Premature infants are susceptible to disease related to deficient dietary calcium intake. Studies in adults suggest carbohydrates can enhance calcium absorption. However, little is known about how carbohydrates affect calcium absorption in premature infants due to a lack of direct in vivo studies. We adapted the triple lumen perfusion method for use in premature infants to compare calcium absorption 36 mmol/L (1.44 g/L) in the absence and presence of either 70 g/L lactose or glucose polymers. 44Ca was added to determine endogenous calcium losses. Fourteen infants were studied (gestational age: 31 +/- 0.4 wk; study weight: 1590 +/- 105 g; mean +/- SEM). Calcium absorption from the glucose polymer solution was greater than that from the control and lactose solutions (0.17 +/- 0.05 mumol.min-1.cm-1 versus 0.04 +/- 0.04 and 0.008 +/- 0.045 mumol.min-1.cm-1, respectively). Calcium absorption correlated positively with water and carbohydrate absorption. The rate of carbohydrate absorption was greater from the glucose polymers than from the lactose solution (0.40 +/- 0.10 mg.min-1.cm-1 versus 0.22 +/- 0.06, respectively). Based upon 44Ca absorption, endogenous calcium loss appeared to account for less than 1% of total calcium flux. We conclude that glucose polymers, but not lactose, enhance calcium absorption in the premature infant, a fact that may be useful in formula design.