title: Enantioselective determination of metoprolol and its metabolites in human urine high-performance liquid chromatography with fluorescence detection (HPLC-FLD) and tandem mass spectrometry (MS/MS).
abstract: A sensitive, stereoselective assay using solid phase extraction and high-performance liquid chromatography (HPLC) with fluorescence detection (FLD) was developed and validated for the analysis of enantiomers of metoprolol and its metabolites (α-hydroxymetoprolol, O-desmethylmetoprolol). Chiral separation was achieved using a CHIRALCEL OD-RH column, packed with cellulose tris-(3,5-dimethylphenyl-carbamate) stationary phase, employing a mobile phase composed by a mixture of 0.2% diethylamine in water and acetonitrile in gradient elution mode. Linear calibration curves were obtained over the range of 0.025-2.0μg/mL (R(2)>0.994) in urine for both enantiomers of metoprolol and its metabolites with quantitation limit of 0.025μg/mL. Intra and inter-day precision and accuracy were below 15% for both metoprolol and metabolites enantiomers. The recovery of enantiomer of metoprolol and its metabolite was greater than 68.0%, utilizing a SPE procedure. The method was tested with urine quality control samples and human urine fractions after administration of 50mg rac-metoprolol.
authors.affiliation: Department of Inorganic, Analytical Chemistry and Electrochemistry, Silesian University of Technology, 7 M. Strzody Str., 44-100 Gliwice, Poland. Electronic address: irena.baranowska@polsl.pl.
authors.forename: Irena
authors.initials: I
authors.lastname: Baranowska
authors.affiliation: Department of Inorganic, Analytical Chemistry and Electrochemistry, Silesian University of Technology, 7 M. Strzody Str., 44-100 Gliwice, Poland.
authors.forename: Weronika
authors.initials: W
authors.lastname: Adolf
authors.affiliation: Department of Inorganic, Analytical Chemistry and Electrochemistry, Silesian University of Technology, 7 M. Strzody Str., 44-100 Gliwice, Poland.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Magiera
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000652:urine*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D008790:Metoprolol / Q000652:urine*; D012015:Reference Standards; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000319:Adrenergic beta-Antagonists; D008790:Metoprolol
keywords: Chiral separation; Enantiomers; Metoprolol; Urine samples