A novel simple method using liquid chromatography/time-of-flight mass spectrometry (LC/TOF-MS) and LC/UV was established for the quantification of saponins in an extract from green tea leaves. The amount of saponins in a fraction with high in vitro antihypercholesterolemic activity, Fr2-3, was determined to be 72%. An in vivo experiment showed that the addition of 0.5% Fr2-3 to a high-cholesterol diet suppressed the increase in serum cholesterol levels in rats. Fr2-3 induced a decrease in the liver cholesterol and triglyceride levels and an increase in the fecal excretion of cholesterol. These results indicate tea-leaf saponins to be the active components in Fr2-3 and that these saponins exhibited antihypercholesterolemic activity by inhibiting cholesterol absorption in the intestines.