The purposes of the present study were to determine whether caffeine has an effect on zinc absorption and tissue levels of Zn. Under anaesthesia, one side of the intestine of female rats was connected to infusion pumps and an infused solution (first caffeine and later Zn solution) was collected from the other side of the intestine using either 300 mm or the whole small intestine to determine Zn absorption. In a further study, different doses of caffeine solution were injected directly into the femoral vein and a saline or Zn solution was infused into the intestine to measure Zn absorption and tissue Zn concentrations. The results consistently showed that the caffeine solution infused into the intestine did not affect intestinal Zn absorption nor was absorption affected by the direct injection of caffeine into the vein. In contrast, injection of different doses of the caffeine solution significantly decreased Zn tissue levels for the heart only. Calcium concentrations in the heart tissue were also decreased, but not magnesium concentrations. Tissue Zn levels recovered immediately on infusion of a Zn solution into the intestine.