Oats (Avena satica L.) are an important source of dietary fiber mainly composed of β-glucans, which is reported to be effective in lowering of cholesterol and decreasing the risk of heart attack. Subcritical water extraction (SWE) is a new and promising method for the extraction of β-glucan due to its polarity-selectivity, efficiency of recovery, time-efficiency, and lower cost. In this study, efficiencies of the SWE were investigated for the extraction of β-glucan from oat flour. The conditions for maximizing the extraction efficiency were determined by varying temperature (110-190°C), pH of solvent (pH 3.0-10.6), static time (5-20 min) and particle size of oat flour. The amount of β-glucan was determined with a ‘Mixed-linkage β-glucan’ assay kit (Megazyme International Ltd.), according to McCleary and Codd (1991). The overall results showed that the concentration of β-glucan was highest under the following conditions: extraction temperature of 200°C, pH value of 4.0, static time of 10 min, and particle size of 425-850 μm. The extraction yield under the optimum condition was 6.98±1.17 g/100 g oat flour, which was over two-fold higher than that obtained using hot water extraction method (60°C, 3 h). Therefore, SWE is a feasible alternative for extracting soluble dietary fiber (β-glucan) from oat flour.