Determination of cysteinyl leukotrienes in exhaled breath condensate: Method combining immunoseparation with LC–ESI-MS/MS

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 879; no. 23; pp. 2220 - 2228
• Main Authors: Syslová, Kamila, Kačer, Petr, Vilhanová, Beáta, Kuzma, Marek, Lipovová, Petra, Fenclová, Zdenka, Lebedová, Jindřiška, Pelclová, Daniela
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.08.2011; Elsevier
• Subjects: Adolescent; Analysis; Analytical, structural and metabolic biochemistry; asthma; Asthma - diagnosis; Biological and medical sciences; Breath Tests - methods; Chromatography, Liquid - methods; Cysteine - analysis; Cysteinyl leukotriene; detection limit; electrospray ionization mass spectrometry; Exhalation; Exhaled breath condensate; Female; freeze drying; Fundamental and applied biological sciences. Psychology; General pharmacology; Humans; Immunoassay - methods; Immunoseparation; LC–MS; leukotrienes; Leukotrienes - analysis; liquid chromatography; Lyophilization; Male; Medical sciences; monitoring; patients; Pharmacology. Drug treatments; pretreatment; Solid phase extraction; Spectrometry, Mass, Electrospray Ionization - methods; Young Adult; Solid phase extraction; Lyophilization; Immunoseparation; Cysteinyl leukotriene; Exhaled breath condensate; LC–MS; Expired air; Freeze drying; Arachidonic acid derivatives; Polyunsaturated fatty acid; HPLC chromatography; Lipids; Determination; Electrospray; Chemical enrichment; Leukotriene; LC-MS; Sample preparation; Eicosanoid; Unsaturated fatty acid; Mass spectrometry; Quantitative analysis
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2011.06.004

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.