Abstract

A NOVEL VALIDATED METHOD FOR DETERMINATION OF GENOTOXIC IMPURITES IN BEPOTASTINE BESILATE (API) WITH RESERVE PHASE HPLC TECHNICS.

R. K. Phadke* and V. D. Gaitonde

Abstract

The purpose of this research study was to develop a novel, simple, precise, accurate and robust method for determination of genotoxic impurities as Methyl benzene sulphonate and Butyl benzene sulphonate in Bepotastin Besylate API. The Chromatographic analysis was performed on Kromasil C8 column (250 × 4.6 mm, 5 μm) with Mobile phase (a) was 2.3 g of ammonium dihydrogen phosphate dissolved in 500 mL of water with adjusted the pH 3.0 ± 0.05 by dilute O-phosphoric acid and then further diluted to 1000ml with water. Filtered this solution through a 0.45μm membrane filter paper, and degas for 5 minutes by Sonication technique. The mobile phase (b) was acetonitrile. The initial linear gradient proportion of mobile phase (a) and mobile phase (b) was 70:30 then slowly increase the solvent concentration from 30 % to 70% within 12min and same has been continued for 20 min. Again reduced the concentration of solvent to achieve the initial composition within 25 min and then continued up to 35 min. The chromatographic elution was monitored by waters 2489 UV/visible detector. The peak detection was done at 215 nm wavelength. This chromatographic method allows for detection of Methyl benzene sulphonate at 0.012ppm (MBS) and Butyl benzene sulphonate (BBS) at 0.019ppm respectively, where as for quantification 0.036ppm and 0.057 ppm respectively. The linear response was found in a concentration range of 0.40-0.60 ppm with a squared correlation coefficient was 0.9998 and 0.9993. The mean recovery was found to be 96.96 and 95.36% respectively. Hence the developed method was simple, fast, linear, accurate, reproducible and robust. The same method was validated by following ICH guideline parameter.

Keywords: HPLC; Methyl benzene sulphonate; Butyl benzene sulphonate; Kromasil C8, HPLC with UV/PDA detector.