RP-HPLC Method development and Validation for Rapid estimation of Diazepam in Bulk and Pharmaceutical Dosage Form

Abstract

The main objective of the study is to establish simple, precise and rapid RP-HPLC method for determination of diazepam and its decomposition products present in bulk and pharmaceutical dosage form. Various parameters that can potentially affect the analytical method were investigated and optimized. The compounds were well resolved in an isocratic method using the mobile phase composition of potassium dihydrogen phosphate buffer(0.05M) along with diluted o-phosphoric acid adjusted to pH 4.5: methanol (40:60v/v) at a flow rate of 1 ml/min using C18 (25cm x 0.46 cm) Hypersil BDS column. The detection was carried out at wavelength of 248nm.The retention time of diazepam was 3.3 min. The developed method was validated by evaluating various parameters such as linearity, precision, accuracy, robustness, specificity, limit of detection, and limit of quantification according to the international council for harmonization guidelines. The standard calibration curve was obtained in the concentration range of 20µg/ml for diazepam. The % of label claim was found out to be 93.66 for diazepam. Observation of all these parameters leads to the point that developed RP-HPLC method is accurate, precise, linear, specific and robust. It can be successfully adopted for routine quality control analysis of Diazepam in bulk and pharmaceutical dosage form without any interference.