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

Autor:	Pais de Barros JP; INSERM, LNC UMR866, F-21000 Dijon, France LNC UMR866, University Bourgogne Franche-Comté, F-21000 Dijon, France LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France., Gautier T; INSERM, LNC UMR866, F-21000 Dijon, France LNC UMR866, University Bourgogne Franche-Comté, F-21000 Dijon, France LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France., Sali W; INSERM, LNC UMR866, F-21000 Dijon, France LNC UMR866, University Bourgogne Franche-Comté, F-21000 Dijon, France LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France., Adrie C; Physiology Department, Cochin Hospital, Paris University, Paris, France., Choubley H; LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France., Charron E; LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France., Lalande C; LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France., Le Guern N; INSERM, LNC UMR866, F-21000 Dijon, France LNC UMR866, University Bourgogne Franche-Comté, F-21000 Dijon, France LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France., Deckert V; INSERM, LNC UMR866, F-21000 Dijon, France LNC UMR866, University Bourgogne Franche-Comté, F-21000 Dijon, France LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France., Monchi M; Intensive Care Unit, Melun General Hospital, Melun, France., Quenot JP; INSERM, LNC UMR866, F-21000 Dijon, France LNC UMR866, University Bourgogne Franche-Comté, F-21000 Dijon, France LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France Intensive Care Unit University Hospital of Dijon, F-21000 Dijon, France., Lagrost L; INSERM, LNC UMR866, F-21000 Dijon, France LNC UMR866, University Bourgogne Franche-Comté, F-21000 Dijon, France LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France Clinical Research Department, University Hospital of Dijon, F-21000 Dijon, France.
Jazyk:	angličtina
Zdroj:	Journal of lipid research [J Lipid Res] 2015 Jul; Vol. 56 (7), pp. 1363-9. Date of Electronic Publication: 2015 May 28.
DOI:	10.1194/jlr.D059725
Abstrakt:	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. (Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.)
Databáze:	MEDLINE
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