It was reported that dietary fructose imposes a number of effects on lipid metabolism including hypertriglyceridemia. The daily intake of fructose in humans is mainly due to sucrose. It was reported that the consumption is still increasing, making a background for health implications. The mechanism of metabolic disorders is poorly understood, but a lot of studies indicate that the liver lipid homeostasis deregulation is essential for a fructose effect on metabolism. The aim of the study is to estimate if fructose affects the profile of fatty acids in in vitro and in vivo models. In this study in vitro and in vivo experiments were conducted to assess the effect of dietary fructose on the fatty acid profile in the cell culture or in the liver of rats. The results showed that in the fructose experimental groups, both in the cell and liver homogenates, the content of the saturated fatty acids were significantly higher than in control groups. According to the obtained data fructose in the medium and in the diet affects saturation of fatty acids in the cell cultures and in the livers of rats. The findings obtained in the experiments support the thesis that fructose influences the homeosta- sis of lipid metabolism in the liver and may give an opportunity to discuss the limitation of the content of this kind of sugar in food.