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.
