Shatt Al-Arab River is one of the important rivers for water supply, industry and irrigation in Basrah City, south of Iraq. It's also supply drinking water to users in the region. Despite the significance of this river, there is a lack of previous studies dealing with petroleum pollution especially for geochemical studies of organic matter in the river sediments using molecular tracer methods. Therefore, the identification and quantification of petroleum hydrocarbons in this river at the elemental and molecular levels will be of the utmost interest. Sediments were analyzed in nine Shatt Al-Arab River stations. Texture was silty clay or silty sand. TOC and TN ranged from 0.39-0.90 and 0.02-0.16%. OM varied from 17.43-34.36 µg g −1 dw, representing 0.27-0.87% of TOC. AHs ranged from 3.19-10.27 µg g −1 dw and constituted 16.07-29.88% of OM. Total n-alkanes concentrations ranged from 0.08-42.58 µg g −1 dw with carbon numbers from C11-C34 were identified. N-alkanes content was relatively moderate compared to sites contaminated with oil worldwide. Dominance of even carbon number n-alkanes (C11-C20) with CPI and LMW/HMW values close to unity, indicated to oilrelated n-alkanes. This was verified by the values of U/R (1.27-2.45), pri/phy (0.82-1.00), C17/pri (0.70-1.94) and C18/phy (0.95-1.56) ratios and the presence of UCM. Biogenic contributions were detected within the n-alkanes distribution of C17-C31 and CPI and LMW/HMW values of <1 that mainly related to terrestrial plants, phytoplankton, algae and bacteria sources, confirmed by TAR (0.79-2.15) and C31/C19 ratio (0.28-1.62) values. However, the contribution from biogenic hydrocarbons is overshadowed by petroleum origin n-alkanes. This was obvious by the presence of UCM. PCA showed concordant results with n-alkanes ratios indicating mixed n-alkanes sources in sediments. Hopanes and steranes in sediments displayed similar signatures that were characteristic of mature organic matter contribution from oil contaminations.