–MS/MS and/or direct flow injection MS (DFI–MS) in both positive and negative ionization modes to separate metabolites. Multiple reaction monitoring (MRM), in combination with isotopic standards and multi-point calibration curves, is used to detect and absolutely quantify the targeted metabolites. The assay has been adapted to a 96-well plate format to enable automated, high-throughput sample analysis. Results: The assay (called MEGA) is able to detect and quantify 721 metabolites in serum/plasma, covering 20 metabolite classes and many commonly used clinical biomarkers. The limits of detection were determined to range from 1.4 nM to 10 mM, recovery rates were from 80% to 120%, and quantitative precision was within 20%. LC–MS/MS metabolite concentrations of the NIST® SRM®1950 plasma standard were found to be within 15% of NMR quantified levels. The MEGA assay was further validated in a large dietary intervention study. Conclusions: The MEGA assay should make comprehensive quantitative metabolomics much more affordable, accessible, automatable, and applicable to large-scale clinical studies.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.3390/metabo14110622
- OA Status
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- Cited By
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- References
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- OpenAlex ID
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https://doi.org/10.3390/metabo14110622Digital Object Identifier
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A Comprehensive LC–MS Metabolomics Assay for Quantitative Analysis of Serum and PlasmaWork title
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articleOpenAlex work type
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enPrimary language
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2024Year of publication
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2024-11-14Full publication date if available
- Authors
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Lun Zhang, Jiamin Zheng, Mathew Johnson, Rupasri Mandal, M. D. Soler Cruz, Miriam Martínez‐Huélamo, Cristina Andrés‐Lacueva, David S. WishartList of authors in order
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goldOpen access status per OpenAlex
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Metabolomics, Quantitative analysis (chemistry), Chromatography, Computational biology, Chemistry, BiologyTop concepts (fields/topics) attached by OpenAlex
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12Total citation count in OpenAlex
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