Hypocholesterolemic effects of green juice from young barley leaves were investigated on hypercholesterolemia in rats fed on a high cholesterol diet (HCD). The n-hexane extract from a water insoluble fraction of green juice showed hypocholesterolemic activity. At least two substances responsible for the hypocholesterolemic effects were isolated from the n-hexane extract and purified by silica gel chromatography. One of them was β-sitosterol, and the other n-hexacosyl alcohol which was a saturated higher alcohol having a molecular weight of 382. In the rats fed on HCD added with n-hexacosyl alcohol at a concentration of 1%, the plasma cholesterol level was hardly lowered on day 3 but markedly on day 9. In a similar experiment, β-sitosterol markedly lowered the plasma cholesterol level of rats on both day 3 and day 9. Through these experiments, we found that barley leaves contained hypocholesterolemic substances, and that two compounds such as β-sitosterol and n-hexacosyl alcohol were responsible for the hypocholesterolemic activity.