Abstract

RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE DETERMINATION OF BACLOFEN IN INJECTIONS

G. Sowjanya*, D. Gowri Sankar and J. V. L. N. Seshagiri Rao

ABSTRACT

A rapid, sensitive and economical RP-HPLC method has been developed and validated for quantification of baclofen in injections. A binary mixture of potassium dihydrogen phosphate, pentane 1-sulfonic acid sodium salt buffer (pH 3.0) and a mixture of acetonitrile, methanol in the optimized gradient mode was proved to be the most suitable mobile phase for the separation of baclofen on a non-polar Symmetry C18 column. The retention time for baclofen was at 5.538 ± 0.49 min. injected at a flow rate of 0.8 mL/min. and the detection wave length was fixed at 225 nm. The method obeyed linearity in the range of 0.0050 – 0.0150 mg/mL. The % RSD was found to be less than 2% in intra-day (0.5) and inter-day (0.1) variation studies indicating that the method is precise. The recovery of baclofen was found to be in the range of 98.6 - 100.5 % indicating that the method is accurate. The method also tolerated minor variations in chromatographic conditions indicating good robustness (% RSD < 1.0). The lowest values of LOD (4.46 × 10-5 mg/mL) and LOQ (1.35 × 10-4 mg/mL) as obtained by the proposed method indicate the sensitivity of the method. The marketed injection was found to contain an average of 101.45 ± 0.05 % w/v of baclofen as stated on the label claim and the excipients did not pose any interference at the retention time of the drug indicating specificity of the method. Experiments to study the effects of forced degradation on baclofen in injections were conducted in the validation study as per ICH guidelines. This method can be used for the regular quality control analysis of baclofen in API and injections.

Keywords: Baclofen, RP-HPLC, Gradient, Symmetry C18 column, Validation.