title: Β-hydroxymyristic acid as a chemical marker to detect endotoxins in dialysis water.
abstract: An analytical chemical method has been developed for determination of β-hydroxymyristic acid (β-HMA), a component of lipopolysaccharides (LPSs/endotoxins) in dialysis water. In our investigation, the β-HMA component was used as a chemical marker for endotoxin presence in dialysis water because it is available in the molecular subunit (lipid A) and responsible for toxicity. It is the most abundant saturated fatty acid in that subunit. The developed method is based on fluorescence derivatization with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ). A high-performance liquid chromatographic separation of the β-HMA derivative was achieved using an octadecyl silica column in gradient elution. A wide dynamic range of β-HMA was tested and a calibration curve was constructed with accuracy of 90% and variability of less than 10%. The limits of detection and quantification obtained were 2 and 5μM, respectively. The developed method was applied to detect endotoxins in dialysis water by alkaline hydrolysis of LPS using NaOH (0.25M) at 60°C for 2h. After hydrolysis, free acid was detected as its NBD-PZ derivative using high-performance liquid chromatography/mass spectrometry (HPLC/MS). Good recovery rates ranging from 98 to 105% were obtained for β-HMA in dialysis water.
authors.affiliation: Aix Marseille Université, Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie de l'Environnement (FRE 3416), 13331 Marseille, France.
authors.forename: Rupesh K
authors.initials: RK
authors.lastname: Mishra
authors.affiliation: Aix Marseille Université, Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie de l'Environnement (FRE 3416), 13331 Marseille, France.
authors.forename: Fabien
authors.initials: F
authors.lastname: Robert-Peillard
authors.affiliation: Aix Marseille Université, Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie de l'Environnement (FRE 3416), 13331 Marseille, France.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Ravier
authors.affiliation: Aix Marseille Université, Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie de l'Environnement (FRE 3416), 13331 Marseille, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Coulomb
authors.affiliation: Aix Marseille Université, Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie de l'Environnement (FRE 3416), 13331 Marseille, France. Electronic address: jean-luc.boudenne@univ-amu.fr.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Boudenne
mesh_terms: D002138:Calibration; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D006868:Hydrolysis; D008070:Lipopolysaccharides / Q000032:analysis* / Q000737:chemistry; D009227:Myristic Acids / Q000032:analysis* / Q000737:chemistry; D006435:Renal Dialysis; D014867:Water / Q000737:chemistry*
chemical_list: D008070:Lipopolysaccharides; D009227:Myristic Acids; D014867:Water; C009893:beta-hydroxymyristic acid
keywords: Endotoxins; HPLC–LC/MS; Labeling; Lipopolysaccharide; β-Hydroxymyristic acid