DEVELOPMENT AND VALIDATION OF HPLC METHOD FOR DETERMINATION OF TRACE LEVEL POTENTIAL GENOTOXIC CHLORO IMPURITY IN RASAGILINE MESYLATE

ABSTRACT

A rapid, sensitive and selective High performance liquid chromatography (HPLC) method was developed and validated for the trace analysis of N-(2-Chloroallyl)-1-(R)-aminoindan, a potential genotoxic impurity in Rasagiline mesylate drug substances. HPLC column used was Sunfire C18 (250 mm x 4.6 mm, 5 μm). The gradient elution mode was selected where mobile phase-A consist of Buffer (40Mm Ammonium dihydrogen phosphate solution adjusted to pH 7.0 with orthophosphoric acid) and mobile phase-B in 60:40 ratio, whereas mobile phase B consist of solvent Acetonitrile and Methanol in the ratio 70:30. The flow rate was set to 1.5 mL/min and run time of 35 minutes. UV detector with wavelength 210 nm was used for the analysis. Column oven temperature was kept 40°C. The developed method was validated according ICH guideline and found to be linear in the range of 0.51 ppm to 1.53 ppm for N-(2-Chloroallyl)-1-(R)-aminoindan (Chloro impurity) with a correlation coefficient 0.9973. Limit of detection and limit of quantitation was found to be 0.08 ppm and 0.23 ppm respectively. Recovery for this impurity was found between 92.75% and 102.49%. Method was found to be specific, selective, precise, and robust. The developed method can successfully be applied for the determination of Chloro impurity in Rasagiline Mesylate upto very low trace level concnetration.

KEYWORDS: Rasagiline Mesylate, Genotoxic, Chloro Impurity, Development and Validation.