Background and objective: Benzene is one of the main pollutants in air and one of the most extensive chemical compound used in both natural and industrial processes. Benzene exposure leads to the most dangerous adverse health effects, particularly blood cancer. The aim of this study was to evaluate the gas station workers’ exposure to benzene by measuring benzene in breathing air and urinary trans, trans-muconic acid. Materials and Methods: This cross-sectional study conducted in summer 2014 investigated 40 gas station workers and 40 occupationally non-exposed persons. Spot urine samples were obtained prior to and at the end of the work shift from each subject. The urinary levels of trans, trans-muconic acid was determined by liquid chromatography (HPLC) with diode array detector. At the end of the work shift, the benzene collected on sorbing cartridges was desorbed using carbon disulfide and was analyzed using gas chromatography and mass spectrometry detection. Results: The mean value for exposure to benzene in breathing zone of gas station workers was 5.90 ±1.93 ppm, which was significantly greater than the occupationally non exposed group (1.15±0.744 ppm). The mean urinary concentrations of trans, trans-muconic acid differed significantly between samples of gas station workers (64.75±19.47 µg/L) and occupationally non-exposed persons (47.10±13.67 µg/L). Conclusion: A good correlation (r = 0.581) between the mean values of benzene in breathing zone and the urinary concentration trans, trans-muconic acid was observed. Gas station workers were found to be probably the most exposed groups in this study. Inhalation is presumably the main route of exposure in gas station workers.