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.

Development and validation an HPLC - UV method for determination of esomeprazole and pirfenidone simultaneously in rat plasma: application to a drug monitoring study

“…The method was found linear (r 2 =0.9985) between 47 and 382 ng/mL. LOD was found 14 ng/mL and LOQ was calculated as 9.4 μg/ mL (Bodempudi et al, 2015).…”

“…In Bodempudi et al (2015) study, an HPLC based chromatographic method was developed for the determination of pirfenidone in the drug substance. Separation was achieved with a C18 column (4.6 mm x 250 mm i.d., 5 μm p.s.)…”

“…Also, some analytical methods were reported for the determination of pirfenidone from the different matrix which includes pharmaceutical dosage forms and plasma. These are spectrophotometric (Thorat, Padmane, Tajne, & Ittadwar, 2016), spectrofluorometric (Sambhani & Biju, 2018), capillary electrophoresis (Sotgia et al, 2020), high-performance thin layer chromatography (Thorat et al, 2016), gas chromatography (Ma et al, 2017), and HPLC with an ultraviolet detector (Bodempudi, Babur, & Reddy, 2015;Parmar et al, 2014;Ravisankar, Anusha Rani, Devadasu, & Devala Rao, 2014;Thorat et al, 2016), a photodiode array detector (Bodempudi et al, 2015) and a mass spectrometry detector (Tong et al, 2010;Wen et al, 2014) methods.…”

Development of a Topical 48-H Release Formulation as an Anti-scarring Treatment for Deep Partial-Thickness Burns

“…PF was determined from a standard calibration curve prepared starting from a stock solution containing 1 mg/ml PF in 5 ml MeOH (Fig. 2) in the same manner as that for the UV technique (18,19).…”

A QbD‐led simple and sensitive RP‐UHPLC method for simultaneous determination of moxifloxacin, voriconazole, and pirfenidone: An application to pharmaceutical analysis

“…Most articles report the method development of individual analytes or in combination with analogous drugs within the same family. However, these reports state the use of higher concentrations of buffer modifiers with a need for pH adjustment, which is a technique with independent variables, high organic content, poor sensitivity and longer acquisition time and is uneconomical (Bodempudi et al, 2015; Czyrski et al, 2017; Kaur et al, 2021; Yıldırım et al, 2020). Collectively, there is a need to eradicate the disadvantages of the existing approaches by inoculating a novel method to overcome the failures of the existing ones.…”