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Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Abstract : 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.
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https://hal-univ-bourgogne.archives-ouvertes.fr/hal-01687093
Contributeur : Lnc - Université de Bourgogne <>
Soumis le : jeudi 18 janvier 2018 - 10:42:04
Dernière modification le : mardi 27 octobre 2020 - 14:34:45

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Jean-Paul Pais de Barros, Thomas Gautier, Wahib Sali, Christophe Adrie, Hélène Choubley, et al.. Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay. Journal of Lipid Research, American Society for Biochemistry and Molecular Biology, 2015, 56 (7), pp.1363 - 1369. ⟨10.1194/jlr.D059725⟩. ⟨hal-01687093⟩

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