Abstract Bitumen found in Neoproterozoic carbonates from the southern Amazonian Craton, Brazil, represents a great challenge for its geochemical characterization (origin, thermal maturity and the degree of preservation) within a context of petroleum system. This organic material occurs in the basal Araras Group, considered as a Neoproterozoic cap carbonate, composed of dolostones (Mirassol d’Oeste Formation) overlaid by limestones and shales (Guia Formation). Geochemical analyses in samples of carbonate with bitumen from two open pits (Terconi and Tangara quarries) have shown low to very low total organic carbon content. Analyses of representative samples of Guia and Mirassol d’Oeste formations allowed us to obtain Gas chromatography (GC) traces and diagnostic biomarkers. n -C 14 to n -C 37 alkane distribution patterns in all samples suggests a major contribution of marine algae. Mid-chain monomethyl alkanes (C 14 C 25 ) identified in both sets of samples were also reported in all mid to late Proterozoic oils and source rocks. However, there are significant differences among terpane distribution between the Mirassol d’Oeste and Tangara da Serra regions. The integration of organic geochemistry data and geological information suggests an indigenous origin for studied bitumen, primarily accumulated as hydrocarbon fluids migrated to carbonate rocks with higher porosity and permeability, and afterwards, altered to bitumen or migrabitumen. Although further investigations are required, this work provides a significant contribution to the knowledge about the remnant of this hypothetical Neoproterozoic petroleum system developed in the Southern Amazonian Craton.