This study describes the progressive changes in exocrine pancreatic enzyme specific activities from rats provided with a diet containing either 62% starch (S) or fructose (F) with 6.0 ppm copper (+Cu) or 0.6 ppm (-Cu) from 21 to 61 days of age. Copper-deficient rats had significantly reduced pancreatic weight and DNA, RNA, protein and copper content and superoxide dismutase (SOD) activity after consuming diet for 39 days. These reductions were far more severe in F-Cu than in S-Cu rats. Copper deficiency significantly reduced pancreatic amylase and lipase specific activities, but the effect was more severe in the F-Cu dietary group. Chymotrypsinogen and trypsinogen specific activities were reduced only in the F-Cu dietary group and the CHO x Cu interaction was significant. High dietary fructose and low dietary copper in combination interact to produce severe reductions in pancreatic amylase, lipase, chymotrypsinogen, and trypsinogen specific activities which could not be explained by either copper deficiency or by an increase in dietary fructose alone.