Stability indicating method development and validation for determination of Pirfenidone in raw material by reversed-phase high performance liquid chromatography.

摘要

Idiopathic pulmonary fibrosis (IPF) is a fatal disease of unknown cause where the lung tissues become damaged and patients experience shortness of breath, cough and ultimately death. There is a great interest in drugs used to manage this deadly disease.;At present, the only approved medical treatment for IPF is Pirfenidone. It is a multifunctional, orally available small molecule with anti-fibrotic, anti-inflammatory, and anti-oxidative activities. It is effective for the treatment of patients with mild-to-moderate IPF.;A reversed-phase liquid chromatography method was developed on an Agilent Zorbax SB C18 column (250 x 4.6 mm, 5 mum) to analyze Pirfenidone in raw material and determine impurities/degradants. The mobile phase composition was optimized to be 27% ACN and 73% buffer. The buffer that gave the most suitable results was monobasic potassium phosphate buffer set at a pH of 2.9. The injection volume was set at 10 muL. The flow rate was set at 1.0 mL/min. The UV absorption wavelength was set at 316 nm. The total run time of the developed method was 16 minutes.;Degradation studies were carried out under different stress conditions: acid hydrolysis, base hydrolysis, oxidation, heat, and UV light. The developed method was validated based on the ICH and FDA guidelines by performing system suitability, solution stability, specificity, method robustness, limit of detection, limit of quantitation, linearity, accuracy, and precision studies.