Journal
CARBOHYDRATE POLYMERS
Volume 87, Issue 4, Pages 2730-2734Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carbpol.2011.11.073
Keywords
LPS determination; GC-MS; Polysaccharides; beta-Glucan; Nitric oxide pathway
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Funding
- Brazilian agency FINEP (PRONEX-CARBOIDRATOS, PADCT II/SBIO)
- Brazilian agency CAPES-REUNI (Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior)
- Brazilian agency Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)
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The most common method used to quantify lipopolysaccharide (LPS) in polysaccharide samples is the Limulus amebocyte lysate (LAL) test. It is a very sensitive and simple, although not accurate with samples containing carbohydrates, such as widely distributed (1 -> 3)-linked beta-glucans. Another method, the Polymyxin B assay, suffers interference with samples containing negatively charged polysaccharides. We have now developed a method to detect and quantify LPS in carbohydrate-containing samples, using GC-MS of derived acetylated 3-OH fatty acid methyl esters. The method proved to be robust, highly specific and sensitive, allowing detection of LPS at 1 ng, 100 times less than the amount of LPS frequently used as positive control in immunological experiments. In order to demonstrate the applicability of the method, 14 polysaccharide samples were analyzed. On two of them, the presence of LPS was detected at concentrations of 16.1 and 12.7 ng/300 mu g polysaccharide. (C) 2011 Elsevier Ltd. All rights reserved.
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