UPLC-MS/MS Method for Simultaneous Quantification of Pramipexole, Ropinirole and Rasagiline In Human Plasma and Its Application to A Pharmacokinetic Study

A selective, sensitive and rapid UPLC-MS/MS method has been developed and validated for simultaneous quantification of pramipexole, ropinirole and rasagiline in human plasma using trimetazidine as internal standard (IS). The analytes and IS were extracted from 200 µL of plasma by solid phase extraction technique using strata cartridges which offers high sensitivity, wide linearity without interferences form endogenous matrix components. Chromatographic separation was achieved in 3.00 min run time on a Synergi Polar RP column using a 5 mM ammonium acetate/methanol mobile phase in gradient mode. The quantification of target compounds was performed in a positive electrospray ionization mode and multiple reaction monitoring (MRM). The proposed method was validated over the concentration ranges of 5-50000 pg/mL for each analyte. The intra- and inter-day precision and accuracy results were acceptable as per FDA guidelines. Stability of compounds were established in a battery of stability studies, i.e. bench top, auto sampler, dry extract and long term storage stability as well as freeze-thaw cycles. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic studies.