Purpose: To develop a sensitive, reliable and cost-effective bioanalytical method for the pharmacokinetic analysis of rasagiline in human plasma. Method: Rasagiline was extracted by liquid-liquid extraction method and analyzed by reversed-phase high performance liquid chromatography (HPLC) using a mixture of ammonium acetate (pH 5.8) and acetonitrile (55:45, v/v) as mobile phase at a flow rate of 1 mL/min. The separation was performed on a Lichrosphere reverse-phase (RP) C18 column (250 x 4.6

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... , 5 μm particle size) at ambient temperature and rasagiline was detected at a wavelength of 265 nm by ultra-violet UV detection. The method was validated according to European Medicine Agency (EMA) guidelines. Results: The developed method was linear over a concentration range of 0.5 -20 µg/ml with r2 ≥ 0.999 in human plasma. Run time was 10 min with rasagiline peak appearing at 7 min with no interference. Relative recovery and relative standard deviation (RSD) for accuracy and precision were within the acceptable limits prescribed in EMA guidelines. Rasagiline remained stable in human plasma for 24 h at room temperature, after three freeze and thaw cycles and also for 3 months at -20 °C. Conclusion: A simple and reliable method has been successfully developed and validated for the determination of rasagiline concentration in human plasma. This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest