Development and validation of RP-HPLC method for simultaneous estimation of propranolol and clonazepam
Keywords:RP-HPLC, Propranolol, Clonazepam, Simultaneous estimation, Validation.
A reversed phase high performance liquid chromatographic method for routine analysis of propranolol and clonazepam has been developed using Discovery C18 column (4.6X150mm, 5µm) dimensions at ambient temperature .The mobile phase used for this study was mixture of potassium hydrogen phosphate and Acetonitrile at 45:55 ratio. The Mobile phase was pumped from the solvent reservoir to the column at a flow rate of 1 ml/min for 6min. The elution was monitored at 238nm. Propranolol and Clonazepam were eluted at 2.191 min and 2.935 min respectively with good resolution. The optimized method was then validated according to the ICH guidelines. %RSD of the Propranolol and Clonazepam were and found to be 0.5 and 0.4 respectively. %Recovery was obtained as 99.53% and 99.38% for Propranolol and Clonazepam respectively. LOD, LOQ values obtained from regression equations of Propranolol and Clonazepam were 0.18, 0.54 and 0.004, 0.014 respectively. Therefore, a sensitive, robust, accurate was developed.