ABSTRACT:
A new rapid, accurate, precise and economical reverse phase high performance liquid chromatographic method has been developed and validated for simultaneous estimation of Methylcobalamin, Epalrestat and Pregabalin in bulk and pharmaceutical dosage form. The separation was accomplished utilizing Agilent C18, 150 x 4.6mm, 5? column at a detection wavelength of 210nm utilizing the mobile phase water and acetonitrile 60: 40 v/v at the flow rate of 0.8ml/min and injection volume of 10µl. The total run time was 6.0?min. Validation discovered the method was specific, rapid, accurate, precise, reliable and reproducible. The calibration curve was linear over the concentration range of 0.37 – 2.25µg/ml of Methylcobalamin, 37.5-225µg/ml of Epalrestat and 37.5-225µg/ml of Pregabalin respectively with correlation coefficient of 0.999. The accuracy was determined by recovery studies and was found to be 99.5-100%. The precision of the results stated that the %RSD was <2%. The limits of detection for Methylcobalamin, Epalrestat and Pregabalin were 0.2, 0.9 and 1.2µg/ml, while the limits of quantification were 0.5, 1.5 and 0.9µg/ml respectively. Forced degradation study was carried out under acidic, alkaline, oxidative, photolytic and thermal conditions to prove the stability-indicating ability of the developed HPLC method. The high recovery confirms the suitability of developed method and can be further used in routine analysis.