Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay.
Fiche publication
Date publication
juillet 2015
Journal
Journal of lipid research
Auteurs
Membres identifiés du Cancéropôle Est :
Dr LAGROST Laurent, Dr PAIS DE BARROS Jean-Paul
Tous les auteurs :
Pais de Barros JP, Gautier T, Sali W, Adrie C, Choubley H, Charron E, Lalande C, Le Guern N, Deckert V, Monchi M, Quenot JP, Lagrost L
Lien Pubmed
Résumé
Quantitation of plasma lipopolysaccharides (LPSs) might be used to document Gram-negative bacterial infection. In the present work, LPS-derived 3-hydroxymyristate was extracted from plasma samples with an organic solvent, separated by reversed phase HPLC, and quantitated by MS/MS. This mass assay was combined with the limulus amebocyte lysate (LAL) bioassay to monitor neutralization of LPS activity in biological samples. The described HPLC/MS/MS method is a reliable, practical, accurate, and sensitive tool to quantitate LPS. The combination of the LAL and HPLC/MS/MS analyses provided new evidence for the intrinsic capacity of plasma lipoproteins and phospholipid transfer protein to neutralize the activity of LPS. In a subset of patients with systemic inflammatory response syndrome, with documented infection but with a negative plasma LAL test, significant amounts of LPS were measured by the HPLC/MS/MS method. Patients with the highest plasma LPS concentration were more severely ill. HPLC/MS/MS is a relevant method to quantitate endotoxin in a sample, to assess the efficacy of LPS neutralization, and to evaluate the proinflammatory potential of LPS in vivo.
Mots clés
Animals, Blood Chemical Analysis, methods, Chromatography, High Pressure Liquid, Female, Horseshoe Crabs, Humans, Lipopolysaccharides, blood, Male, Membrane Proteins, metabolism, Mice, Middle Aged, Tandem Mass Spectrometry
Référence
J. Lipid Res.. 2015 Jul;56(7):1363-9