Method Development and Validation for Related Impurities of Efavirenz by RP-HPLC Method

Abstract

A simple, specific, validated reverse phase high performance liquid chromatographic method has been developed for the quantification of related impurities of Efavirenz in bulk drug substance. In this method impurities are quantified successfully at 0.2% quantification threshold. This method utilizes high purity silica based RP-C18 column i.e Inertsil-ODS 3 V (250 × 4.6 mm × 5 μm) with which efficient separation and resolution was achieved at a temperature of 45°C with a flow rate of 1.5 ml/min. 0.1% o-Phoshoric acid and Acetonitrile in the ratio of 20:80 v/v was employed as a mobile phase and samples were scanned at a wavelength of 245 nm. Validation of the method was successfully established by performing various validation parameters such as specificity, precision, linearity, accuracy, LOD, LOQ, robustness, ruggedness according to ICH guidelines. A strong linear relation exists over the range of 20 to 120% specification level with a correlation coefficient value of 0.999. The method was sufficiently accurate, precise, robust and rugged with a good recovery, acceptable standard deviation and system suitability parameters within the stipulated limits. The method sensitivity was accomplished by LOD & LOQ. This method can be successfully employed for the routine in process quality control and for the quantification of impurities in bulk drug substance.