Validation of 8-isoprostane and prostaglandin E 2 measurements in exhaled breath condensate

1 November 2003

journal article
clinical trial
Published by Springer Nature in Inflammation Research

Vol. 52 (12) , 502-507
https://doi.org/10.1007/s00011-003-1212-6

Abstract

Objective:To qualitatively validate radioimmunoassays for 8-isoprostane and prostaglandin (PG) E₂ in exhaled breath condensate. 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 (RP-HPLC). The eluted fractions were assayed for 8-isoprostane-like immunoreactivity and PGE₂-like immunoreactivity by radioimmunoassays. In addition, simultaneous measurements of exhaled breath condensate unextracted samples with two anti-8-isoprostane and anti-PGE₂ sera with different cross-reactivity were performed. Results:A single peak of 8-isoprostane-like immunoreactivity and PGE₂-like immunoreactivity co-eluting with 8-isoprostane (retention time: 13 min) and PGE₂ (retention time: 21 min) standards, respectively, was identified by radioimmunoassays. Testing with two different antisera showed similar results for both 8-isoprostane-like immunoreactivity (limits of agreement = 4.5 pg/ml and – 4.1 pg/ml, n = 12) and PGE₂-like immunoreactivity (limits of agreement = 6.1 pg/ ml and – 6.1 pg/ml, n = 12). Conclusion: This study provides evidence for the specificity of the radioimmunoassays for 8-isoprostane and PGE₂ in exhaled breath condensate. This is critical for proposing these markers as a non-invasive way for monitoring airway inflammation.

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