AJAC  Vol.6 No.13 , December 2015
Development and Substantiation of a RP-HPLC Method for Monitoring of Impurities in Pirfenidone Drug Substance
Abstract: A simple, rapid and rugged RP-HPLC method was developed for evaluation and quantification of impurities present in Pirfenidone (PFD) drug substance. Impurities were separated and determined on a Zorbax RX-C18 column (250 mm length, 4.6 mm inner diameter and 5.0 μm particle size, octadecylsilane chemically bonded to porous silica) with 0.02 M KH2PO4 buffer and acetonitrile as mobile phase using a simple gradientelution program. The column flow rate of 1.0 mL per minute was used for the separation. The detection wave length was fixed at 220 nm. The method was substantiated with respect to specificity, precision, linearity, range, accuracy, ruggedness, limit of detection and quantitation. The impurities were identified as 2-hydroxy-5-methylpyridine and Iodobenzene. The linearity range obtained was 0.017 to 0.380 μg/mL for 2-hydroxy-5-methylpyridine, 0.047 to 0.382 μg/mL for Pirfenidone and 0.030 to 0.99 μg/mL for Iodobenzene with the retention times of 3.248 min, 10.608 min and 24.241 min for 2-hydroxy-5-methylpyridine, Pirfenidone and Iodobenzene, respectively. The percentage recoveries of 2-hydroxy-5-methylpyridine and Iodobenzene were in the range of 94.08% - 104.12%. The LOD and LOQ values were found 0.000005 mg/mL, 0.000017 mg/mL for 2-hydroxy-5-methylpyridine and 0.009 μg/mL, 0.030 μg/mL for Iodobenzene, respectively. The method is found to be suitable for the quantitation of impurities along with Pirfenidone drug substance. The method was validated as per the International Conference on Harmonization (ICH) guidelines.
Cite this paper: Bodempudi, S. , Babur, R. and Reddy, K. (2015) Development and Substantiation of a RP-HPLC Method for Monitoring of Impurities in Pirfenidone Drug Substance. American Journal of Analytical Chemistry, 6, 1019-1029. doi: 10.4236/ajac.2015.613097.

[1]   Ministry of Health, Labor and Welfare (2008) Report on Deliberation Results. Nonproprietary Name: Pirfenidone Evaluation and Licensing Division, Pharmaceutical and Food Safety Bureau.

[2]   (2010) CHMP Assessment Report. International Nonproprietary Name: Pirfenidone Procedure No. EMEA/H/C/002154. European Medicines Agency.

[3]   Raghu, G., Collard, H.R., Egan, J.J., Martinez, F.J., Behr, J., Brown, K.K., et al. (2011) An Official ATS/ERS/JRS/ ALAT Statement: Idiopathic Pulmonary Fibrosis: Evidence-Based Guidelines for Diagnosis and Management. American Journal of Respiratory and Critical Care Medicine, 183, 788-824.

[4]   Tong, S., Wang, X., Jiang, H., Xuegu, X., Pan, Y., Kunming, C., et al. (2010) Determination of Pirfenidone in Rat Plasma by LC-MS-MS and Its Application to a Pharmacokinetic Study. Chromatographia, 71, 709-713.

[5]   Shi, S., Wu, J., Shi, S., Wu, J. and Zeng, F. (2008) Development and Validation of an Improved LC Method for the Simultaneous Determination of Pirfenidone and Its Carboxylic Acid Metabolite in Human Plasma. Chromatographia, 69, 459-463.

[6]   Wang, Y., Zhao, X., Zhong, J., Chen, Y., Liu, X. and Wang, G. (2006) Simple Determination of Pirfenidone in Rat Plasma via High-Performance Liquid Chromatography. Biomedical Chromatography, 20, 1375-1379.

[7]   Tamilselvi, N. and Krurian, D.S. (2012) Bioanalytical Method Development and Validation of Pirfenidone by RP-HPLC Method and Its Application to the Determination of Drug Food Interaction Study in Wister Rats. International Journal of Pharmaceutical and Biomedical Research, 3, 132-142.

[8]   (2005) International Conference on Harmonization. Guidance on Validation of Analytical Procedure: Text and Methodology. ICH-Q2 (R1). IFPMA, Geneva.

[9]   ICH Stability (2003) Testing of New Drug Substances and Products Q1A (R2). International Conference on Harmonization, IFPMA, Geneva.

[10]   ICH Photo Stability (1996) Testing of New Drug Substances and Products Q1b. International Conference on Harmonization, IFPMA, Geneva.