Background: Drug-induced liver injury (DILI) is a significant clinical challenge, often leading to acute liver failure and the withdrawal of pharmaceutical agents from the market. Metabolomic profiling has emerged as a promising tool to identify biomarkers for early detection of hepatotoxicity. This study aims to investigate the biochemical alterations associated with DILI in commonly prescribed pharmaceuticals using metabolomic analysis.Materials and Methods: A total of 60 adult patients, categorized into drug-exposed (n=40) and control (n=20) groups, were enrolled. Blood and urine samples were collected at baseline, 4 weeks, and 12 weeks following drug administration. High-performance liquid chromatography-mass spectrometry (HPLC-MS) and nuclear magnetic resonance (NMR) spectroscopy were employed for metabolomic profiling. Key biochemical markers, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, and metabolic signatures of hepatotoxicity, were analyzed using multivariate statistical techniques.Results: Significant alterations in metabolite profiles were observed in the drug-exposed group compared to controls. Elevated levels of ALT (72 ± 15 U/L), AST (89 ± 18 U/L), and total bilirubin (2.4 ± 0.6 mg/dL) were detected in patients exhibiting hepatotoxicity. Metabolomic analysis revealed an upregulation of lipid peroxidation markers and a reduction in glutathione levels (1.8 ± 0.4 µmol/L). Principal component analysis (PCA) demonstrated distinct clustering between hepatotoxic and non-hepatotoxic patients, suggesting a predictive role of metabolic perturbations in DILI.Conclusion: Metabolomic profiling provides a biochemical approach to predicting DILI, offering potential biomarkers for early detection and risk assessment. The findings highlight the need for metabolomics-based screening in drug safety evaluation to minimize hepatotoxic risks in clinical practice.