Immunomagnetic molecular probe with UHPLC-MS/MS: A promising way for reliable bronchial asthma diagnostics based on quantification of cysteinyl leukotrienes

0425848 - MBÚ 2014 RIV NL eng J - Článek v odborném periodiku
Syslová, K. - Boehmová, A. - Demirbag, E. - Šimková, K. - Kuzma, Marek - Pelclová, D. - Sedlák, V. - Čáp, P. - Martásek, P. - Kačer, P.
Immunomagnetic molecular probe with UHPLC-MS/MS: A promising way for reliable bronchial asthma diagnostics based on quantification of cysteinyl leukotrienes.
Journal of Pharmaceutical and Biomedical Analysis. 81-82, AUG 2013 (2013), s. 108-117. ISSN 0731-7085. E-ISSN 1873-264X
Institucionální podpora: RVO:61388971
Klíčová slova: Cysteinyl leukotrienes * Immunomagnetic separation * UHPLC-MS
Kód oboru RIV: CB - Analytická chemie, separace
Impakt faktor: 2.829, rok: 2013

A sensitive and precise method for simultaneous quantification of cysteinyl leukotrienes (=cys LTs) - leukotriene C-4 (=LTC4), leukotriene D-4 (=LTD4) and leukotriene E-4 (=LTE4) - essential biomarkers of bronchial asthma present in exhaled breath condensate (=EBC) was developed. An immunomagnetic molecular probe was prepared by anchoring cysteinyl leukotrienes antibody on the surface of functionalized monodispersed magnetic particles and used to selectively isolate cys LTs from biological matrices - EBC, plasma and urine. Immobilization and the immunoaffinity capture procedures were optimized to maximize the amount of separated cys LTs, which were detected "off-beads" after acidic elution by UHPLC-ESI-MS/MS operated in a multiple reaction monitoring mode. The developed method was characterized with high precision <= 13.6% (intra-day precision determined as RSD) and <= 14.5% (inter-day precision determined as RSD), acceptable accuracy <= 18.5% (determined as RE), and high recovery of immunoseparation (>= 93.1%) in aforementioned biological matrices. The applicability of the method was demonstrated on EBC, plasma and urine clinical samples of patients with various subtypes of bronchial asthma (occupational, steroid-resistant, moderate with and without corticosteroids therapy) and healthy subjects where reasonable differences in cys LTs concentration levels were found
Trvalý link: http://hdl.handle.net/11104/0231638