Selective and sensitive method for the determination of metoprolol in human plasma using liquid chromatography coupled with tandem mass spectrometry.

A high-performance liquid chromatography-tandem mass spectrometric method was developed and validated for the determination of metoprolol in human plasma. The analyte and internal standard, nevirapine, were extracted from plasma matrix by liquid-liquid extraction with ethyl acetate. Chromatographic separation was achieved on a C-18 analytical column with an isocratic mobile phase of 15:85 (V/V) 10 mmol L-1 ammonium acetate (pH 5.0)/acetonitrile. The atmospheric pressure chemical ionization technique was used for sample ionization in positive ion mode and enhanced selectivity was achieved by tandem mass spectrometric analysis via two multiple reaction monitoring (MRM) transitions, 268.2 → 116.2 for metoprolol and 267.1 → 226.2 for nevirapine, respectively. The assay was validated for human plasma over a concentration range of 1-200 ng mL-1 with the precision and accuracy ranging from 0.9 to 8.8% and 89.9 to 105.8%, respectively.