The objective of this study was to determine the distribution of (1-3)(1-4)-β-d-glucans in the kernels of three high-glucan lines of naked barley (STH 4561, STH 4671 and STH 4676), a low-glucan line (STH 4572), and two low-glucan comparative cultivars (naked – Rastik, and hulled – Stratus). The content of (1-3)(1-4)-β-d-glucans assayed with the enzymatic method varied within the range from 4.04 to 5.71% dry matter basis. Microspectrophotometric analysis revealed significant differentiation in the structure of the kernels and in the distribution of (1-3)(1-4)-β-d-glucans; it also indicated a necessity of applying such processing methods as are appropriate to the distribution of functional components. The distribution of functional components in the high-glucan lines of naked barley makes them ideal for utilization both in the milling technology and in the production of grits and flakes.