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Journal of Lipid Research, Vol. 46, 1773-1778, August 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology

Methods

Microextraction of bacterial lipid A: easy and rapid method for mass spectrometric characterization

Asmaa El Hamidi^*, Alina Tirsoaga^*,, Alexey Novikov^*, Ahmed Hussein^* and Martine Caroff^1,*

^* Equipe Endotoxines, Unité Mixte de Recherche 8619 du Centre National de la Recherche Scientifique, Institut de Biochimie, Biophysique Moléculaire et Cellulaire, Université de Paris-Sud, 91405 Orsay, France
Department of Physical Chemistry, University of Bucharest, Bucharest, Romania

Published, JLR Papers in Press, June 1, 2005. DOI 10.1194/jlr.D500014-JLR200

¹ To whom correspondence should be addressed. e-mail: martine.caroff{at}bbmpc.u-psud.fr

Endotoxins (lipopolysaccharides) are the main components of Gram-negative bacterial outer membranes. A quick and simple way to isolate their lipid region (lipid A) directly from whole bacterial cells was devised. This method using hot ammonium-isobutyrate solvent was applied to small quantities of cells and proved to be indispensable when a rapid characterization of lipid A structure by mass spectrometry was required. Biological activities of endotoxins are directly related to the lipid A structures, which vary greatly with cell growth conditions.

This method is suitable for rough- and smooth-type bacteria and very efficient for screening variations in lipid A structures. Data are acquired in a few hours and avoid the use of phenol in extraction.

Abbreviations: Kdo, 2-keto-3-deoxyoctonate; MALDI, matrix-assisted laser desorption/ionization; LPS, lipopolysaccharide; OS, oligosaccharide; PS, polysaccharide

Supplementary key words endotoxin • lipopolysaccharide • hydrolysis

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