dc.contributor.author	Khamis, Mona
dc.contributor.author	Adamko, Darryl
dc.contributor.author	El-Aneed, Anas
dc.date.accessioned	2019-04-30T15:57:11Z
dc.date.available	2019-04-30T15:57:11Z
dc.date.issued	2017-10-09
dc.description.abstract	Obstructive airways inflammatory diseases sometimes show overlapping symptoms that hinder their early and correct diagnosis. Current clinical tests are tedious and are of inadequate specificity in special population such as the elderly and children. Therefore, we are developing tandem mass spectrometric (MS/MS) methods for targeted analysis of urine biomarkers. Recently, proton-nuclear magnetic resonance (1H-NMR) analysis proposed 50 urinary metabolites as potential diagnostic biomarkers among asthma and chronic obstructive pulmonary disease (COPD) patients. Metabolites are divided into 3 groups based on chemical nature. For group 1 (amines and phenols, 19 urinary metabolites), we developed and validated a high performance liquid chromatographic (HPLC)-MS/MS method using differential isotope labeling with dansyl chloride. Method development included the optimization of the derivatization reaction, the MS/MS conditions, and the chromatographic separation. Linearity varied from 2 to 4800 ng/mL and the use of 13C2-labeled derivatives allowed for the correction of matrix effects as well as the unambiguous confirmation of the identity of each metabolite in the presence of interfering isomers in urine. Despite the challenges associated with method validation, the method was fully validated as per the food and drug administration (FDA) and the European medicines agency (EMA) recommendations. Validation criteria included linearity, precision, accuracy, dilution integrity, selectivity, carryover, and stability. Challenges in selectivity experiments included the isotopic contributions of the analyte towards its internal standard (IS), that was addressed via the optimization of the IS concentration. In addition, incurred sample analysis was performed to ensure that results from patient samples are accurate and reliable. The method was robust and reproducible and is currently being applied in a cohort of asthma and COPD patient urine samples for biomarker discovery purposes.	en_US
dc.description.sponsorship	Saskatchewan Health Research Foundation; AllerGen NCE Inc; Canada Foundation for Innovation; College of Pharmacy and Nutrition, University of Saskatchewan	en_US
dc.description.version	Peer Reviewed	en_US
dc.identifier.citation	Mona M. Khamis, Darryl J. Adamko and Anas El-Aneed. (2017), Development of a validated LC- MS/MS method for the quantification of 19 endogenous asthma/COPD potential urinary biomarkers. Analytica Chimica Acta, 989: 45-58.	en_US
dc.identifier.doi	10.1016/j.aca.2017.08.007
dc.identifier.uri	http://hdl.handle.net/10388/11995
dc.language.iso	en	en_US
dc.publisher	Elsevier	en_US
dc.rights	Attribution-NonCommercial-ShareAlike 2.5 Canada	*
dc.rights.uri	http://creativecommons.org/licenses/by-nc-sa/2.5/ca/	*
dc.title	Development of a validated LC- MS/MS method for the quantification of 19 endogenous asthma/COPD potential urinary biomarkers	en_US
dc.type	Postprint	en_US

Development of a validated LC- MS/MS method for the quantification of 19 endogenous asthma/COPD potential urinary biomarkers

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