This study presents the development and validation of a high-performance liquid chromatography (HPLC) method for the determination of contaminants in drug substances. Ensuring the safety and efficacy of pharmaceutical products is paramount, and the presence of contaminants can significantly impact these factors. The developed HPLC method aims to provide a reliable analytical procedure for detecting and quantifying such contaminants. The method is validated according to International Council for Harmonisation (ICH) guidelines, covering key parameters including specificity, linearity, accuracy, precision, and robustness. The specificity of the method was demonstrated by its ability to distinguish contaminants without interference from other components in the drug substances. Linearity was confirmed through calibration curves with correlation coefficients (R) exceeding 0.999, indicating consistent responses across a range of concentrations (0.1–10 g/mL). Accuracy was assessed via recovery studies, yielding recovery rates between 98% and 102%, thus ensuring the method’s reliability in quantification. Precision, evaluated through repeatability and intermediate precision tests, showed relative standard deviations (RSD) below 2%, affirming the method's reproducibility. Robustness was tested by making deliberate minor changes to method parameters, with results indicating the method's resilience to such variations. This validated HPLC method was then applied to quantify contaminants in commercially available drug substances, demonstrating its practical applicability in real-world scenarios. The study's findings underscore the method's potential for routine quality control in pharmaceutical manufacturing, contributing to enhanced drug safety and efficacy.