A simple, new, green, and sensitive approach was established and validated for assay of the recently approved antineoplastic medication; darolutamide (DAR) in its authentic form, pharmaceutical formulation, and biological fluids fluorimetrically. This experiment relied on the native fluorescence of the cited drug and detects the ideal solvent utilized throughout the approach. The proposed approach was validated regarding linearity, accuracy, and precision. The calibration graph showed linearity over the range of 0.1–2.0 μg mL−1. The limit of detection and quantitation (LOD and LOQ) were 0.032 μg mL−1 and 0.09 μg mL−1, respectively. Because of the approach's high sensitivity, it was decided to spike the mentioned drug in plasma and urine samples. At last, checking for content uniformity was performed regarding the United States Pharmacopoeia (USP) by adjusting the proposed approach.
