Herbicides are plant protection products used to control weeds. They are used in agriculture and were available for use by the general public until January 1, 2019. They are widely present in our everyday environment. In France, following the Pélagie mother-child cohort—which provided the first exposure data in Brittany—and the perinatal component of the national biomonitoring program, the Esteban cross-sectional study (Health Study on the Environment, Biomonitoring, Physical Activity, and Nutrition) made it possible to measure, for the first time, herbicide exposure levels in the French mainland population (adults and children). These analyses were conducted using a subsample of 891 adults and 498 children included in the study between April 2014 and March 2016. Only two herbicides were actually quantified in Esteban participants: 2,4-D and glyphosate. AMPA was the only herbicide metabolite quantified, and it was detected in over 70% of adults and children. The levels of AMPA found in children aged 6 to 17 were slightly higher than those measured in adults. The levels of 2,4-D exposure found in the Esteban study were lower than those found in North American studies. One explanation for this finding could be higher herbicide use in the United States and Canada. These two countries are the second and fifth largest users of herbicides in the world, respectively, while France ranks ninth globally. Regarding the results for glyphosate and AMPA, the urinary concentrations observed in Esteban appeared similar to those observed in European studies. AMPA exposure increased with the use of a standalone boiler as the primary heating source, meat consumption, and wine consumption. As for children, their exposure was higher in the spring than in the fall or winter. Finally, the levels of herbicides measured in urine cannot be compared to toxicological reference values and are therefore not predictive of an associated health risk. Furthermore, based on current knowledge, there is little data on the health effects of herbicides at low exposure levels and no health threshold value established by the agencies responsible for setting such thresholds.