Bioavailability of alimentary components is an important indicator at the estimation of the influence of the nutrition on the human health. Taking into account the difficulty in accessing to intestinal contents in vivo, some artificial models enabling research on the intestinal digestion and absorption in vitro were elaborated. This paper also presents structure and physiology of gastrointestinal tract in vivo. Many detailed data on composition and concentration of active substances in alimentary canal, kinetics of pH changes of intestinal fluids as well as the rate of passage and digestion of food in gastrointestinal tract are discussed. Basic models of gastrointestinal tract in vitro used for the investigation of the intestinal digestion and the absorption are also described. The majority of GI models consist of two – or three-stage systems including: stomach – small intestine, mouth – stomach – small intestine or stomach – small intestine – large intestine. The optimal weight (volume) ratio between different components of intestinal fluids: food/saliva/gastric juice/bile/pancreatic juice is 1.5/1/2/1/2. To study the transport of food compound across intestinal epithelium, two types of in vitro models are used: dialysis or ultrafiltration membranes and cell cultures of intestinal epithelium in vitro. The most commonly used cell line to study intestinal absorption is human intestine cell line Caco-2. Its morphology and physiology is very similar to the human small intestine enterocyte cells in vivo. The Caco-2 model was applied to study absorption of protein, carbohydrates, lipids, vitamins, metal ions, antioxidants, mycotoxins and other food ingredients.