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Determination of cysteinyl leukotrienes in exhaled breath condensate: method combining immunoseparation with LC-ESI-MS/MS
Kamila Syslová, Petr Kačer, Beáta Vilhanová, Marek Kuzma, Petra Lipovová, Zdenka Fenclová, Jindřiška Lebedová, Daniela Pelclová
Language English Country Netherlands
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
Grant support
NS10298
MZ0
CEP Register
- MeSH
- Asthma diagnosis MeSH
- Chromatography, Liquid methods MeSH
- Cysteine analysis MeSH
- Breath Tests methods MeSH
- Spectrometry, Mass, Electrospray Ionization methods MeSH
- Immunoassay methods MeSH
- Leukotrienes analysis MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Exhalation MeSH
- Check Tag
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
A rapid and precise method for the identification and quantification of cysteinyl leukotrienes (leukotriene C(4), leukotriene D(4) and leukotriene E(4)), essential markers of bronchial asthma, in exhaled breath condensate was developed. The protocol consists of immunoaffinity separation and a detection step, liquid chromatography combined with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In particular, the selected reaction monitoring mode was used for its extremely high degree of selectivity and the stable-isotope-dilution assay for its high precision of quantification. The developed method was characterized with a high precision (≤ 7.7%, determined as RSD), an acceptable accuracy (90.4-93.7%, determined as recovery), a low limit of detection (≤ 2 pg/ml EBC) and a low limit of quantification (≤ 10 pg/ml EBC). It was compared to other simple, clinically appropriate combinations of pre-treatment methods (solid phase extraction and lyophilization) with LC/MS. Finally, the method (a combination of immunoaffinity separation with LC-MS) was successfully tested in a clinical study where a significant difference was found in the concentration levels of cysteinyl leukotrienes between patients with occupational bronchial asthma and healthy subjects.
Department of Occupational Medicine 1st Medical Faculty Charles University Prague Czech Republic
References provided by Crossref.org
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