Development and validation of a sensitive UHPLC–MS/MS method for quantitation of prucalopride in rat plasma and its application to pharmacokinetics study

A rapid, sensitive, selective and accurate ultra high performance liquid chromatography with tandem mass spectrometry (UHPLC–MS/MS) method was developed and validated for the quantitation of prucalopride in rat plasma using carbamazepine as an internal standard (IS). Separation was achieved on a Waters ACQUITY UHPLC ® HSS C 18 column (2.1   mm   ×   50   mm, 1.8   μm) column with a gradient mobile phase consisting of acetonitrile–water (containing 0.1% formic acid) as mobile phase at a flow rate of 0.2   mL/min. Prucalopride and IS were monitored using positive electrospray triple quadrupole mass spectrometer (Waters Xevo TQD) via multiple reaction monitoring (MRM) mode. The monitored transitions were set at m / z 367.99   →   195.89 and m / z 236.97   →   194.04 for prucalopride and IS, respectively. The achieved lower limit of quantitation was 0.1   ng/mL. The validated method had an excellent linearity in the range of 0.1–100   ng/mL (r   >   0.996). The intra- and inter-day precisions were both   ≤   7.8% for prucalopride and IS, and the average intra- and inter-day accuracies ranged from −3.0% to 8.5%. Extraction recoveries at three levels QC concentrations were in the range of 90.0–110.0% for prucalopride and 99.6% for IS. Matrix effects were found to be acceptable. The validated assay was successfully applied to a pharmacokinetic study of prucalopride following oral administration of 0.25, 0.5, 1.0   mg/kg to female and male rats respectively.