ArticleRP-HPLC METHOD DEVELOPMENT, VALIDATION AND STABILITY INDICATING STUDIES FOR THE ESTIMATION OF AMITRIPTYLINE HYDROCHLORIDE AND PERPHENAZINE IN BULK
D. Titus , Dr. K. Srinivas Reddy , SK. Nasreen , Dr. D. Kumara Swamy , CH. Sumanth , Dr. K. Praveen Kumar
A new, simple, precise, accurate and reproducible RP-HPLC method was validated for the estimation of Amitriptyline Hydrochloride (AMT) and Perphenazine (PRP) & Stability indicating Studies were performed. Separation of Amitriptyline Hydrochloride and Perphenazine was successfully achieved on Hypersil ODS (250x4.6mm) 5µm column in an isocratic mode utilizing Methanol:Water (90:10) at a flow rate of 1.0 ml/min and eluents were monitored at 247 nm with a retention time of 4.235 and 3.490 minutes for and Amitriptyline Hydrochloride and Perphenazine respectively. The method was validated and it was found to be linear and the range is 10-100μg/ml for both drugs. The values of the correlation coefficient were found to 0.9995 for Amitriptyline Hydrochloride and 0.9912 for Perphenazine respectively. The LOD for Amitriptyline Hydrochloride and Perphenazine were found to be 0.23µg/ml and 0.39µg/ml. The LOQ for Amitriptyline Hydrochloride and Perphenazine were found to be 0.72µg/mL and 1.18µg/mL respectively. The mean recoveries obtained were 102.3% and 100% of Amitriptyline Hydrochloride and Perphenazine, which indicates accuracy of the proposed method. Forced degradation of Amitriptyline Hydrochloride and Perphenazine in various conditions like alkaline, acidic, oxidative and thermal degradation was performed in this investigation. The content of degradation of the drugs was quantitatively analyzed by HPLC. The Amitriptyline Hydrochloride is very sensitive drug it was degraded in all conditions while Perphenazine is more stable in alkaline, acidic, and oxidative than thermal conditions. Proposed method was validated for precision, accuracy, linearity & range and robostness according to ICH guidelines. The method was successfully applied to Perphenazine and Amitriptyline Hydrochloride estimation in bulk.
Amitriptyline HCl, Perphenazine, Hypersil C18, Forced degradation, ICH guidlines.