Male Wistar rats were fed on a conventional diet containing normal corn starch or 6% enzyme-resistant starch originating from either raw or retrograded high-amylose corn starch. Furthermore, the diets were either cholesterol-free or contained 1% cholesterol and 0.1% cholic acid. The main objective of this study was to investigate whether the addition of enzyme-resistant starch to a rat conventional diet had any effect on cholesterol metabolism. Therefore, plasma and liver cholesterol concentrations, plasma HDL:LDL cholesterol ratios and neutral steroid and bile acid excretion were determined. No significant effect of enzyme-resistant starch feeding on plasma and liver cholesterol concentrations was found. However, consumption of raw or retrograded high-amylose corn starch resulted in a decrease in esterified and total liver cholesterol concentrations of 24 and 22%, respectively. This was accompanied by a reduction in plasma esterified and total cholesterol levels of 4% and a tendency to higher daily faecal coprostanol and total bile acid excretion.