Herbicide 2,4-Dichlorophenoxyacetic acid (2,4-D) and its analogues are widely used in agriculture. Although the occurrence of 2,4-D in urine has been widely reported in North America, it has scarcely been investigated in China, especially in young children. In addition, both in vivo and in vitro studies have shown that high-level 2,4-D exposure is associated with oxidative stress, but their association in a general sensitive population has rarely been evaluated. In this study, 2,4-D and its analogues were measured in 417 urine samples collected from 139 children aged 0–7 during the non-peak season of pesticide application in Wuhan, central China, and Shenzhen, south China. Each of them provided three samples in three consecutive days. An oxidative stress biomarker, 8-hydroxy-2-deoxyguanosine (8-OHdG), was also measured. The geometric mean (GM) of unconjugated urinary 2,4-D concentration was 0.10 μg/L (corrected by urinary specific gravity, SG-corrected). After β-glucuronidase hydrolysis, the GM of SG-corrected urinary 2,4-D was 0.15 μg/L, and the detection frequency was 100%. Moderate inter-day reproducibility was found within individuals, with an intraclass correlation coefficient of 0.68 for SG-corrected urinary deconjugated 2,4-D. The GM of estimated daily intake of 2,4-D was 6.05 ng/kg-bw/day. A significant positive correlation was found between urinary 2,4-D and 8-OHdG, whereas no association was found after SG-correction. This is the first study to characterize the occurrence of urinary 2,4-D, its inter-day reliability, and its association with urinary 8-OHdG in young children from China.