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Abstract: A total of 45 alkylbenzenes were detected and identified in crude oils with different depositional environments and thermal maturities from the Tarim Basin, Beibuwan Basin, and Songliao Basin using comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC?×?GC-TOFMS). By analyzing the distribution characteristics of C0–C5 alkylbenzenes, it is found that the content of some alkylbenzenes varies greatly in crude oils. Based on the distribution characteristics of 1,2,4,5-tetramethylbenzene (TeMB) and 1,2,3,4-TeMB, the ratio of 1,2,4,5-TeMB to 1,2,3,4-TeMB is proposed to indicate the organic matter origin and depositional environment of ancient sediments. Oil samples originated mainly from lower hydrobiont, algae, bacteria and source rocks deposited under reducing/anoxic conditions have low 1,2,4,5-/1,2,3,4-TeMB values (less than 0.6), while oil samples originated mainly from terrestrial higher plants and source rocks deposited under oxic/sub-oxic conditions have higher 1,2,4,5-/1,2,3,4-TeMB values (greater than 1.0). The significant difference of 1,2,4,5-/1,2,3,4-TeMB values is controlled by 1,2,4,5-TeMB content. 1,2,4,5-TeMB content in oils derived from source rocks deposited in oxidized sedimentary environment (greater than 1.0?mg/g whole oil) is higher than that in oils from source rocks deposited in reduced sedimentary environment (less than 1.0?mg/g whole oil). 1,2,4,5-/1,2,3,4-TeMB ratio might not or slightly be affected by evaporative fractionation, biodegradation and thermal maturity. 1,2,4,5-/1,2,3,4-TeMB ratio and 1,2,4,5-TeMB content can be used as supplementary parameter for the identification of sedimentary environment and organic matter input. It should be noted that compared to the identification of organic matter sources, the 1,2,4,5-/1,2,3,4-TeMB parameter is more effective in identifying sedimentary environments.