The effects of sulfanilic acid, a major azo food dye metabolite, were studied in normal developing rat pups and pups treated with 6-hydroxydopamine (60HDA). Chronic daily intraperitoneal injection of sulfanilic acid during the first postnatal month elicited hyperactivity and impaired shock escape performance in vehicle pups. No differences were noted in 60HDA treated rat pups receiving sulfanilic acid. These findings, which are similar to the results of our study of chronic administration of a food dye mix, suggest that sulfanilic acid may be one of the causative agents in food dye-induced behavioral changes in developing rats. While our work suggests a significant effect of azo food dyes on the developing rat central nervous system, species differences in parameters such as absorption, metabolism, and blood-brain barrier properties do not permit any extrapolation of these observations to proposed effects in children.