title: Salting-out assisted extraction method coupled with hydrophilic interaction liquid chromatography for determination of selected β-blockers and their metabolites in human urine.
abstract: In this study, a new analytical method was developed and validated for the simultaneous analysis of β-blockers (metoprolol, propranolol, carvedilol) and their metabolites (5'-hydroxycarvedilol, O-desmethylcarvedilol, α-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 all analytes. 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 β-blockers and metabolites on the five different HILIC columns were examined. The method was linear for concentrations ranging from 0.1 to 8.0μg/mL, with determination coefficients higher than 0.993 for all analytes. The limits of quantification were in the range from 0.1 to 0.2μg/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 β-blockers and their metabolites in human urine samples.
authors.affiliation: Department of Inorganic, Analytical Chemistry and Electrochemistry, Silesian University of Technology, 7M. Strzody Str., 44-100 Gliwice, Poland. Electronic address: sylwia.magiera@polsl.pl.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Magiera
authors.affiliation: Department of Inorganic, Analytical Chemistry and Electrochemistry, Silesian University of Technology, 7M. Strzody Str., 44-100 Gliwice, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Kolanowska
authors.affiliation: Department of Clinical Physiology, University Hospital, SE-581 85 Linköping, Sweden.
authors.forename: Jacek
authors.initials: J
authors.lastname: Baranowski
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D000319:Adrenergic beta-Antagonists / Q000737:chemistry / Q000378:metabolism / Q000652:urine*; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D016014:Linear Models; D059625:Liquid-Liquid Extraction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012965:Sodium Chloride / Q000737:chemistry; D013696:Temperature
chemical_list: D000097:Acetonitriles; D000319:Adrenergic beta-Antagonists; D012965:Sodium Chloride; C032159:acetonitrile
keywords: HILIC; Metabolites; SALLE; β-Blockers