Aerobic biodegradation of crude oil and its pathways were investigated via in vitro culture and GC-MS analysis in water flooding wells of Dagang oil field. The in vitro aerobic culture lasted 90 days when 99.0% of n-alkanes and 43.03-99.9% of PAHs were degraded and the biomarkers and their ratios were changed. The spectra of components in the residual oil showed the similar biodegradation between aerobic process of 90 days and degradation in reservoir which may last for some millions years, and the potential of serious aerobic biodegradation of petroleum in reservoir. 24 Metabolites compounds were separated and identified from aerobic culture, including fatty acid, naphthenic acid, aromatic carboxylic acid, unsaturated acid, alcohols, ketones and aldehydes. The pathways of alkanes and aromatics were proposed, which suggests that oxidation of hydrocarbon to organic acid is an important process in the aerobic biodegradation of petroleum. (C) 2013 Elsevier Ltd. All rights reserved.