It has been suggested that phytosterol and phytostanol esters possess similar cholesterol-lowering properties, however, whether mechanisms responsible are identical has not been addressed. To address this question, cholesterol plasma levels, absorption, biosynthesis, and turnover were measured in 15 hypercholesterolemic subjects consuming prepared diets each over 21 d using a crossover design. Diets contained either i ) margarine (M), ii ) margarine with phytosterol esters (MSE) (1.84 g/d), or iii ) margarine with phytostanol esters (MSA) (1.84 g/d). Cholesterol absorption was measured using the ratio of [ 13 C]cholesterol oral :D 7 -cholesterol IV ; biosynthesis using D incorporation from D 2 O and turnover by D 7 -cholesterol IV decay rates. Plasma total cholesterol level at d 21/22 was lower ( P Ͻ 0.05) for MSE (13.4%) but not MSA (10.2%) versus M (6.0%) diets. Plasma low density lipoprotein-cholesterol (LDL-C) mean reductions at d 21/22 were larger ( P Ͻ 0.05) for MSE (12.9%) and MSA (7.9%) compared with M (3.9%). Plasma TG and high density lipoprotein-cholesterol (HDL-C) levels did not differ across diets. Cholesterol absorption was reduced ( P Ͻ 0.05) 36.2 and 25.9% at d 21 for MSE and MSA versus M, while cholesterol biosynthesis was reciprocally increased ( P Ͻ 0.05) 53.3 and 37.8% for MSE and MSA versus M, respectively. Cholesterol turnover was not influenced by diet. These data indicate that plant sterol and stanol esters differentially lower circulating total and LDL cholesterol levels by suppression of cholesterol absorption in hypercholesterolemic subjects. -