Development and validation of stability indicating RP-HPLC method for simultaneous estimation of pregabalin and aceclofenac in bulk and pharmaceutical dosage form

Abstract

A simple, precise and reproducible Reverse Phase High Performance Liquid Chromatography method was developed and validated for simultaneous estimation of Pregabalin and Aceclofenac in tablet dosage form. Chromatographic separation was achieved by Grace C₁₈ (250 mm x 4.6 ID , Particle size- 5 micron) column and methanol : water pH3 (60:40v/v) as mobile phase, at a flow rate of 1 ml/min (millilitre per minute) using UV detection at 216nm. Forced degradation experiments were carried out by exposing Aceclofenac and Pregabalin standard and sample for thermal, photolytic, oxidative and acid-base hydrolytic stress conditions. The method has been validated for linearity, accuracy, precision, LOD, and LOQ. The retention time for Aceclofenac and Pregabalin were obtained as 4.296 min and 5.955 min respectively. Linearity of Aceclofenac and Pregabalin were found to be in range 20-100μg/ml and7.5-37.5μg/ml.(R²=0.998) respectively. The accuracy of present method was evaluated at 50%,100%,150%. Recovery was found to be in a range from 99.80%-100.42% for both of the drugs. Intermediate precision studies were carried out and the RSD values were less than 2%. Lower values of LOD (0.35µg/ml for ACF and 0.18 µg/ml for PRE) and LOQ (1.08µg/ml for ACF and 0.56 µg/ml for PRE) indicated good sensitivity of the method. In this study, the optimization of mobile phase, flow rate, injection volume and wavelength were achieved. This demonstrate that the developed method is simple, precise, accurate and robust for simultaneous estimation of aceclofenac and pregabalin in tablet dosage form. The method was acceptable for degradation studies of heat, sunlight, acid, base, peroxide which meet the acceptance criteria for forced degradation studies.