The present study was carried out to evaluate the effect of partial replacement of wheat flour with either Jerusalem artichoke powder, barley flour and a mixture of both at different levels. Proximate composition and sensory evaluation were determined. Moreover, organoleptic evaluation proved that the substitution of Jerusalem artichoke powder. The barley flour and a mixture of both was the best as it received high scores by the panelists. The present study also aimed to evaluate the biochemical parameters of diabetic rats fed on some functional food mainly Jerusalem artichoke tubers and barley. Glucose level, total cholesterol, liver function, kidney function, total protein, triglycerides, high density lipoprotein (HDL) and low density lipoprotein (LDL) in diabetic rats serum were also estimated. The results indicated that the bread substituted by Jerusalem artichoke powder, barley flour, and a mixture of both induced a significant decrease in serum glucose levels, triglycerides, total cholesterol and LDL-cholesterol of rats in the hyperglycemic groups, in comparison with rats in the positive control group that were fed on different but not the same prepared diet.