Background: Consumption of 3 g oat b-glucan/d is considered sufficient to lower serum LDL cholesterol, but some studies have shown no effect. LDL cholesterol lowering by oat b-glucan may depend on viscosity, which is controlled by the molecular weight (MW) and amount of oat b-glucan solubilized in the intestine (C). Objectives: Our 2 primary objectives were to determine whether consumption of 3 g high-MW oat b-glucan/d would reduce LDL cholesterol and whether LDL cholesterol lowering was related to the log(MW · C) of oat b-glucan. Design: In a double-blind, parallel-design, multicenter clinical trial, subjects with LDL cholesterol 3.0 and 5.0 mmol/L (n = 786 screened, n = 400 ineligible, n = 19 refused, n = 367 enrolled, and n = 345 completed) were randomly assigned to receive cereal containing wheat fiber (n = 87) or 3 g high-MW (2,210,000 g/mol, n = 86), 4 g medium-MW (850,000 g/mol, n = 67), 3 g medium-MW (530,000 g/mol, n = 64), or 4 g low-MW (210,000 g/mol, n = 63) oat b-glucan/d (divided doses, twice daily) for 4 wk. Results: LDL cholesterol was significantly less with 3 g high-MW, 4 g medium-MW, and 3 g medium-MW oat b-glucan cereals than with the wheat-fiber cereal by 0.21 (5.5%; 95% CI: 20.11, 20.30; P = 0.002), 0.26 (6.5%; 95% CI: 20.14, 20.37; P = 0.0007), and 0.19 (4.7%; 95% CI: 20.08, 20.30; P = 0.01) mmol/L, respectively. However, the effect of 4 g low-MW oat b-glucan/d (0.10 mmol/L) was not significant (2.3%; 95% CI: 0.02, 20.20). By analysis of covariance, log(MW · C) was a significant determinant of LDL cholesterol (P = 0.003). Treatment effects were not significantly influenced by age, sex, study center, or baseline LDL cholesterol. Conclusions: The physicochemical properties of oat b-glucan should be considered when assessing the cholesterol-lowering ability of oat-containing products; an extruded breakfast cereal containing 3 g oat b-glucan/d with a high-MW (2,210,000 g/mol) or a medium-MW (530,000 g/mol) lowered LDL cholesterol similarly by '0.2 mmol/L (5%), but efficacy was reduced by 50% when MW was reduced to 210,000 g/mol. This trial was registered at www.clinicaltrials.gov as NCT00981981.Am J Clin Nutr 2010;92:723-32.