This work is an analysis of crude oil samples from the two most recent producing wells in the Machete area (Orinoco Tar Belt, Venezuela). The aim is to determine the type of environment in which the precursor organic materials were deposited. In spite of the fact that the degree of maturity and the biodegradation of the two crude oils are similar, their characterization leads us to conclude that they have very distinctive and different origins. The comparative study was performed by separating their main fractions, pyrolysis of asphaltenes, and further identification of several indicators (aromatic, polar and sulfur compounds, linear aliphatic hydrocarbons, and cadalene). Moreover, several biomarkers (terpanes, hopanoids and steranes) present in the saturated fractions obtained from the two crude oils were analyzed by using gas chromatography combined with mass spectrometry (GC-MS). The statistical study of geochemical indicator values and relative proportions of lipid and lipid-like biomarkers present in products from asphaltene pyrolysis shows a high probability of there being two distinct crude oil families from the Machete area. © 2007 Elsevier B.V. All rights reserved.