LC–MS/MS determination of pramipexole on rat dried blood spots: A pharmacokinetic study

Abstract A simple and selective liquid chromatography–tandem mass spectrometric method for determination of pramipexole on rat dried blood spots was developed and validated. Chromatographic separation was achieved on a Synergy polar-RP column using 10 mM ammonium acetate and methanol (50:50, v/v) as mobile phase in an isocratic mode of elution at a flow rate of 1.0 mL/min. LC–MS was performed in a positive ion electro spray ionization mode and the MS/MS ion transitions 212.10 → 153.03 for PRX and 198.10 → 153.03 for internal standard (2-amino-4,5,6,7-tetrahydro-6-ethyl-amino-benzthiazole) were monitored. The developed method exhibited a linear dynamic range over 100–5000 pg/mL for PRX on dried blood spots. The overall extraction recovery of PRX from DBS was 96.7%. The intra- and inter-day accuracy and precision were within the pre-defined limits of ≤15% at all concentrations. Influence of hematocrit and spot volume on dried blood spot was also evaluated and found to be well within the acceptable limits. The method was successfully applied to pharmacokinetic studies of PRX in rats.