title: Validation of a high performance liquid chromatography-tandem mass spectrometry method for β-hydroxy fatty acids as environmental markers of lipopolysaccharide.
abstract: The most commonly analytical method used for the assessment of the occupational exposure to airborne endotoxins is the LAL test, but other methods are reported in the literature. The objective of this work was to develop and validate a method using HPLC coupled to triple quadrupole mass spectrometer for the quantitative determination of β-hydroxy-dodecanoic and β-hydroxy-tetradecanoic acids as chemical markers of lipopolysaccharide in Gram-negative strains. Matrix effect was significant but compensated by the use of internal standard; accuracy was always more than 85% and variability always less than 10%. LOD and LLOQ were respectively 2.5 μg/l and 10 μg/l for β-hydroxy-dodecanoic acid, 1 μg/l and 5 μg/l for β-hydroxy-tetradecanoic acid. The analysis of different bacterial strains showed that samples contain β-hydroxy-dodecanoic and β-hydroxy-tetradecanoic acids in variable ratio, and therefore the sum of their concentrations was considered the chemical marker. The method showed a significant correlation of results with those of LAL-test on four bacterial strains.
authors: INAIL Research, Department of Occupational Hygiene, via Fontana Candida, 1-00040 Monteporzio Catone (RM), Italy.
authors: Alessandra
authors: A
authors: Chiominto
authors: INAIL Research, Department of Occupational Hygiene, via Fontana Candida, 1-00040 Monteporzio Catone (RM), Italy.
authors: Anna Maria
authors: AM
authors: Marcelloni
authors: INAIL Research, Department of Occupational Medicine, via Fontana Candida, 1-00040 Monteporzio Catone (RM), Italy. Electronic address: g.tranfo@inail.it.
authors: Giovanna
authors: G
authors: Tranfo
authors: INAIL Research, Department of Occupational Hygiene, via Fontana Candida, 1-00040 Monteporzio Catone (RM), Italy.
authors: Emilia
authors: E
authors: Paba
authors: INAIL Research, Department of Occupational Medicine, via Fontana Candida, 1-00040 Monteporzio Catone (RM), Italy.
authors: Enrico
authors: E
authors: Paci
mesh_terms: D001419:Bacteria / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005227:Fatty Acids / Q000032:analysis*; D005658:Fungi / Q000737:chemistry; D007700:Kinetics; D008070:Lipopolysaccharides / Q000032:analysis* / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D005227:Fatty Acids; D008070:Lipopolysaccharides
keywords: Endotoxin; HPLC-MS/MS; LAL test; Lipopolysaccharide; β-Hydroxy-dodecanoic acid; β-Hydroxy-tetradecanoic acid