An HPLC method for the determination of β-glucan in barley was developed. The β-glucan was hydrolysed with lichenase [endo-β-(1-3),(1-4)-d-glucan-4-glucanhydrolase from Bacillus subtilis] to oligosaccharides, which were analysed by reversed-phase HPLC using water as the mobile phase at a flow-rate of 0.7 ml/min. The separation of the oligosaccharides was performed in a C 1 8 stainless-steel column (Spherisorb ODS-2) with 5-μm particles in less than 10 min, with a refractive index detection.