Validation of Stability Indicating RP-HPLC Method for the Simultaneous Estimation of Pregabalin and Methylcobalamin in Formulation by QBD Approach

Aasadi Balaji*, G. Dharmamoorthy, G. Aruna, KV Nanda Kumar, P. Jaya Chandra Reddy
Krishna Teja Pharmacy College, Chadalawada Nagar, Renigunta Road Tirupati-517506

ABSTRACT
By considering the current regulatory requirement for an analytical method development, a simple, precise, accurate, linear and rapid Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method for the simultaneous quantitative estimation of Pregabalin 75mg and Methylocobalamin 750µg in capsule has developed using analytical quality by design approach.The optimized method was achieved using Unisol C-18 (3μm, 4.6 ×150mm) column with mobile phase consisting of mixture of water and methanol (40:60v/v) with a flow rate of 0.6ml/min at 210nm. The developed method resulted in pregabalin eluting at 5.540min and methylcobalamin eluting at 2.593min. The method exhibited linearity over the range of 50-150µg/ml and 5-15µg/ml for pregabalin and methylcobalamin respectively. The method precision is exemplified by %RSD of 0.999 for both pregabalin and methylcobalamin. Accuracy studies revealed % mean recoveries during spiking experiments between 98 and 102. The limit of detection was obtained as 2.69µg/ml for pregabalin and 1.74µg/ml for methylcobalamin, while the limit of quantitation was obtained was 8.18µg/ml for pregabalin and 5.27µg/ml for methylcobalamin. There are no interfering peaks underperformed degradation conditions. Therefore, a simple, precise, accurate, linear and rapid RP-HPLC method was developed and validated as per ICH guidelines and hence can be applicable in routine analysis for tablets in various pharmaceutical industries.
Keywords:  Pregabalin, Methylcobalamin, QbD, RP-HPLC, Validation.