Toads of Rhinella jimi species (Stevaux, 2002), belong to Bufonidae family and are endemic of the Brazilian northeastern, commonly founded in rainy periods. In general, amphibians of this family present in their venoms different metabolites which have a diversity of pharmacological activities. The isolation and identification of these compounds are of great importance and techniques such as high-performance liquid chromatography coupled to mass spectrometry are widely used for the discovery of novel and known compounds in these venoms. For R. jimi venom, were obtained the ethyl acetate and methanolic extracts and thirty compounds tentative identified by HPLC-MS and MS/MSn, which marinobufagin (2) and marinobufotoxin (19) were the majorities for each extract, respectively. In addition, other bufadienolides mainly presented in the ethyl acetate extract, such others bufotoxins, alkaloids and arginine diacid derivatives were tentative identified in the methanol extract. In cytotoxic assay by MTT, the extracts and compound 2 demonstrated IC50 values below or near to positive control doxorubicin, demonstrating excellent cytotoxic potential and for antimalarial assay, the samples were tested against Plamodium falciparum, demonstrating a parasitemia reduction (%) in range of 81-90%, being all samples active. This is a general report of the chemical composition of the venom from R. jimi toad venom and it’s respective cytotoxic and antimalarial activities, promoting the enrichment knowledge of family and species.