Identification and relative quantification of fatty acids in Escherichia coli membranes by gas chromatography/mass spectrometry

Abstract

The lipids that are essential to the functioning of the bacterial membrane exist in hundreds of different forms. The reasons for this diversity are far from clear but are presumably related to the roles of these lipids in both facilitating enzymic activities and generating proteolipid domains. A full understanding of bacterial physiology therefore requires characterization of lipids in different strains in a variety of environmental conditions. This characterization then becomes the basis for lipidomics, the lipid aspect of the growing field of metabolomics. To exploit the power of derivatization chemistry and of gas chromatography/mass spectrometry (GC/MS) and tandem mass spectrometry (MS/MS) for metabolomics studies, we report here the development of various GC/MS electron ionization (EI) and negative and positive chemical ionization (CI) methods for the identification and, for the first time, the relative quantification of fatty acids present in extracts from membranes of a laboratory strain of Escherichia coli. They consist of seven saturated fatty acids (C10:0, C12:0, C14:0, C15:0, C16:0, C17:0 and C18:0) and six unsaturated fatty acids (C16:1, cyC17:0 plus two isomers of C18:1, C18:2 and cyC19:0).