Analytical Method Development and Validation for Pirfenidone (Anti-Fibrotic) in bulk drug and dosage form

Katikala Dharmendra*, Dr. C. Sreedhar, T. Sreenivasa Rao, Rikshaya Tamang, Meghnath Chaudhary, Mohamed adam
Department of Pharmaceutical Analysis, Karnataka College of Pharmacy, #33/2,Thirumenahalli,Hegde Nagar main road, Bangalore – 560064, KA. India.

A B S T R A C T
Pirfenidone, 5-methyl-1-phenyl-2-(1H)-pyridine is a novel Anti-fibrotic agent approved for mild to moderate Idiopathic pulmonary fibrosis (IPF).  Isocratic reverse phase high performance liquid chromatography (RP-HPLC) separation using a XBridge C18 column of particle size 5µm, (4.6×250mm). The separations were achieved at the UV detection at 311nm using the mobile phase of Acetonitrile: Trifluoro acetic acid in the ratio of 50:50,with apparent pH of 2.08,the Flow rate was 0.8ml/min and the injection volume was set at 20 µl with 10mins of runtime. The retention time was observed at 4.617mins for Pirfenidone. The method was validated by using various validation parameters like accuracy, precision, linearity, limit of detection (LOD), limit of quantification (LOQ). The standard curve was linear over a working range of 1- 32 µg/ml and gave an average correlation factor of 0.9988 for Pirfenidone. The limit of detection and the limit of quantification were found to be 0.120µg/ml and 0.401 µg/ml for Pirfenidone respectively. The method showed good recoveries and relative standard deviations of intra and inter day assay less than 2. This method can be easily and conveniently used for routine analysis of Pirfenidone in bulk Drug and tablet dosage forms.
Keywords: Pirfenidone, RP-HPLC, UV–Visible Spectrophotometer, LOD, LOQ.