This study aimed to examine the possibility of the production of bread using whole barley flour (WBF); two levels of substitution of 50% and 100% compared with wheat flour (WWF) only were administered.  The effect of the baking process on the content of dietary fiber, particularly β-glucan, was investigated. Chemical composition alongside with the content and solubility of b-glucan in both raw materials and bread produced were determined. The results obtained showed that the barley flour gave the highest content of ash, crude fiber and fat compared to wheat flour. The highest content of total and soluble beta-glucan were recorded in whole barley bread followed by 50% barley substituted  treatment. Moreover, the 100%WBF bread scored the highest content of fiber, protein and ash as well as fat content compared with wheat bread only, which was the highest in carbohydrate content only. Content of β-glucan showed higher values ​​for the bread produced from whole barley flour followed by treatment 50% of barley flour and then bread traditional wheat flour, which scored the least content of β-glucan. The process of fermentation and baking could significantly reduce the total content of β-glucan in spite of increased solubility of beta-glucan. Taking into account the functional dose of β-glucan, it could be assumed that, presented content of β-glucan in barley bread, meet the recommended daily needs to achieve the health benefits attributed to barley β-glucan.