Preterm and term transitional milks of human subjects and mature milks of human subjects, non-human primates and non-primates were analysed for free amino acids (AA) using precolumn phenylisothiocyanate derivatization and liquid chromatography. Differences in free AA between three types of human milk were small. Milks of pinnipeds (seals and sea lions) contained the highest levels of total free AA (8634-20,862 mumol/l), while the milks of cows and sheep had the lowest levels of total free AA (1061-1357 mumol/l). The milks of human subjects, chimpanzees (Pan troglodytes), gorillas (Gorilla gorilla), elephants (Elephas maximus), horses and pigs had intermediate levels of total free AA (3069-7381 mumol/l). Glutamic acid was the most abundant free AA in milks of human subjects (1339-2157 mumol/l), non-human primates (423-2528 mumol/l), elephants (1332 mumol/l), horses (1119 mumol/l), and cows (349 mumol/l). Taurine was the most abundant free AA in milks of pinnipeds (5776-13,643 mumol/l), pigs (1238 mumol/l), goats (1150 mumol/l) and sheep (341 mumol/l). Taurine was the second most abundant free AA in milks of human subjects and non-human primates, while histidine was the second most abundant free AA in milks of pinnipeds. Milks of each species had a distinctive free AA pattern which may reflect the relative importance of the free AA during early postnatal development.