We compare the chromatographic signatures of petroleum biomarkers in Deepwater Horizon (DH) source oil, three other reference crude oils, DH emulsified mousse that arrived on Alabama’s shoreline in June 2010, and seven tar balls collected from Alabama beaches from 2011 to 2012. Characteristic hopane and sterane fingerprints show that all the tar ball samples originated from DH oil. In addition, the diagnostic ratios of various hopanes indicate an excellent match. Quantitation data for C30ab-hopane concentration levels show that most of the weathering observed in DH-related tar balls found on Alabama’s beaches is likely the result of natural evaporation and dissolution that occurred during transport across the Gulf of Mexico prior to beach deposition. Based on the physical and biomarker characterization data presented in this study we conclude that virtually all fragile, sticky, brownish tar balls currently found on Alabama shoreline originated from the DH oil spill. Dataset is associated with the publication Mulabagal, V., Yin, F., John, G., Haysworth, J., & Clement, T., (2013), Chemical fingerprinting of petroleum biomarkers in Deepwater Horizon oil spill samples collected from Alabama shoreline.