Asphaltite is a crude oil transformed through geological processes into a solid material with stable chemical and physical properties, which makes it an economical commodity. It occurs in significant quantities in the Upper Cretaceous to Miocene strata of the Middle Magdalena Valley Basin (MMVB) in Colombia, where a network of asphaltite dikes is locally mined. This study deals with the origin and distribution of these asphaltites and places it in the context of the regional petroleum system. Geological mapping of the asphaltite dikes allowed them to be placed in the context of the area's structural deformation history. Different analytical techniques have been employed to determine the chemical and physical conditions, including open-system pyrolysis, gas chromatography–mass spectrometry (GC-MS), and inductively coupled plasma mass spectrometry (ICP-MS). Rock-Eval results gave a Total Organic Carbon content of 77.9% to 99.7%, a Hydrogen Index of 579 to 671, an Oxygen Index from 10 to 18, and Tmax of 438 to 448, suggesting the dikes are composed of Type II kerogen and are in the early to peak mature oil window. The asphaltites are composed of asphaltenes (79%), resins/NSO (16%), polycyclic aromatic hydrocarbons (3%), and alkanes (2%), with notably high concentrations of vanadium (7000 ppm), nickel (2500 ppm), chromium (33 ppm), and lithium (9 ppm). The organic geochemical analyses include: Pr/Ph ratio suggest dominantly reducing conditions of deposition; the proportion of the C27, C28, and C29 steranes and the H30/H29 ratio both indicate a mixture of organic matter sources, while the Ts/Tm ratio supports the thermal maturity of the analyzed samples in the early mid-oil window. The molecular composition of the asphaltite dikes, adjacent oil seeps, and samples of the La Luna Formation are highly similar, supporting the hypothesis that the La Luna Formation represents the primary source rock. The emplacement of the asphaltite dikes is linked to the Andean tectonic event and preferentially occurs in the eastern foothills of the MMVB, associated with fractures and faults along the Bucaramanga–Santa Marta strike-slip system. This study provides the first detailed geochemical characterization and genesis of Colombian asphaltites, demonstrating their link to the petroleum system of the MMVB.