Seven crude oil samples from Los Manueles field, Maracaibo Basin, Venezuela, were analyzed to evaluate oil mixtures associated with different oil charges into the reservoirs. Analyses of the bulk physicochemical parameters suggest variations in the API gravity, the concentration of saturated and aromatic hydrocarbons, NSO compounds + asphaltenes, sulfur, vanadium, nickel, and the biomarkers distribution. The oils were divided in two groups: G-I with high saturated hydrocarbons and low NSO compounds + asphaltenes, sulfur, vanadium and nickel, a bimodal distribution of n-alkanes, high Pr/Ph and oleanane index, low steranes index, low abundance of tricyclic terpanes and C35S/C34S < 0.54 and G-II with low saturated hydrocarbons and high NSO compounds+ asphaltenes, sulfur, vanadium and nickel, an unimodal distribution of n-alkanes with low Pr/Ph and oleanane index, high steranes index, abundance of tricyclic terpanes and C35S/C34S > 0.70. G-I has a higher contribution of terrigenous organic matter compared to G-II, characterized by a higher contribution of marine organic matter. Source rock lithology biomarker indicators suggest marine shale as the source rock for oils of terrigenous origin. The results indicate that oils were generated by two independent source rocks, La Luna Formation and a secondary source rock with terrigenous organic matter, probably represented by the Capacho Formation.