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© Laurent Debarbieux
Bactériophages sur la bactérie Escherichia coli 0104:H4 souche 55989.
Publication : Journal of chromatography. B, Analytical technologies in the biomedical and life sciences

Improving the intestinal lipidome coverage in a gnotobiotic mouse model using UHPLC-MS-based approach through optimization of mobile phase modifiers and column selection.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Journal of chromatography. B, Analytical technologies in the biomedical and life sciences - 15 Jul 2024

Selmi H, Walker A, Debarbieux L, Schmitt-Kopplin P

Link to Pubmed [PMID] – 38901159

Link to DOI – 10.1016/j.jchromb.2024.124188

J Chromatogr B Analyt Technol Biomed Life Sci 2024 Jul; 1242(): 124188

Lipidomics is focusing on the screening of lipid species in complex mixtures using mass spectrometry-based approaches. In this work, we aim to enhance the intestinal lipidome coverage within the Oligo-Mouse-Microbiota (OMM12) colonized mouse model by testing eight mobile phase conditions on five reversed-phase columns. Our selected mobile phase modifiers included two ammonium salts, two concentrations, and the addition of respective acids at 0.1 %. We compared two columns with hybrid surface technology, two with ethylene bridged hybrid technology and one with core-shell particles. Best performance was attained for standards and intestinal lipidome, using either ammonium formate or acetate in ESI(+) or ammonium acetate in ESI(-) for all column technologies. Notably, a concentration of 5 mM ammonium salt showed optimal results for both modes, while the addition of acids had a negligible effect on lipid ionization efficiency. The HST BEH C18 column improved peak width and tailing factor parameters compared to other technologies. We achieved the highest lipid count in colon and ileum content, including ceramides, phosphatidylethanolamines and phosphatidylcholines, when using 5 mM ammonium acetate in ESI(-). Conversely, in ESI(+) 5 mM ammonium formate demonstrated superior coverage for diacylglycerols and triacylglycerols.