A VALIDATED STABILITY INDICATING METHOD FOR THE DETERMINATION OF RELATED SUBSTANCES IN BELINOSTAT AND ITS DEGRADATION IMPURITIES BY RP-HPLC

Krishna Katta, Tirukkovalluri Siva Rao*, J. Mosesbabu and D. Venugopala Rao

ABSTRACT

A new sensitive, robust and stability indicating RP-HPLC method was developed for quantitative determination of related impurities and its degradation impurities in belinostat drug substance. The excellent chromatographic resolution was achieved on a Kromasil C18 (250 mm×4.6 mm, 4.0 μm) column with a mobile phase containing 0.1% perchloric acid in gradient combination with acetonitrile (ACN) at a flow rate of 1.0 mL/min and the eluent was monitored at 210 nm. The developed method is capable to separate all six known impurities and degradation impurities from Belinostat with a resolution (Rs) greater than 2.0. Performed method validation according to ICH guidelines, achieved the % RSD <0.2% in precision study. The Limit of detection and quantification for related impurities were 0.017 and 0.050 μg/mL respectively. Regrreation analysis proved that the method is linear with an r2 value (correlation coefficient) greater than 0.999 for all related impurities. The method is found to be accurate with % recoveries ranging from 98 to 100. Established homogeneity of belinostat peak and proved the stability indicating nature of the method by conducting forced degradation studies under elevated temperature, photolysis, Acid hydrolysis, Base hydrolysis, water hydrolysis, and oxidation.

Keywords: Belinostat; Validation; Impurities and Degradation Products.