Determination of metoprolol and its two metabolites in human plasma and urine by high performance liquid chromatography with fluorescence detection and its application in pharmacokinetics

J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Oct 15:937:60-6. doi: 10.1016/j.jchromb.2013.08.017. Epub 2013 Aug 24.

Abstract

A simple, specific and sensitive HPLC method has been developed for the determination of metoprolol and its two metabolites in human plasma and urine. Separation of metoprolol, α-hydroxymetoprolol, O-desmethylmetoprolol and esmolol (internal standard) was achieved on an Agilent XDB-C18 column (150mm×4.6mm, 5μm) using fluorescence detection with Ex 216nm and Em 312nm. The mobile phase consisted of ACN-H2O-0.1%TFA. The analysis was performed in less than 16min with a flow rate of 0.8mL/min. The assay was linear over the concentration range of 5-600ng/mL and 2.5-300ng/mL for metoprolol and its metabolites, respectively. The LOQ were 5.0 and 2.5ng/mL for plasma and urine, respectively. Good precision and accuracy for metoprolol and its two metabolites were obtained. The extraction recoveries were found to be more than 86.91% both in plasma and urine. At the same time, the method was successfully applied to nine healthy volunteers who had been given an oral tablet of 100mg metoprolol.

Keywords: HPLC-FLD detection; Human plasma and urine; Metoprolol; O-desmethylmetoprolol; Pharmacokinetics; α-Hydroxymetoprolol.

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Linear Models
  • Metoprolol / analogs & derivatives
  • Metoprolol / blood*
  • Metoprolol / pharmacokinetics
  • Metoprolol / urine*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence

Substances

  • Metoprolol