Development of rapid and high-throughput LC-MS/MS method for quantification of olprinone in rabbit plasma

A simple, highly sensitive and rapid method for quantification of olprinone (phosphodiesterase 3 inhibitor) in rabbit plasma using liquid chromatography-tandem mass spectrometry with electrospray was developed. An aliquot of 50 mu L of plasma sample was cleaned up and extracted using Ostro (TM) 96-well plate followed by dilution. Chromatographic separation of olprinone and olprinone-d3 was carried out on a CORTECS (R) T3 column within 3 min. Detection was achieved using a triple quadrupole mass spectrometer employing electrospray ionization operated in positive ion multiple reaction monitoring mode using the transitions m/z 251.07 -> m/z 155.06 and m/z 254.21 -> m/z 158.10 for olprinone and olprinone-d3, respectively. The method was validated according to US Food and Drug Administration guideline for bioanalytical methods, and showed excellent linearity in the range 10.0-2000.0 ng/mL with coefficient of determination >0.99. The intra- and inter-day precisions (CV) were <5.1% and the accuracies were within the range 99.7-103.2% at all quality control concentrations. Furthermore, olprinone was stable under various stability conditions. The developed method was used for quantification of olprinone in rabbit plasma after its intravenous administration at the dose of 1 mg/kg in order to better understand the metabolism of olprinone in a rabbit model of lung injury.