Purpose <p>Acute leukemia is the most commonly diagnosed malignancy in children. In addition to genetic factors, exposure to chemicals also increases its risk. Considering the increasing incidence of pediatric acute leukemia and limited experience on the underlying role of environmental pollutants, this study aims to assess the relationship between exposure to benzene and organochlorine compounds and pediatric acute leukemia.</p> Methods <p>Participants of this cross-sectional study were children aged under 15 years diagnosed as new cases of acute leukemia and healthy children. Urine samples were collected for measurement of benzene and organochlorine pesticides metabolites by using liquid-liquid microextraction techniques.</p> Results <p>This study comprised 496 participants including 195 (39.31%) children in the leukemia group and 301 (60.69%) in the healthy group. The majority of participants were male (63.1%). The leukemia and healthy groups differed significantly in their urinary levels of α- and β-Hexachlorocyclohexane (<i>P</i> = 0.001), endosulfan (<i>P</i> = 0.002), aldrin (<i>P</i> = 0.002), and heptachlor (<i>P</i> = 0.001) after the adjustment of urine creatinine. Furthermore, adjusted logistic regression analysis revealed that elevated levels of β-BHC was significantly associated with an increased risk of acute leukemia.</p> Conclusion <p>This study provides findings that warrant further investigation into the significant association between exposure to benzene and organochlorine and acute leukemia in children. Further research with a larger sample size and a prospective design is necessary to obtain more conclusive results. Environmental policies for reducing the exposure to chemicals should be considered as a health priority.</p>