The purpose of this study was to compare the levels of acetylcholinesterase (AChE) among employees based on job positions and the wearing of personal protective equipment (PPE) in fuel station areas. The sample group consisted of 200 people sorted into various groups, including (i) Inside fuel dispenser area (I-FDA) group consisting of 100 employees for refueling, cashier, and loading fuel into storage tanks, and (ii) Outside fuel dispenser area (O-FDA) group, consisting of 100 employees working in convenience stores, food stores, coffee shops, and offices, as well as general staff and car washers. Data were collected using interview questionnaires and blood samples were analyzed for AChE activity. The results showed that the I-FDA group had a significantly lower mean of serum AChE (7.38±1.73 U/ml) than the O-FDA group (7.85±1.49 U/ml) (p<0.05). The I-FDA group had a 2.43 times higher risk of abnormal serum AChE than the O-FDA group (OR=2.43, 95%CI=1.05-5.60). When considering the risk factors for job positions, it was found that refueling and cashier positions had lower AChE activity levels than those who did not (p<0.05). In the part, employees for food sales in a closed building and those wearing PPE masks had significantly higher AChE activity levels than those who did not or who wore them (p<0.05). Based on the aforementioned, employers at fuel stations should strictly determine the appropriate measures for wearing a PPE mask. Further, employees should be provided accommodation away from oil supply or enclosed buildings during breaks to reduce the effects on the neurotransmitter.