Vitamin B-6 status, assessed by plasma pyridoxal phosphate (PLP) concentrations, and vitamin B-6 concentrations in breast milk were examined in 47 lactating mothers supplemented with different amounts of pyridoxine·HCl (PN·HCl) during pregnancy and the first 6 mo of lactation. PLP concentrations in cord blood and maternal plasma at 2 d postpartum and vitamin B-6 concentration in colostrum were positively correlated with the amount of PN·HCl supplementation prenatally (r = 0.71, p < 0.001; r = 0.74, p < 0.001; and r = 0.78, p < 0.001, respectively). Correlations between the amounts of PN·HCl supplementation postnatally and plasma PLP concentrations increased with the length of supplementation. Plasma PLP concentrations were also correlated with vitamin B-6 concentrations of milk samples, which were obtained on the same day as plasma. PN·HCl supplements between 2.5 and 4.0 mg/d (2.1-3.4 mg PN equivalents) ensured vitamine B-6 adequacy of the mother and maintained relatively saturated concentrations of vitamin B-6 in breast milk.