The metabolism of β-sitosterol was compared to that of cholesterol in 12 patients. Sterol balance methods were supplemented by radiosterol studies, with the following results. (a) Plasma concentrations of β-sitosterol ranged from 0.30 to 1.02 mg/100 ml plasma in patients on intakes of β-sitosterol typical of the American diet. Plasma levels were raised little when intakes were increased greatly, and on fixed intakes they were constant from week to week. On diets devoid of plant sterols, the plasma and feces rapidly became free of β-sitosterol. (b) The percentage of esterified β-sitosterol in the plasma was the same as for cholesterol. However, the rate of esterification of β-sitosterol was slower than that for cholesterol. (c) Specific activity-time curves after simultaneous pulse labeling with β-sitosterol-³H and cholesterol-¹⁴C conformed to two-pool models. The two exponential half-lives of β-sitosterol were much shorter than for cholesterol, and pool sizes were much smaller. Values of turnover for β-sitosterol obtained by the sterol balance method agreed closely with those derived by use of the two-pool model. There was no endogenous synthesis of β-sitosterol in the patients studied; hence, daily turnover of β-sitosterol equaled its daily absorption. Absorption of β-sitosterol was 5% (or less) of daily intake, while cholesterol absorption ranged from 45 to 54% of intake. (d) About 20% of the absorbed β-sitosterol was converted to cholic and chenodeoxycholic acids. The remainder was excreted in bile as free sterol; this excretion was more rapid than that of cholesterol. (e) The employment of β-sitosterol as an internal standard to correct for losses of cholesterol in sterol balance studies is further validated by the results presented here.