We have studied net uptake of zinc from segments of rat duodenum, jejunum, and ileum using in vivo intestinal perfusion. We have also evaluated the effect of pancreatic and bile secretions on duodenal zinc absorption. Segments of duodenum, jejunum, and ileum and duodenum with bile and pancreatic duct obstruction, 10 cm in length, were studied in six rats each. Percentage absorption of zinc as determined by atomic absorption spectrophotomety was greater from the ileum (60.1%) when compared to duodenum (19.1%) or jejunum (20.2%). Exclusion of bile and pancreatic secretions from the lumen increased zinc absorption in the duodenum (32.0%). Postperfusion mucosal zinc content comprised 29.0% of absorbed zinc from jejunum, 7.4% from ileum, 5.2% from duodenum, and 2.7% from duodenum with bile and pancreatic duct obstructed, indicating rapid transport of zinc across epithelial cells after uptake from the duodenum and ileum. Our data indicate that the ileum has the greatest capacity for zinc absorption.