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

作者：Syslova Kamila; Kacer Petr^*; Vilhanova Beata; Kuzma Marek; Lipovova Petra; Fenclova Zdenka; Lebedova Jindriska; Pelclova Daniela

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2011, 879(23): 2220-2228.

DOI：10.1016/j.jchromb.2011.06.004

• Cysteinyl leukotriene
• Exhaled breath condensate
• Immunoseparation
• Solid phase extraction
• Lyophilization
• LC-MS

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.

• 出版日期2011-8-1