Pulses, which are a major component of the Mediterranean diet, are an important source of macronutrients such as proteins, carbohydrates, and dietary fibre [1]. The Food and Agriculture Organization of the United Nations (FAO) declared 2016 the International Year of Pulses, highlighting the importance of these foods not only for their nutritional and healthy aspects, but also for their low environmental impact. Among the various bioactive compounds of legumes, saponins appear to be able to reduce the blood cholesterol levels. In our laboratories have been developed sensitive and specific analytical methodologies for evaluating the level of soyasaponins I and g in lentils by using HPLC-MS/MS equipments. Saponins can be considered components of dietary fibre which are neither digested nor absorbed in the human small intestine [2]; these components together with other carbohydrates present in lentils could possess an important prebiotic action. In this work we have evaluated the hypocholesterolemic and prebiotic activity of a lentil extract by monitoring the plasmatic cholesterol level in an animal model and the concentration of bile acids extrected by faeces, both with the prebiotic effect by using an in vitro model.