The relation between indices of neonatal iron status and individual differences in neonatal temperament were investigated in a sample of 148 low-income Peruvian women and their newborn infants. Using cord blood, at birth we obtained measures of neonatal ferritin, serum iron, and hemoglobin. While neonates were still in the hospital, their behavior during a structured anthropometry examination was videotaped and subsequently coded on four temperament dimensions: activity level, negative emotionality, alertness, and soothability. The same dimensions were coded using a videotape obtained during a subsequent visit to the neonates' homes. Results indicated that lower levels of neonatal hemoglobin and serum iron were related to higher levels of negative emotionality and to lower levels of alertness and soothability. A similar pattern was found for ferritin, but only for females. For the most part, relations between neonatal iron measures and neonatal temperament were linear, operating across the full range of iron values. Our pattern of iron–temperament results could not be attributed to variation in family demographics, low birth weight, gestational age, maternal dietary intake, or markers of neonatal illness and maternal diabetes. Our findings are consistent with prior research with older infants relating iron deficiency to temperament. These results support the importance of increased research on the early functional–behavioral consequences of individual differences in iron status as well as on the mechanisms that underlie such consequences. © 2005 Wiley Periodicals, Inc. Dev Psychobiol 46: 141–153, 2005.