Aurantio-obtusin　is　an　anthraquinone　derived　from　Cassia　obtusifolia　(cassiae　semen).　It　is　also　used　as　a　tool　and　a　detection　index　for　the　identification　of　cassiae　semen,　as　stipulated　by　the　Chinese　Pharmacopoeia.　Anthraquinones,　the　main　components　in　cassiae　semen,　have　been　reported　to　show　hepatotoxicity.　This　study　investigates　the　hepatotoxicity　of　aurantio-obtusin　in　male　Sprague-Dawley　rats.　We　randomly　divided　the　animals　into　a　blank　control　group　and　treated　three　test　groups　with　different　doses　of　aurantio-obtusin:　Low　dose　(4　mg/kg),　medium　dose　(40　mg/kg),　and　high　dose　(200　mg/kg).　Each　group　was　treated　with　aurantio-obtusin　for　28　days,　whereas　the　control　group　was　administered　an　equal　volume　of　0.5%　carboxymethyl　cellulose　sodium　salt　(CMC-Na)　aqueous　solution.　Subsequently,　we　conducted　biochemical,　hematological,　and　pathological　investigations　and　determined　the　weight　of　different　organs.　We　used　serum　metabolomics　to　identify　possible　biomarkers　related　to　hepatotoxicity.　The　low-dose　group　showed　no　significant　liver　injury,　whereas　the　medium-　and　high-dose　groups　manifested　obvious　liver　injury.　Compared　with　the　control　group,　the　test　groups　showed　an　increase　in　alanine　transaminase,　aspartate　transaminase,　and　alkaline　phosphatase　levels.　The　liver　organ　coefficient　also　significantly　increased.　Additionally,　we　found　significant　changes　in　the　hematological　indices.　Metabolomics　analysis　showed　that　aurantio-obtusin　induced　28　endogenous　markers　related　to　liver　injury.　Our　data　indicate　that　aurantio-obtusin　induces　hepatotoxicity　in　rat　liver　in　a　dose-dependent　manner　and　is　mediated　by　pathways　involving　bile　acids,　fatty　acids,　amino　acids,　and　energy　metabolism.　In　particular,　changes　in　bile　acid　content　during　treatment　with　therapeutic　agents　containing　aurantio-obtusin　deserve　increased　attention.