title: Quantification of bacterial lipopolysaccharides (endotoxin) by GC-MS determination of 3-hydroxy fatty acids.
abstract: A GC-MS method for the quantification of bacterial lipopolysaccharides (LPS, endotoxin) is presented. After hydrolytic cleavage of 3-hydroxy fatty acids (3-OH FAs) from the lipid A region of LPS, derivatisation of both the hydroxyl and the carboxyl group was performed in one step with a mixture of methyl-bis(trifluoracetamide)(MBTFA) and N-methyl-N-(tert-butyldimethylsilyl)trifluoracetamide (MTBSTFA). Using GC-MS in the EI mode with selected ion monitoring (SIM) for analysis, baseline separation of 3-OH FAs (and of possibly interfering 2-OH FAs) was achieved. The sensitivity of the method (LOD 7-50 pg/injection for the different 3-OH FAs investigated) allows for the efficient quantification of LPS in occupational and environmental samples. Degradation of 3-OH FAs as well as of their derivatives during sample preparation and GC-MS separation as a possible source of errors in analytical methods based on 3-OH FA determination is reported for the first time. Thermal elimination of water from the underivatised 3-OH FAs and of trifluoroacetic acid from the derivatives was identified as the cause of degradation. The resulting alpha,beta-unsaturated compounds showing the same mass spectra as the 3-OH FA derivatives were detected as more or less prominent satellite peaks. By using alkaline instead of acidic hydrolysis and cool on-column instead of split/splitless injection, elimination was reduced to an acceptable level.
authors.affiliation: Institute of Occupational Medicine, University of Münster, Münster D-48149, Germany.
authors.forename: Norbert
authors.initials: N
authors.lastname: Binding
authors.forename: Sabine
authors.initials: S
authors.lastname: Jaschinski
authors.forename: Sabine
authors.initials: S
authors.lastname: Werlich
authors.forename: Stefan
authors.initials: S
authors.lastname: Bletz
authors.forename: Ute
authors.initials: U
authors.lastname: Witting
mesh_terms: D004784:Environmental Monitoring / Q000379:methods; D005227:Fatty Acids / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry; D008070:Lipopolysaccharides / Q000032:analysis*; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D005227:Fatty Acids; D008070:Lipopolysaccharides