This study was carried out to determine optimum conditions (beta-cyclodextrin concentration, mixing time, and holding time) for cholesterol removal from pasteurized nonhomogenized milk at 4 degrees C on a commercial scale by adding beta-cyclodextrin in a specially designed bulk mixer tank. The beta-cyclodextrin (0.4, 0.6, 0.8, and 1.0%) removed from 65.42 to 95.31% of cholesterol at 4 degrees C in 20 min. Treatment of milk with 0.8 and 1.0% (wt/vol) beta-cyclodextrin was no better than treatment with 0.6% beta-cyclodextrin. Maximum cholesterol removal was seen with 6 h of treatment. The beta-cyclodextrin cholesterol complex was precipitated from milk during 20 min without stirring at 4 degrees C and removed by centrifugation. After separating the milk, approximately 0.35% of residual beta-cyclodextrin remained in the skim fraction and 0.1% in the cream from milk treated with 0.6% beta-cyclodextrin. The rest of the beta-cyclodextrin was complexed with the cholesterol and eliminated via the discharger of the separator. Individual fatty acid and triglyceride compositions did not differ between control milk and milk treated with 0.6% beta-cyclodextrin.