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In the present study, sucrose-capped gold nanoparticles (Suc-AuNPs) were synthesized and used as a nanoprobe for the daclatasvir (DAC) detection using its localized surface plasmon resonance (LSPR) property. The determination mechanism was experimentally shown using UV–Vis, dynamic light scattering, and transmission electron microscope (TEM) measurements. The DAC detection is accompanied by the color change from pink to blue following a decrease in the LSPR band of Suc-AuNPs after adding various concentrations of DAC. Under the optimum condition, a good limit of detection (LOD) of 0.008 μg.mL
−1
with a linear concentration response range of 0.01–1.0 µg.mL
−1
was determined at 523 nm for DAC in exhaled breath condensate (EBC) sample. Moreover, the linearity range of 0.1–2.0 µg.mL
−1
and LOD of 0.044 µg.mL
−1
was obtained for DAC in the plasma sample. The employment of Suc-AuNPs for the determination of DAC in EBC samples exhibited high stability, simplicity, and repeatability which can be considered a routine method in clinical laboratories.