GC-MS detection and quantification of lipopolysaccharides in polysaccharides through 3-O-acetyl fatty acid methyl esters

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• 作者：Arquimedes Paixã ; o de Santana-Filho ; Guilhermina Rodrigues Noleto ; Philip Albert James Gorin ; Lauro Mera de Souza ; Marcello Iacomini ; Guilherme Lanzi Sassaki ; ^{sassaki@ufpr.br}
• 关键词：LPS ; lipopolysaccharide ; LAL ; limulus amebocyte lysate ; NO ; nitric oxide ; TNF-¦Á ; tumor necrosis factor-alpha ; IL-1 ; interleukin-1 ; 3-OH-FAs ; 3-hydroxy fatty acids ; KDO ; keto-3deoxy-octulosonic acid ; FAME ; fatty acid methyl ester ; SIE ; selected ion extrac
• 刊名：Carbohydrate Polymers
• 出版年：2012
• 出版时间：1 March, 2012
• 年：2012
• 卷：87
• 期：4
• 页码：2730-2734
• 全文大小：261 K

文摘

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 ?#xA0;3)-linked ¦Â-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 ¦Ìg polysaccharide.