Thiamin deficiency remains an important public health problem in some populations. The aim of the present investigation was to study thiamin status during the third trimester of pregnancy and its influence on the concentration of this vitamin in transition (days 13–14 of lactation) and mature breast milk (day 40 of lactation) in a group of Spanish women. The pregnancies and lactation periods of fifty-one healthy women 18–35 (mean 26·7 (sd 3·7)) years old were monitored. Vitamin intake during the third trimester was determined by recording the consumption of foods over 5 d and of the quantities provided by dietary supplements. Thiamin status during this stage of pregnancy was determined by measuring the activation coefficient of erythrocyte transketolase (α-ETK). Milk thiamin content was estimated (in 41% of the subjects) by oxidizing thiamin to thiocrome and measuring fluorescence. Subjects with thiamin intakes above that recommended (group H) had more satisfactory serum α-ETK coefficients (1·01 (sd 0·19)) than did those with lower intakes (group L) (1·21 (sd 0·30); P < 0·05). Mature milk thiamin concentrations were significantly higher in group H subjects (0·59 (sd 0·44) μmol/l) than group L subjects (0·25 (sd 0·07) μmol/l). Subjects with α-ETK coefficients > 1·25 in the third trimester had significantly lower mature milk thiamin concentration (0·31 (sd 0·10) μmol/l) than did subjects with more satisfactory α-ETK levels at this time (0·55 (sd 0·42) μmol/l; P < 0·05). The thiamin status of women can be improved since 25·5% of subjects took less than that recommended and 13·7% showed signs of severe deficiency (α-ETK > 1·25). The influence of maternal thiamin intake on α-ETK coefficients and on mature breast milk thiamin concentration is confirmed.