The fine structures and rheological behaviours of aqueous flour dispersions and of β-glucan, (1→3,1→4)-β- d-glucan isolates obtained from 18 registered varieties of normal covered barley seeds and four registered oat varieties, grown in the same location in Greece, were investigated. The β-glucan content of the barleys and oats varied between 2.5–5.4 and 2.1–3.9%, respectively (dry matter basis). Heat treatment of the barley and oat flour dispersions with 80% (v/v) ethanol, to inactivate endogenous β-glucanases, had a stabilizing effect on the viscosity profile of the flour slurries. The relationship between total β-glucan content and aqueous slurry viscosity (at 247 s −1) of the heat-treated barley flours was weak ( r 2=0.45, p<0.05, n=18). β-Glucans were isolated by water extraction at temperatures slightly below the gelatinization temperature of starch, enzymatic removal of starch and partial removal of contaminating proteins by adjustment to pH 4.0–4.5, and subsequent precipitation of the water-soluble β-glucans with 80% (v/v) ethanol. The cellulosic oligomers released by the action of a (1→3,1→4)-β- d-glucan hydrolase showed cellotriosyl and cellotetraosyl units, accounted for 91.1–92.1% for barley and between 92.4 and 94.1% for the oat preparations; the respective molar ratios of tri- to tetra-saccharides (DP3/DP4) ranged between 2.73–3.05 (barley) and 2.16–2.42 (oat). Steady shear measurements confirmed the random coil type behaviour of freshly prepared β-glucan solutions (5 and 7%, w/v). The rate at which shear thinning began was dependent on both concentration and molecular size of the polysaccharide. Most of the β-glucan dispersions followed the Cox–Merz rule, except Mucio, a variety with high M w β-glucan (2.39×10 5). Viscoelastic characterization, at 8% (w/v), of three barley β-glucan aqueous dispersions differing in molecular size, indicated that the low molecular weight sample exhibited shorter gelation time and higher gelation rate ( I E=[dlog G′/d t] max) than its higher molecular weight counterparts. Small deformation oscillatory measurements on gels of all barley β-glucan isolates (10% (w/v), 7 d storage, 25 °C) revealed a strong negative relationship ( r 2=0.88, p<0.01) between G′ (1 Hz, strain 0.1%) and apparent M w of the polysaccharide.