|Development and Validation of HPLC Method for Quantification of Rasagiline in Human Plasma by Using Fluorescent Detector
|Challenges in Quantitative Analysis
|Dr Rabiea Bilal
|University of Health Sciences Lahore
Abstract Information :
Rasagiline mesylate (RM) is currently approved as initial monotherapy or adjunct therapy to levodopa for treatment of idiopathic Parkinson's disease. In order to perform the pharmacokinetic analysis of rasagiline, a sensitive and reliable bioanalytical method is inevitable. This HPLC method is developed and validated for the quantification of rasagiline in human plasma. Rasagiline was extracted by a liquid-liquid extraction method and analyzed on HPLC by using a mixture of ammonium acetate (pH 5.8) and acetonitrile (55:45) as mobile phase at a flow rate of 1 mL/min. The separation was performed on Lichrosphere C18 column (250 x 4.6 mm, 5 µm particle size) at room temperature and rasagiline was detected by using fluorescent detector at 265 nm wavelength. The method was validated by following European Medicine Agency (EMA) guideline. The developed method was linear over the concentration range of 1-35 mcg/L with r2 ≥ 0.999 while the lower limit of quantification was 1 mcg/L. No interfering peak was observed at the retention time of rasagiline which ensures the selectivity. The values of percentage relative recovery and relative standard deviation (RSD%) for accuracy and precision were within the acceptable limit described by EMA guideline. Rasagiline was proved stable in human plasma for 24 h at room temperature, after freeze and thaw cycle and also for more than 3 month at -20°C. The developed method is sensitive and reliable for the quantification of rasagiline in human plasma for its subsequent application in pharmacokinetic analysis.