Assay method development and validation for butamben drug substance by using high performance liquid chromatographic technique

Abstract

Stability indicating assay methodology was developed with UV detection for the simultaneous quantification of Butamben drug substance by HPLC. The separation between degradation impurities and the peak due to Butamben was achieved by using Universal C 18, (100 x 4.6) mm, 5.0-micron column. Mobile Phase was made up of Water, Methanol and Orthrophosphoric acid by mixing 350:650:1 ml, respectively. Isocratic method was used by using flow rate of 1.0 ml/min, UV at 250 nm and column oven temperature at 50°C. Relative standard deviation for standard preparation under system precision was achieved within acceptance criteria. Relative standard deviation for retention time was achieved 0.11%, which shows reproducibility during replicate injections. After the successful development of this method, chemically forced degradation was performed by external acid, alkali and peroxide treatment. Not any degradation peak was interfering with any impurity. This newly developed innovative method is found suitable for the Assay analysis of Butamben API.