Two steps non-enzymatic synthesis of molnupiravir, which is free from mutagenic impurity and analytical method development for estimation of genotoxic impurity (Hydroxylamine hydrochloride content) by using RP-HPLC technique

Genotoxic impurity free Molnupiravir was synthesised by a novel and an elegant non-enzymatic method, which is simple operationally and ecologically. The genotoxic raw material, Hydroxylamine, was used in the initial stages of synthesis to get the mutagenic impurity free Molnupiravir. A new and sensitive HPLC method was developed and validated for the determination of hydroxylamine in Molnupiravir drug substance according to ICH guidelines. The HPLC method was developed and optimized on Zorbax SB C18 150×4.6mm, 3.5μm column with oven temperature maintained at 40°C. 1.0 mL of Orthophosphoric acid in 1000 mL water was selected as mobile phase in isocratic reverse phase mode. Chromatographic parameters are flow rate: 0.8 ml/min, wavelength detection: 252 nm, injection volume: 10μl and run time: 20 min. Based on the validation data; the method was found to be specific, sensitive, accurate and precise. This method can be used as a good quality control tool for quantification of hydroxylamine at the low level. The experimental data was deliberated in detail in this research paper.