EIA and GC/MS analysis of 8-isoprostane in EBC of children with problematic asthma

S. Carraro, P. E. Cogo, I. Isak, M. Simonato, M. Corradi, V. P. Carnielli, E. Baraldi

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Abstract

Asthmatic airways are characterised by enhanced oxidative stress, which can be studied by measuring biomarkers, such as 8-isoprostane. The aims of the present study were: 1) to measure the concentrations of 8-isoprostane in exhaled breath condensate (EBC) and urine of children with problematic and well-controlled asthma; 2) to compare the concentrations of 8-isoprostane measured by gas chromatographic/negative ion chemical ionisation mass spectrometry (GC/NICI-MS) and by an enzymatic immunoassay (EIA). We recruited 20 asthmatic allergic children, 13 with well-controlled asthma and seven with problematic asthma. They underwent exhaled nitric oxide measurements and spirometry, and both EBC and urine samples were collected. 8-isoprostane was measured in EBC by GC/NICI-MS and EIA. 8-isoprostane concentrations in EBC were significantly higher in children with problematic asthma than in children with well-controlled asthma (p=0.01). An acceptable reproducibility emerged between GC/NICI-MS and EIA (coefficient of reproducibility 11.5 pg·mL -1). 8-isoprostane levels measured in urine did not correlate with those measured in EBC. We showed that 8-isoprostane in EBC was significantly increased in children with problematic asthma, suggesting a role for oxidative stress in this asthma phenotype. In addition we found an acceptable reproducibility of EIA compared to GC/NICI-MS, even if the latter method had higher accuracy. Copyright

Original languageEnglish
Pages (from-to)1364-1369
Number of pages6
JournalEuropean Respiratory Journal
Volume35
Issue number6
DOIs
Publication statusPublished - Jun 2010

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Keywords

  • 8-Isoprostane
  • Children
  • Exhaled breath condensate
  • Mass spectrometry
  • Problematic asthma
  • Urine

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Carraro, S., Cogo, P. E., Isak, I., Simonato, M., Corradi, M., Carnielli, V. P., & Baraldi, E. (2010). EIA and GC/MS analysis of 8-isoprostane in EBC of children with problematic asthma. European Respiratory Journal, 35(6), 1364-1369. https://doi.org/10.1183/09031936.00074909