Targeted eicosanoids lipidomics of exhaled breath condensate in healthy subjects

BackgroundExhaled breath condensate collection is a non-invasive method of sampling the respiratory tract that can be repeated several times in a wide range of clinical settings. Quantitation of non-volatile compounds in the condensate requires highly sensitive analytical methods, e.g. mass spectrometry.ObjectiveTo validate cross-platform measurements of eicosanoids using high performance liquid chromatography or gas chromatography coupled with mass spectrometry in exhaled breath condensate sampled from 58 healthy individuals.MethodsTwenty different eicosanoid compounds, representing major arachidonic acid lipoxygenation and cyclooxygenation pathways were measured using a stable isotope dilution method. We applied a free palmitic acid concentration as a surrogate marker for the condensate dilution factor.ResultsEicosanoids concentrations in the condensates were consistent with their content in other biological fluids. Prostaglandin E2 was the most abundant mediator, represented by its stable metabolite tetranor-PGEM. Prostaglandin D2 products were at low concentration, while hydroxyacids derived from lipoxygenation were abundant. 5-HETE was elevated in current tobacco smokers. Leukotriene B4 has the highest concentration of all 5-LO products. 15-LO analogues of cysteinyl leukotrienes–eoxins were detectable and metabolized to eoxin E4. Two main vascular prostanoids: prostacyclin and thromboxane B2 were present as metabolites. A marker for non-enzymatic lipid peroxidation, 8-iso-PGF2α isoprostane was increased in smokers.ConclusionPresented targeted lipidomics analysis of exhaled breath condensate in healthy subjects justifies its application to investigation of inflammatory lung diseases. Measurements of non-volatile mediators of inflammation in the condensates might characterize disease-specific pathological mechanisms and responses to treatment.