The purpose of this study was to investigate the relationship among frequency of sucrose exposure, mutans streptococci levels and dental caries. Adult volunteers took part in this crossover study performed in 4 phases of 28 days each. The volunteers wore intra-oral palatal appliances containing blocks of human dental enamel and dripped 20% sucrose solution onto the dental blocks from 0 to 8 times/day. After each phase, the colony forming units (CFU) were determined in dental plaque and enamel dental caries was evaluated using cross-sectional hardness. Sucrose frequency had no statistically significant effect on mutans streptococci levels. In the enamel cross-sectional hardness tests, significant differences (p < 0.05) in relation to area of mineral loss were observed only when sucrose exposure was 8 times/day. Similar results were obtained when cross-sectional hardness was assessed at each distance from enamel surface.