title: 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.
authors.affiliation: 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.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Pais de Barros
authors.affiliation: 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.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gautier
authors.affiliation: 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.
authors.forename: Wahib
authors.initials: W
authors.lastname: Sali
authors.affiliation: Physiology Department, Cochin Hospital, Paris University, Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Adrie
authors.affiliation: LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Choubley
authors.affiliation: LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Charron
authors.affiliation: LipSTIC LabEx, Fondation de Coopération Scientifique Bourgogne-Franche Comté, F-21000 Dijon, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Lalande
authors.affiliation: 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.
authors.forename: Naig
authors.initials: N
authors.lastname: Le Guern
authors.affiliation: 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.
authors.forename: Valérie
authors.initials: V
authors.lastname: Deckert
authors.affiliation: Intensive Care Unit, Melun General Hospital, Melun, France.
authors.forename: Mehran
authors.initials: M
authors.lastname: Monchi
authors.affiliation: 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.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Quenot
authors.affiliation: 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.
authors.forename: Laurent
authors.initials: L
authors.lastname: Lagrost
mesh_terms: D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006737:Horseshoe Crabs; D006801:Humans; D008070:Lipopolysaccharides / Q000097:blood* / Q000737:chemistry / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008875:Middle Aged; D053719:Tandem Mass Spectrometry
chemical_list: D008070:Lipopolysaccharides; D008565:Membrane Proteins; C025357:endotoxin binding proteins
keywords: diagnostic tool; human; inflammation; lipid transfer protein; lipoprotein; liquid chromatography tandem mass spectrometry; mass spectrometry; mouse; sepsis; systemic inflammatory response syndrome