This work focuses on the use of high-performance liquid chromatography (HPLC) for the quantitative analysis and validation of propylthiouracil tablets through reverse phase chromatography (HPLC-RP). The analysis was optimised using a C18 column with a mobile phase that includes acetonitrile and buffer in a ratio of 20:80 (v/v). The buffer is made from monobasic potassium phosphate with a pH of 4.6 ± 0.05. The detection of propylthiouracil was performed at ? = 272 nm. Samples were extracted using methanol and water. The newly developed method exhibited good linearity between 24.916 and 74.748 µg/mL, with an R2 > 0.999. Precision expressed in terms of % relative standard deviation (RSD) was within the acceptable range, whereas accuracy in terms of % recovery varied between 98-102%. Therefore, the proposed and validated HPLC-RP method is reliable for the quantitative analysis of propylthiouracil in pharmaceutical formulations, ensuring accurate dosage determination and quality control. The technique shows promising potential for pharmacokinetic studies and routine quality assurance in the pharmaceutical industry. © 2025 Elsevier B.V., All rights reserved.