<p>Drug-induced hepatotoxicity remains a critical challenge in pharmacological safety evaluation; however, the lack of analytical tools to distinguish cell-type-specific hepatotoxic responses hinders mechanistic understanding of toxicity pathways. Herein, we provide proof-of-principle for using an immunocompetent liver-on-a-chip platform that integrates six distinct cell models with a targeted cell depletion strategy to rapidly assess pharmaceutical candidates’ hepatotoxic profiles. The platform recapitulated immune cell migration dynamics and stress-responsive behaviors under chemokine induction, while integrated fluidic characterization ensured physiologically relevant microenvironmental control. We applied the targeted depletion strategy to resolve cell population-specific contributions to hepatotoxic outcomes using four mechanistically diverse compounds: acetaminophen, ethinyl estradiol, sulfamethoxazole, and abacavir. Our findings reveal coordinated participation of multiple hepatic cell populations in toxicological processes and identify dominant hepatotoxicity-inducing factors. The immune-dependent toxicity resolution was further validated using a known immune-mediated drug-induced liver injury drug, allopurinol. Overall, this study establishes a targeted cell depletion strategy integrated within an immunocompetent liver-on-a-chip platform, enabling rapid identification of hepatotoxic determinants, and represents a significant advancement in predictive toxicology systems, while providing strategic insights for subsequent pathway analysis.</p>