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Daclatasvir dihydrochloride (DAC) is a drug used to treat hepatitis C virus (HCV) infection. In this study, an ionophore-based nanosphere emulsion was made of tricresyl phosphate (TCP) as the oil phase that is dispersed in water using Pluronic F-127 as an emulsifying agent. The nanospheres, consisting of the oil phase TCP, were doped with sodium tetraphenyl borate (Na-TPB) as a cation-exchanger and dibenzo-18-Crown-6 (DB18C6) as an ionophore (chelating agent) for DAC. The nanosphere emulsion was employed as a titrant in the complexometric titration of DAC (the analyte), and the DAC-selective electrode (ISE) was used as an indicator electrode to detect the endpoint. In the sample solution, DAC2+ ions diffused into the emulsified nanospheres, replaced Na+ from the ion exchanger (Na-TPB), and bonded to the ionophore (DB18C6). The DAC-selective nanospheres were successfully utilized to determine DAC in various samples, including standard solutions, commercial tablets (Daclavirocyrl®), serum, and urine. The method exhibited a linear dynamic range of 81.18 µg/mL to 81.18 pg/mL (10−4 to 10−10 M), achieved high recovery values ranging from 99.4% to 106.5%, and displayed excellent selectivity over similar interfering species (sofosbuvir and ledipasvir). The proposed method offers a new approach to determine the drug species (neutral, anionic, and cationic) without the requirement of water-soluble ligands or pH control.