IntroductionDoxorubicin‐induced cardiotoxicity (DIC) is a serious obstacle to oncologic treatment. Mountain cultivated ginseng (MCG) exhibits stronger pharmacological effects than cultivated ginseng (CG) mainly due to the differences in ginsenosides. However, the material basis and the underlying mechanism of the protective effects of total saponins of MCG (TSMCG) against DIC are unclear.ObjectivesWe aimed to elucidate the material basis and the pharmacodynamic effects of TSMCG on DIC as well as the underlying mechanisms.MethodsTo comprehensively analyze the effective substances, the chemical components of TSMCG and their prototypes or metabolites in vivo were characterized through UHPLC/Q‐TOF‐MS. Then, an absorbed component–target–disease network was established to explore the mechanisms underlying the protective effects of TSMCG against DIC. H9c2 cells were employed for pharmacodynamic assays. The mechanism was verified by Western blot and molecular docking simulations.ResultsA total of 56 main ginsenosides were identified in TSMCG, including 27 ginsenosides of PPD type, 15 ginsenosides of PPT type, two ginsenosides of OA types, and 12 ginsenosides of other types. Moreover, 55 ginsenoside prototypes or metabolites in vivo were tentatively characterized. Ginsenoside Ra1, a differential compound between MCG and CG, could be metabolized by oxidation and deglycosylation. Network pharmacology showed that AKT1, p53, and STAT3 are core targets of 62 intersecting genes. Molecular docking results indicated that most of the ginsenosides have favorable affinity with these core targets. After doxorubicin exposure, TSMCG could increase cell viability and inhibit apoptosis in a dose‐dependent manner.ConclusionOur work reveals a novel comprehensive strategy to study the material basis of the protective effects of TSMCG against DIC and the underlying mechanisms through integrating in vivo substance identification, metabolic profiling, network pharmacology, pharmacodynamic evaluation, and mechanism verification.