Validation of leukotriene B₄ measurements in exhaled breath condensate

Abstract:

Objective: To qualitatively validate an enzyme immunoassay to measure leukotriene B₄ in exhaled breath condensate. Exhaled breath condensate is a new non-invasive method to monitor airway inflammation.¶Subjects: Twenty-two subjects with different lung diseases attended the outpatient clinic on one occasion for exhaled breath condensate collection.¶Methods: Samples were pooled together and purified by reverse-phase high-performance liquid chromatography. The fractions eluted were assayed for leukotriene B₄ by enzyme immunoassay.¶Results: A single peak of leukotriene B₄-like immunoreactivity co-eluting with leukotriene B₄ standard (retention time: 24 min) was identified by enzyme immunoassay. Reverse phase-high performance liquid chromatography peak of leukotriene B₄ was clearly separated from those of 6-trans- leukotriene B₄ (retention time: 14 min) and leukotriene B₅ (retention time: 18 min) for which the antiserum used in the enzyme immunoassay had the highest cross-reactivity. Leukotriene B₄ recovery was 64%.¶Conclusions: This study provides evidence for the presence of leukotriene B₄ in the exhaled breath condensate and the specificity of the enzyme immunoassay used.