Analytical RP-HPLC Method for Development andValidation of Pregabalin and Methylcobalamine inCombined Capsule Formulation

This work is concerned with application of simple, accurate, precise and highly selective reverse phase high performance liquid chromatographic (RP-HPLC) method for determination of pregabalin and methylcobalamine combined in capsule dosage form. Chromatographic separation was achieved isocratically using Waters allaiance 2695 seperation module,CI8 column (250 x 4.6 mm,5 mcg/ml) at temperature 40°C. Flow rate selected was 1 ml/min using UV visible PDA detector at 210nm. Mobile phase was prepared using ammonium dihydrogen-o-phosphate (buffer 6.0), acetonitrile and methanol in the ratio of 75:15:10 which gave better resolution and sensitivity. Developed method was validated in terms of linearity, range, specificity, precession, accuracy, robustness and ruggedness. The limit of detection was found to be 47.93mcg/ml for pregabalin and 1.33 mcg/ml for methycobalamine respectively. The limit of quantitation was found to be 145 mcg/ml and 4.05 mcg/ml for pregabalin and methycobalamine respectively. The method was found to be linear in the range of 3200-4800 mcg/ml and 16-24 mcg/ml for pregabalin and methylcobalamine respectively with correlation coefficients of 0.9938 and 0.9959. Content of pregabalin and methycobalamine was found to be 99.17 and 100.15% respectively. The validation of proposed method was verified by recovery studies and was found to be satisfactory.