Added PubMed Id Diagnosis of drug-induced liver injury (DILI) and its distinction from other liver diseases are significant challenges in drug development and clinical practice. In this study we identify, confirm, and replicate the biomarker performance characteristics of candidate proteins in a cohort of patients with DILI at onset (DO; n = 133) and follow-up (DF; n = 120), acute non-DILI at onset (NDO; n = 63) and follow-up (NDF; n = 42), and healthy volunteers (HV; n = 104). Area under the receiver operating characteristic curve (AUC) for cytoplasmic aconitate hydratase; argininosuccinate synthase; carbamoylphosphate synthase, fumarylacetoacetase; fructose-1,6-bisphosphatase 1 (FBP1) across cohorts achieves near complete separation (range: 0.94 - 0.99) of DO and HV. In addition, we show that FBP1 alone or in combination with glutathione S-transferase A1 and leukocyte cell derived chemotaxin 2 could potentially assist in clinicla diagnosis by distinguishing NDO from DO (AUC range: 0.65 – 0.78), but further technical and clinical validation of these candidate biomarkers is needed.