With the targets of petroleum exploration transferred to the deep and ancient strata, abundant oil and gas resources have been found in Lower Paleozoic and older strata in central and western China. Due to complex evolutionary processes including multiple episodes of hydrocarbon accumulation and ubiquitously accompanied by secondary alterations, significant uncertainties remain concerning the generation time and accumulation processes of these revealed petroleum sources. In this paper, relative pure Re and Os elements existing in the asphaltene fractions of Lower Cambrian solid bitumen collected from the Guangyuan area, western Sichuan Basin, SW China and Middle–Lower Ordovician heavy oils in the Aiding area of the Tahe oilfield in the Tarim Basin, NW China were successfully obtained by sample pretreatments, and Re–Os isotopic analysis was subsequently carried out for the dating of these. The Re–Os isotopic composition indicates a generation time of Guangyuan bitumen to between 572 Ma and 559 Ma, corresponding to the late Sinian period of the Neoproterozoic era. By the means of Re–Os isochron aging, initial 187Os/188Os ratios, and carbon isotopic compositions, the Lower Cambrian bitumen is supposed to originate from source rocks of the Doushantuo Formation in the Sinian strata and subsequently migrated into the reservoirs of the Dengying Formation. This previously reserved petroleum was transformed into its present bitumen state by the destruction of reservoirs caused by tectonic uplift. The Re–Os dating results of Middle–Lower Ordovician heavy oil of Tarim Basin suggest that it was formed between 450 Ma to 436 Ma, corresponding to the Late Ordovician–Early Silurian system, and the generated petroleum likely migrate into the Middle–Lower Ordovician karst reservoirs to form early oil reservoirs. With tectonic uplift, these oil reservoirs were degraded and reformed to the heavy-oil reservoirs of today.