Development and validation of a HPLC method for the assay of dapivirine in cell-based and tissue permeability experiments

► A HPLC-UV method was developed for assaying dapivirine in biological matrices. ► Box-Behnken experimental design was successfully used to optimize the method. ► Validation according to FDA and EMA guidelines on bioanalytical assays was performed. ► Method was applied to cell-based/mucosal tissues uptake and permeability assays.