A comprehensive review on analytical and bioanalytical methods of molnupiravir

Molnupiravir (MLP), an orally bioavailable prodrug of N4hydroxycytidine (NHC), advanced rapidly during the COVID19 pandemic as a broadspectrum antiviral drug.The COVID-19 pandemic has claimed a considerable number of lives. This review consolidates MLP’s physicochemical characteristics, mechanism and pharmacokinetics to guide analytical targeting particularly the predominance of circulating N-hydroxy cytidine (NHC) then surveys spectrophotometric, spectrofluorimetric, chromatographic (HPLC, UPLC, HPTLC), bioanalytical LC–MS/MS, and capillary zone electrophoresis (CZE) methods for MLP/NHC and degradants in pharmaceutical and biological matrices. Spectrophotometric sections detail rapid UV assays using greener solvents (water, ethanol, micellar systems) with validation parameters. Spectrofluorimetry highlights ng mL− 1 sensitivity via Zn(II) chelation and notes interpretive care for nanomaterial based probes. Chromatography covers isocratic RP HPLC/UV on C18 columns for routine QC, stability indicating methods with forced degradation profiling, eco designed micellar HPLC and DoE optimized methods, HPTLC for high throughput content uniformity and coformulation separations with favipiravir. Bioanalytical LC–MS/MS focuses on NHC and MLP in plasma and saliva using protein precipitation or SPE, C18 or polarembedded phases, and SIM/MRM detection across wide linear ranges. Capillary zone electrophoresis (CZE) and derivative ratio spectrophotometry offer solvent lean, cost effective dual drug assays. The antiviral medications under study were measured using these established techniques in biological fluids, co-formulated synthetic mixes, and single dosages Therefore, the goal of this study is to highlight the most recent analytical techniques for the testing of drugs.