Development of microflow ultra high performance liquid chromatography-mass spectrometry metabolomic assays for analysis of mammalian biofluids.


Journal

Metabolomics : Official journal of the Metabolomic Society
ISSN: 1573-3890
Titre abrégé: Metabolomics
Pays: United States
ID NLM: 101274889

Informations de publication

Date de publication:
25 Oct 2024
Historique:
received: 24 06 2024
accepted: 10 10 2024
medline: 25 10 2024
pubmed: 25 10 2024
entrez: 25 10 2024
Statut: epublish

Résumé

The application of untargeted metabolomics assays using ultra high performance liquid chromatography-mass spectrometry (UHPLC-MS) to study metabolism in biological systems including humans is rapidly increasing. In some of these studies there is a requirement to collect and analyse low sample volumes of biofluids (e.g. tear fluid) or low cell and tissue mass samples (e.g. tissue needle biopsies). The application of microflow, capillary or nano liquid chromatography (≤ 1.0 mm column internal diameter (i.d.)) theoretically should accomplish a higher assay sensitivity compared to analytical liquid chromatography (2.1-5.0 mm column internal diameter). To date, there has been limited research into microflow UHPLC-MS assays that can be applied to study samples of low volume or mass. This paper presents three complementary UHPLC-MS assays (aqueous C All three assays showed an increase in peak areas and peak widths when applying the 1.0 mm i.d. assays. HILIC assays provide an advantage at lower sample dilutions whereas for reversed phase (RP) assays there was no benefit added. This can be seen in the validation study where a much higher number of compounds were detected in the HILIC assay. RP assays were still appropriate for small volume samples with hundreds of compounds being detected. In summary, the 1.0 mm i.d. column assays are applicable for small volume samples where dilution is required during sample preparation.

Identifiants

pubmed: 39453548
doi: 10.1007/s11306-024-02187-y
pii: 10.1007/s11306-024-02187-y
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

120

Informations de copyright

© 2024. The Author(s).

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Auteurs

Annie J Harwood-Stamper (AJ)

School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.

Caroline A Rowland (CA)

Defence Science and Technology Laboratory, Porton Down, Salisbury, SP4 0JQ, UK.

Warwick B Dunn (WB)

School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK. warwick.dunn@liverpool.ac.uk.
Centre for Metabolomics Research, Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool, L69 7ZB, UK. warwick.dunn@liverpool.ac.uk.

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