Objectives Approximately 13% of all births occur prior to 37 weeks gestation in the U.S. Some established risk factors exist for preterm birth, but the etiology remains largely unknown. Recent studies have suggested an association with environmental exposures. We examined the relationship between preterm birth and exposure to a commonly used herbicide, atrazine, in drinking water. Methods We reviewed Kentucky birth certificate data for 2004–2006 to collect duration of pregnancy and other individual-level covariates. We assessed existing data sources for atrazine levels in public drinking water for the years 2000–2008, classifying maternal county of residence into three atrazine exposure groups. We used logistic regression to analyze the relationship between atrazine exposure and preterm birth, controlling for maternal age, race/ethnicity, education, smoking, and prenatal care. Results An increase in the odds of preterm birth was found for women residing in the counties included in the highest atrazine exposure group compared with women residing in counties in the lowest exposure group, while controlling for covariates. Analyses using the three exposure assessment approaches produced odds ratios ranging from 1.20 (95% confidence interval [CI] 1.14, 1.27) to 1.26 (95% CI 1.19, 1.32), for the highest compared with the lowest exposure group. Conclusions Suboptimal characterization of environmental exposure and variables of interest limited the analytical options of this study. Still, our findings suggest a positive association between atrazine and preterm birth, and illustrate the need for an improved assessment of environmental exposures to accurately address this important public health issue.