Comparison of three GC/MS methodologies for the analysis of fatty acids in Sinorhizobium meliloti: Development of a micro-scale, one-vial method

Three protocols for fatty acid analysis in Sinorhizobium meliloti were improved by the addition of a number of standards/controls and a silylation step which allowed the determination of recoveries, extents of conversion of lipids to fatty acid methyl esters (FAMEs) and extents of side reactions. Basic hydrolysis followed by acid-catalyzed methylation and transmethylation with sodium methoxide, were the best for the analysis of 3-hydroxy- and cyclopropane fatty acids, respectively. A micro-scale, one-vial method that employed sodium methoxide/methanol was equally efficient and on a 1000-fold smaller scale than standard methods. Because this method avoids aqueous extractions, 3-hydroxybutanoic acid was detected as its trimethylsilyloxy methyl ester along with FAMEs.