Stability indicating RP-HPLC method development and validation of Etizolam and Propranolol hydrochloride in pharmaceutical dosage form

Abstract

A stability indicating RP-HPLC method was developed and validated for Etizolam and Propranolol Hydrochloride in pharmaceutical dosage form. The chromatographic separation was carried out by using PURITUS C₁₈ column (250 mm x 4.6 mm, 5 µm) with gradient mobile phase Ammoni­­um phosphate buffer pH 3 and Acetonitrile by PDA detector. Drug and degradant were monitored at 245nm, the flow rate was 1 ml/min, injection volume was 20 μl and temperature of column was 25 °C. Retention time of Propranolol Hydrochloride and Etizolam was about 7.4 min and 14.4 min respectively. Stability indicating method was performed by using 1N HCl, 1N NaOH, 3 % H₂O_2. Thermal degradation was performed at 100 °C for 7 days. The photolytic degradation was performed for 7 days. The developed method was validated as per ICH guideline using various validation parameters. Linearity was observed in concentration range 5 to 15 μg/ml of Etizolam and 200 to 600 μg/ml of Propranolol Hydrochloride with correlation coefficient of 0.9995 and 0.9983 respectively. All the parameters were within specifications. So, the developed method was found to be stability indicating, specific, linear, accurate, precise and robust.