Development and Validation of Stability Indicating RP-HPLC Method for the Determination of Impurity Profile in Gamendazole: Experimental Male Oral Contraceptive

Abstract

A novel RP-HPLC method has been developed for the simultaneous determination of Gamendazole and its process impurities and degradation impurities. Chromatographic separation was achieved on a C18 column (size: 250 mm × 4.6 mm; 5 µm particle size) using a gradient program at a flow rate of 1.0 mL/min and an injection volume of 10 µL with 210 nm detection. The mobile phase was water–acetonitrile (pH of water was adjusted to 3.0 with orthophosphoric acid) through gradient elution.

The product was subject to stress conditions like acid, base, peroxide, thermal, and photolytic degradation. Similar degradants were observed in acid and base degradation. No degradants were observed in peroxide degradation. Several new degradants were observed in photolytic degradation. However, no interference was observed due to degradation impurities and Gamendazole, known impurities and unknown impurities.

As part of the method validation, specificity, limit of detection (LOD), limit of quantitation (LOQ), linearity, accuracy, precision, robustness, and ruggedness were determined. LOQ values for Gamendazole, Impurity-A, Impurity-B, and Impurity-C were 0.04%, 0.04%, 0.02%, and 0.04%, respectively. Good linearity (r² > 0.99) was obtained ranging from LOQ to 150%. Recovery was verified for all impurities at concentrations ranging from LOQ to 150%. %Recovery for Impurity-A, Impurity-B, and Impurity-C were 99.7–107.1%, 99.1–108.9%, 98.6–107.2%, respectively.

Keywords:

• degradation studies
• development
• Gamendazole
• impurity profile
• RP-HPLC
• validation

Acknowledgments

The authors would like to acknowledge Suven Life Sciences, Hyderabad, India, for supporting this work. The authors would like to express special thanks of gratitude to Mr. Ch.V.S.L. Kamewar Rao and Madhusudhan Reddy Bethi for their encouragement. The authors also would like to acknowledge Kanakadandi Rama Subhramanya Sarma for participation in preliminary laboratory work and Gogireddy Surendra Reddy for providing the samples.