We present a study of vanadium (V) isotope compositions for 17 crude oils spanning a wide range of concentrations and formation ages. Bulk organic geochemical and biomarker compositions are investigated for 11 co-genetic crude oils from the Barbados oil field. About 2‰ V isotope fractionation was observed and they are primarily correlated with V/Ni ratios and most likely reflect the depositional environment of the petroleum source rocks. Factors such as the lithology of the source rocks, Eh and pH of the depositional environment, and possibly the V isotope composition of seawater could all play a role in the V isotope composition. The Eh and pH conditions determine the speciation and coordination of V ions in fluids, whereas the lithology of the source rock defines the competing phases for available V ions in solution. The V isotopes are significantly modified by maturation and biodegradation. The V isotope fractionation during biodegradation has most probably resulted from the microbial activity-induced changes in the species and coordination geometries of V ions in fluids. The progressive decrease of δ⁵¹V with the increase of maturation might suggest a preferential loss of ⁵¹V during de-metalation and/or a preferential incorporation of ⁵⁰V in the newly formed V-organometallic compounds.