Breast milk contains an amylase that may contribute to carbohydrate digestion in infants. The aim of our study was to determine whether mothers differ in their breast-milk amylase activity and whether the activity is maintained during prolonged lactation. This was investigated by measuring breast-milk amylase activity by hydrolysis of maltotetraose in 63 English mothers of parity 1-5 and 107 Gambian mothers of parity 1-12 who were at various stages of lactation (0.5-27 mo). Amylase was present in all samples and showed a great range of activity from 0.08 to 3.53 IU/mL. Amylase activities did not vary during a feed nor over 24 h, and each mother had a characteristic level of activity in her milk. Amylase activity was higher in the 1st trimester of lactation, and decreased by 35% (p < 0.001) to a plateau at 6-27 mo. Gambian mothers of very high parity (parity 11-12) had 54% of the activity of primiparous mothers (p < 0.001), after adjustment for stage of lactation. Using community data on milk volume, the estimated breast-milk amylase intake by breast-fed children was on the order of 800-1000 IU/24 h in the 1st trimester and 400 IU/24 h in the 2nd year of lactation. Individual measurements emphasized the great differences in this intake among children of the same age. Our study showed that breast milk is an important source of amylase both in developed and developing countries, but there are large variations in intake among children.