Isocratic High Performance Liquid Chromatographic Method for Determination of Metoprolol and Its Metabolite in Human Urine
Abstract: An isocratic
reversed phase high performance liquid chromatographic (HPLC) method has been
developed for the determination of metoprolol (MET) and α-hydroxymetoprolol
(α-OH MET) in human urine. Analytes were extracted using dichloromethane. The
optimized separations were performed on a Purospher® STAR RP-18e LiChroCART®
(250 x 4.6 mm, 5μm) HPLC column within 12 min by an isocratic elution with 25
mmol/L aqueous potassium dihydrogen phosphate buffer (pH 3.0) containing 15%
(v/v) acetonitrile and 15% (v/v) methanol, and UV detection at 234 nm. The method
was validated for linearity, accuracy, precision and specificity. The
calibration curves for both analytes were linear over the range investigated
(1.0-64.0 μg/mL) with correlation coefficients of 0.999. Relative standard
deviation (RSD) values were below 2%. For all of the analytes, recoveries value
from 98.36±1.68 to 101.32±1.36 % and the limits of detection for MET and α-OH
MET were 0.22 and 0.14 μg/mL, respectively. The method developed has been
demonstrated to be sensitive and reliable for the measurement of MET and α-OH
MET simultaneously in human urine.
Keywords: high performance
liquid chromatography, metoprolol, metabolite, human urine
Penulis: PRI ISWATI UTAMI
Kode Jurnal: jpfarmasidd160439
