Development and Validation of Stability Indicating HPLC Method for Estimation of Novel Nitroimidazooxazine Antitubercular Drug Pretomanid Followed by LC-MS

Abstract

Introduction: A reliable and fast method using reversed phase high performance liquid chromatography (RP-HPLC) has been developed and validated to determine the concentration of pretomanid in the presence of its degradation products. The drug was subjected to various stress tests according to international guidelines, and the results showed that pretomanid is susceptible to degradation under certain conditions such as alkali hydrolysis and photolysis at room temperature.

Objective: To develop a simple, precise, accurate, and economic stability indicating analytical methods for determination of Pretomanid by HPLC .To validate developed methods as per ICH guidelines. To study forced degradation study of selected drugs at recommended conditions in guidelines which are thermal, photolytic, oxidation and acid/ alkaline hydrolysis and identify the degradation products by LC-MS

Method: The RP-HPLC method was effective in separating the drug from its degradation products using a HiQ Sil C18 column and a mobile phase composed of 10 mM ammonium acetate solution pH3: methanol (65:35 v/v). The validation study demonstrated that the method was accurate, precise, and robust. LC-MS was used to identify the degradation products and to suggest possible fragmentation.

Result: Developed method was validated and found to be linear with R²= 0.9993 for concentration in the range 5-30 µg/ml, accurate, precise, robust and  Limit of detection (0.817 µg/ml) and limit of quantitation (2.476 µg/ml)

Conclusion: This method can be useful for assessing the stability of pretomanid in commercial pharmaceutical dosage forms. No previous method has been documented for the degradation behavior of pretomanid, and chemical analysis through mass spectrometry