This study was carried out to determine optimum conditions of five different factors (beta-cyclodextrin concentration, mixing temperature, mixing time, centrifugal force, and centrifugation time) in reduction of cholesterol in 3.6% fat, homogenized milk by application of beta-cyclodextrin. beta-Cyclodextrin at 0.5 to 1.5% provided 92.2 to 95.3% removal of cholesterol when mixed at 10 degrees C for 10 min. Among other factors, mixing time (5 to 20 min) did not significantly affect cholesterol removal. Removal was enhanced with increasing centrifugal forces up to 166x g (95.9%) but decreased thereafter. Various centrifugation times (5 to 20 min) did not have significant effects. Based on these results, we suggest that the optimum conditions for the process are addition of 1.5% beta-cyclodextrin, mixing temperature of 10 degrees C, 10-min mixing time, and centrifugation at 166x g for 10 min.