Salting-out assisted extraction method coupled with hydrophilic interaction liquid chromatography for determination of selected beta-blockers and their metabolites in human urine
2016 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 1022, 93-101 p.Article in journal (Refereed) PublishedText
In this study, a new analytical method was developed and validated for the simultaneous analysis of beta-blockers (metoprolol, propranolol, carvedilol) and their metabolites (5-hydroxycarvedilol, O-desmethylcarvedilol, alpha-hydroxymetoprolol, O-desmethylmetoprolol, 5-hydroxypropranolol) in human urine. A salting-out assisted liquid-liquid extraction (SALLE) procedure was used for sample preparation. Several parameters affecting the extraction efficiency and method sensitivity including the type and volume of the extraction solvent, the type and quantity of the inorganic salt, extraction time and sample pH were investigated. Hydrophilic interaction liquid chromatography-ultraviolet detection (HILIC-UV) was used for the determination of allanalytes. During method development, the effects of mobile phase components (type, pH, concentration of salt, organic modifier type and content, flow rate, column temperature) on the retention and separation of beta-blockers and metabolites on the five different HILIC columns were examined. The method was linear for concentrations ranging from 0.1 to 8.0 mu g/mL, with determination coefficients higher than 0.993 for all analytes. The limits of quantification were in the range from 0.1 to 0.2 mu g/mL. Intra- and inter-day precision ranged from 0.1 to 8.9%, and accuracy was within +/- 13% interval for all analytes. Under the optimized conditions, extraction efficiency was greater than 83.4% for determined compounds. The validated method was then applied to the measurement of beta-blockers and their metabolites in human urine samples. (C) 2016 Elsevier B.V. All rights reserved.
Place, publisher, year, edition, pages
ELSEVIER SCIENCE BV , 2016. Vol. 1022, 93-101 p.
HILIC; SALLE; beta-Blockers; Metabolites
IdentifiersURN: urn:nbn:se:liu:diva-129486DOI: 10.1016/j.jchromb.2016.04.010ISI: 000376545800014PubMedID: 27085018OAI: oai:DiVA.org:liu-129486DiVA: diva2:940529