DEVELOPMENT AND VALIDATION OF STABILITY INDICATING HPLC METHOD FOR ESTIMATION OF RELATED SUBSTANCES IN AVANAFIL

The current proposal of the research is to estimation of related substances by using high-performance liquid chromatographic method has been developed and validated for the determination of Avanafil. Reversed-phase chromatography was performed on Waters 2489 UV 2695 pump, Waters 2998 PDA 2695 pump Software Empower2 photodiode array detector using ACE C18 (250 mm × 4.6 mm, 5 μm particle size) column with pH 4.2 buffer: methanol in the ratio of 90:10 as mobile phase A and acetonitrile as mobile phase B at a flow rate of 1.0 mL/min. by gradient elution with UV detection at 245 nm. Recovery and Linearity was observed well within the limits (R2 = more than 0.99 for concentration range of LOQ to 150% level for linearity and the % recovery was within the ICH acceptance limits of 85-115% ) for all the impurities. The limit of quantitation (LOQ) and limit of detection (LOD) were found to be less than 0.05%. The method was validated as per ICH guidelines. The RSD for intra-day and inter-day (<3.0% RSD) precision were found to be less than 1 %. The percentage recovery was in good agreement with the labeled amount in the pharmaceutical formulations. from the method validation data, it can be concluded that the method is simple, specific, precise and accurate for the determination of Avanafil in pharmaceutical formulations.