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Comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC–TOFMS) was used for geochemical characterization of 18 oil samples from Campos Basin, Brazil. Conventional analyses were also performed on these oils (API gravity and GC-FID) and in the maltenic fraction (saturated, aromatic, and polar analysis) after asphaltene precipitation, aiming the oil screening for a rapid assessment of general characteristics. The results from principal component analysis with the biomarker parameters of source, maturity, and biodegradation obtained by GC×GC–TOFMS separated the oils into two groups, mainly explained by gammacerane content. The higher sensitivity and resolution of GC×GC–TOFMS allowed the identification of unusual compounds in oils from this basin, such as methylhopanes (whose calculated ratios allowed the oils in this work to be classified as having a marine, lacustrine, or “mixed” source, the same interpretation obtained by statistical analysis), moretanes (with results that reinforce the hypothesis of same thermal evolution for the studied samples), and short-chain steranes (C21 and C22), detected in very low concentrations in all of the samples. This study is the first to show the presence of these compounds in Campos Basin crude oils.