To monitor employees' work safety and exposure against air contaminants, Trans, trans-muconic acid, Hippuric acid, Methyl hippuric acid, Mandelic acid and Phenylglyoxylic acid can be used as reliable biomarkers of exposure to benzene, toluene, ethylbenzene, and xylene (BTEX) compounds. This study aims to determine the level of urinary metabolites of BTEX compounds using biological monitoring in the employees of a wastewater treatment plant (WWTP) in the south of Iran. The study was performed on 56 employees of the WWTP of one of the southern cities of Iran in 2020. Urine samples (n total = 112) consisting of 60 samples of employees working in the operation section (exposed group) and 52 samples of employees working in the administrative section (control group) in the WWTP were collected before and at the end of their shift. The mean concentration of urinary metabolites of BTEX of both groups ranged from 546.43 (μg/g cr) for trans, trans-muconic acid to 0.006 (μg/g cr) for methyl hippuric acid, which indicates that most of the evaluated metabolites showed a higher concentration than their occupational threshold limit value urine (p < 0.05). Regression analysis results showed a significant correlation (p < 0.05) between age and utilization of flame heaters with changes in the measured BTEX metabolites in the urine. The results of this study illustrate that WWTPs should be considered as one of the workplaces with potential sources of BTEX exposure for employees. Future investigations are recommended to perform itemized appraisals of BTEX intake sources, particularly in employees of the operational sections of WWTP.