Development and Validation of Analytical Method for Simultaneous Estimation of Diclofenac sodium and Tramadol HCl in Bulk and its Formulation

Abstract

A simple, accurate and precise UV Spectrophotometric method and stability indicating RP- HPLC method was developed and validated for simultaneous estimation of Diclofenac sodium and Tramadol HCl in bulk and its formulation. UV first order derivative spectrophotometric method based on the measurement of absorbance at 248.43 ZCP of Diclofenac sodium and at 270.86 nm ZCP of Tramadol HCl. The calibration curve was linear in a concentration range of 15-52.5 μg/ml for Diclofenac sodium and 10-35 μg/ml for Tramadol HCl. Correlation coefficient was 0.9991 and 0.9987 for Tramadol HCl and Diclofenac sodium, respectively. The RPHPLC method has shown adequate separation of Diclofenac sodium and Tramadol HCl in bulk and its Formulation. The separation was achieved on BDS Hypersil C18, 250mm × 4.6mm, 5μ(particle size), with an isocratic mixture of Methanol: 50mM phosphate buffer pH 3 adjusted with ortho-phosphoric acid in the ratio of 65 : 35 v/v. The mobile phase at a flow rate of 1.0 ml/min, Injection volume 20μl and wavelength of detection was kept at 270 nm. The retention time of Tramadol HCl and Diclofenac sodium was 1.703 ± 0.1min and 7.837 ± 0.1min, respectively. The Tramadol HCl and Diclofenac sodium were undergoes degradation under acidic, basic, oxidative and thermal conditions. The Method was linear over in the concentration range of 20- 32.5μg/ml and 30-48.75μg/ml, Correlation coefficient was 0.999 and 0.998 for Tramadol HCl and Diclofenac sodium, respectively. The developed method was validated as per ICH guideline, for its accuracy, precision, LOD & LOQ and the results were found to be satisfactory thus the method is specific, rapid and simple with good sensitivity for estimation of Tramadol HCl and Diclofenac sodium. The developed method can be used successfully for routine analysis of the drug in bulk and its formulation.