Ecofriendly stability-indicating UHPLC-PDA method for determination of the new influenza antiviral prodrug Baloxavir Marboxil; application to degradation kinetic studies and structure elucidation of the major degradation products using LC-MS

Abstract

Implementation of green chemistry principles in analytical procedures is of great importance. Green analytical chemistry minimizes the dangerous impact on human beings and the environment that is the primary objective nowadays. In the proposed work, a well-developed, fully validated and ecofriendly stability indicating UHPLC coupled with photodiode array detector (PDA) method was established for the determination of Baloxavir Marboxil (BXM), a novel antiviral drug for the treatment of influenza A and B, in raw materials and pharmaceutical formulations. The method employed Inertsil ODS-3 C18 column to separate BXM from its forced degradation products using a mobile phase composed of 0.1% citric acid (pH 2.5 ± 0.05)-methanol (50:50, v/v), at a flow rate of 0.8 mL/min in an isocratic separation mode. BXM was monitored using diode array detector at 260.0 nm. BXM stability was inspected using various stress conditions as per ICH recommendations. The impact of acid, base hydrolysis, and oxidative stress conditions was greater on BXM in comparison to photo and thermal hydrolysis. The formed degradation products were injected into the mass spectrometer (MS) to elucidate their structures. The suggested pathways for degradation mechanisms were presented. Moreover, the proposed method was effectively utilized to study the degradation kinetics of acid, base and oxidative degradation of BXM. Additionally, four assessment tools were employed to evaluate the greenness of the developed method, namely; the National Environmental Method Index, the Analytical Eco-scale, Green Analytical Procedure tool, and the Analytical Greenness metric. Moreover, a comparative study of the method's greenness was conducted with the reported HPLC method and the results confirming that the suggested method is superior regarding the guidelines and principles of green chemistry. In addition to, having higher qualitative and quantitative scores in comparison to the reported method, and providing reliable and precise results of the method greenness.