The National Children’s Study is a proposed longitudinal cohort study to evaluate the relationships between children’s health and the environment. Enrollment is estimated to begin in September 2005, and 100,000 children will be followed from preconception or early pregnancy until adulthood. Among multiple health outcomes, the study is proposing to investigate whether pre- and/or postnatal exposures to nonpersistent pesticides increase the risk of poor performance on neurobehavioral and cognitive exams during infancy and early childhood. Characterization of exposures will be challenging. Nonpersistent pesticides include many chemicals with biologic half-lives on the order of hours or days. Exposures can occur through multiple pathways (e.g., food and residential or agriculture pesticide use) and by multiple routes (inhalation, ingestion, dermal). Effects may depend on the developmental stage when exposure occurs. Sequential sampling is likely to be required and may involve a combination of environmental and biologic monitoring as well as collection of questionnaire data. In this article we review measurements that can be used to characterize exposures. These include biologic markers, personal and indoor air sampling techniques, collection of dust, surface and dermal wipe samples, and dietary assessment tools. Criteria for sample selection will necessitate evaluation of the time frame of exposure captured by the measurement in relationship to critical windows of susceptibility, the cost and validity of the measurements, participant burden, and variability in exposure routes across populations and at different age periods.