Display omitted •Novel green HPTLC method for simultaneous quantitation of sofosbuvir and ledipasvir.•Forced degradation stability study with HPTLC-ESI-MS detection of the products.•Structure elucidation of each fragment in the mass spectra and fragmentation pattern.•Eco-Scale greenness assessment with a 93 score (excellent green)•Method validation and application on synthetic mixtures and pharmaceutical tablets. Recently, the analytical community has shown increasing interest in developing more environment-friendly practices by eliminating or reducing the use of hazardous chemicals and solvents which are extremely dangerous to human health and the environment. Thus in this study, a simple, rapid, inexpensive and eco-friendly HPTLC method with densitometric detection was designed, optimized and validated for the simultaneous quantitation of the antiviral drugs sofosbuvir (SOF) and ledipasvir (LED) in pure powder, synthetic mixtures, combined pharmaceutical formulation and in the presence of their stressed degradation products with high sensitivity and no interference. The separation was carried out on silica gel F254 plates, using green mobile phase ethyl acetate: water: ethanol (94:5:1v/v/v) for development followed by densitometric detection of SOF and LED at 250 and 320 nm, respectively. Both drugs had second order polynomial calibration curves within ranges 100–5000 ng/spot and 100–3000 ng/spot for SOF and LED, respectively, with correlation coefficients >0.9998 and the limits of detection were 30 and 29 ng/spot and the limits of quantification were 90 and 88 ng/spot for SOF and LED, respectively. The method was validated in terms of linearity, accuracy, precision, selectivity, and robustness. The drugs were subjected to acidic and basic hydrolysis, oxidation and photolytic conditions as per ICH guidelines, followed by detection of the degradation products and structure elucidation of their fragments with the aid of the HPTLC-ESI- MS technique. The greenness of the proposed method was assessed using the analytical Eco-Scale as an assessment tool with a 93-point score. So, it is considered excellent green.