The drug toxicity is a long-term concern in contemporary medical research. Serious drug toxicity may cause acute liver failure or even patient death. Currently, the pathogenesis of drug-induced liver injury (DILI) is not entirely clear, and there is still no specific treatment for patients with DILI. Accordingly, improving the diagnosis and treatment level of DILI is a major challenge facing the basic and clinical research in relevant fields. To address these, an ·OH-activated circular dichroism (CD) and photoacoustic (PA) dual-mode nanoprobe was here designed and synthesized. The probe was prepared using chiral d/l-cysteine and polymolybdate as raw materials to form a nanocomplex with chiral properties (Ox-POM@d/l-Cys) based on the interaction between metal ions and sulfhydryl groups in Ox-POM@d/l-Cys. In vitro and in vivo experimental results have shown that the as-proposed dual-mode nanoprobe can be used not only for CD spectral detection of Fenton’s reagent but also for PA imaging monitoring of ·OH. More importantly, inspired by the NIR PA properties, the Ox-POM@d/l-Cys probe was used for the first time to detect ·OH in DILI mice. This will provide very useful information for the diagnosis and treatment of DILI disease.