Development and Validation of RP-HPLC Method for Stress Degradation Studies of Pregabalin

Abstract

A simple and selective stability indicating Reverse-Phase High Performance Liquid Chromatographic (RP-HPLC) method has been developed and validated for estimation of pregabalin. The separation of all degradation peaks was achieved on 250 mm × 4.6 mm, 5μ, Alltima, C18 column, using gradient elution of Phosphate buffer : Acetonitrile (96:4)(Mobile phase A) and Acetonitrile (Mobile phase B) at flow rate of 1.0 mL/min and UV detection at 210 nm with the total run time of 75 min. Retention time of pregabalin was found to be 9.9 min. Forced degradation of pregabalin was carried out under acidic, basic, oxidative, thermal and humidity conditions. It was found that pregabalin degrades under basic and oxidative conditions. The method was validated for system suitability, specificity, linearity, accuracy, precision, LOD, LOQ and robustness as per ICH guidelines. Compatibility study of pregabalin was carried out with lactose. Pregabalin was reacted with lactose to generate lactose conjugate. It was found that there is decrease in peak area of pregabalin and lactum conjugate elutes at 46.02 min which very well coincides with the related literature. The method was found to be specific enough to separate degradation products from main analytes. Therefore the developed RP-HPLC method can be used for the estimation of pregabalin.