A simple isocratic RP-HPLC method was developed and validated for the determination of pirfenidone in bulk drug. The method was developed using C18 column with 0.5% triethylamine as aqueous phase (pH adjusted to 4.5 with orthophoshoric acid) and organic phase as acetonitrile: methanol (90:10) as mobile phase (55:45). The flow rate was maintained at 1 ml/min and detected at a wavelength of 315 nm using PDA detector. Pirfenidone was completely resolved and the retention time of pirfenidone was found to be 3.3 min. Regression analysis of the calibration data for pirfenidone showed that the dependent variable (peak area) and the independent variable (concentration) were represented by the equation Y=29654x + (-8619.9). The correlation of coefficient (R2) obtained for pirfenidone was 0.9998. Thus a good linear relationship was observed in the concentration range of 10 to 50 µg/ml of pirfenidone. The good percentage recovery clearly confirmed the reproducibility and accuracy of the developed method. The %RSD value for precision was also within the acceptable limit. The robustness studies were performed by changing the pH, wavelength and flow rate by chemometric method. The robustness factors were evaluated with the aid of full factorial design and the optimum chromatographic conditions were established by central composite design (CCD). Satisfactory data was obtained for all the method validation parameters tested. Stress degradation studies performed for pirfenidone by acid and base hydrolysis, oxidative degradation, dry heat degradation and photolysis, complied the ICH guidelines. The results of the study showed that the developed RP-HPLC method was found to be simple, rapid, precise, accurate, robust and stability indicating which can be used for routine analysis of pirfenidone.
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