The effects of concentration, temperature, and time on infusion of fluorescein into corn and waxy rice starches and their controlled release pattern were investigated. At low fluorescein concentration (1 μM), temperature significantly affected infusion efficiency. At high fluorescein concentration (50-150 μM), temperature showed little effect; fluorescein concentration significantly affected infusion efficiency. Corn starch showed relatively higher infusion efficiency than waxy rice starch at high concentration. During controlled release, 50% and 81% of infused fluorescein were released from corn and waxy rice starches, respectively, after bacterial α-amylase treatment. However, 61% and 68% of infused fluorescein were released from corn and waxy rice starches, respectively, after pancreatic α-amylase treatment. The dextrose equivalent (DE) value revealed similar patterns, suggesting that degradation of starch by different α-amylases is a major factor affecting release of fluorescein from starch granules. Moreover, granule size of starch greatly affected enzymatic hydrolysis and controlled release in this system. The online version contains supplementary material available at 10.1007/s10068-022-01059-2.