Gasoline station attendants spend a great deal of their time in the direct exposure to noxious substances such as benzene and byproducts of gasoline combustion. Such occupational exposure increases the risk of oxidative stress. This study aimed to evaluate hematological and biochemical alterations among petrol station workers. Forty gas station attendants and 39 non-attendants were recruited as exposed and control subjects, respectively. Plasma samples were evaluated for hemoglobin, hematocrit, and red blood cell count via the Sysmex KX-21 analyzer. Then, oxidized hemoglobin, methemoglobin, and hemichrome were measured spectrophotometrically. Moreover, serum antioxidant capacity and protein oxidation were evaluated. The means ± SD of hemoglobin (16.76 ± 0.14 g/dl vs 15.25 ± 0.14 g/dl), hematocrit (49.11 ± 0.36% vs 45.37 ± 0.31%), RBC count (5.85 ± 0.06 mil/μl vs 5.33 ± 0.06 mil/μl), Met-HB (1.07 ± 0.07 g/dl vs 0.39 ± 0.04 g/dl), and hemichrome (0.80 ± 0.07 g/dl vs 0.37 ± 0.02 g/dl) in the exposed group were significantly greater than the control group (P < 0.001). The results of the independent-sample t test illustrated that the FRAP test value in the exposed group (0.23 ± 0.01 mM) was significantly lower than the control group (0.34 ± 0.01 mM), while the value of the plasma protein carbonyl test in the exposed group (7.47 ± 0.33 mmol/mg protein) was meaningfully greater than the control group (5.81 ± 0.19 mmol/mg protein) (P < 0.001). In conclusion, gas station attendants suffer from higher levels of oxidative stress, and they need to take antioxidants in order to minimize the effects of oxidative stress.