Lipids are essential and integral components of membrane and perform important functions in the biological objects – energy source, biological active substance. The goal of the experiments was to determine the lipid composition of rooster and boar sperm under cryopreservation. Plasmatic membranes were isolated using a three-phase polymeric system by the method improved by authors. The lipids were studied using the micro-analytical method of thin-layer chromatography. The experimental results allowed to isolate and identify in a fresh-obtained and thawed after freezing rooster sperm the following components: phospholipids, 1,3-diglycerides, cholesterol and triglycerides. The lipid spectrum of plasmatic membranes of thawed boar sperms was represented as follows: phospholipids – 18,4+0,16; 1,2-diglycerides – 6,5+0,06; 1,3-diglycerides – 9,1+0,04; cholesterol – 5,1+0,23 and triglycerides – 60,9+0,33 %. The comparative analysis of densitograms of lipid components in plasmatic membranes of rooster and boar sperms confirms the qualitative peculiarities of the lipid structure. Peak of phospholipids, 1,3-diglycerides and cholesterol were detected in rooster sperm, and in boar sperm – the peak of phospholipids, 1,3-diglycerides and triglycerides. The characteristic feature in this case are the absence of 1,2-diglycerides and a considerable decrease of triglycerides amount in rooster sperm after freezing-thawing. According to the latest data, phospholipids are relatively unstable cell structural components. We have identified six phospholipid fractions – sphingo-myelin, phosphatidileholine, phosphatidilserine, phosphatidilethanolamine, cardiolipine and phosphoric acid in plasmatic membranes of native and frozen-thawed rooster spermatozoa. Cryopreservation process leads to practical absence of cardiolipine fraction. The obtained data allow us to assume that since the amount of phosphatidilethanolamine (the most unsaturated phospholipid) increases in a preserved sperm more than twice, the response of rooster sperm to low temperatures leads to the increase of fatty unsaturated acids amount, which can be considered as an adaptation-compensation reaction. In boar cryopreserved sperm the phospholipids are represented by four fractions with the absence of phosphatidilserine and cardiolipin. The comparison of the obtained chromatograms shows qualitative and quantitative peculiarities of phospholipids structure of sperms of studied animals. Thus among phospholipids of rooster and boar sperm membranes there is a large amount of phosphatidilcholine, phosphatidilethanolamine and phosphatidic acid, in boar sperm phosphatidic acid is found even in higher amount. This fact indicates a more substantial damage of boar sperm membranes since its presence suggests the possibility of reparation processes in spermatozoa membranes during cryopreservation. At the same time a minor component of phospholipids, in an equal amount in both species, is the sphingomyelin, which is known as the most saturated phospholipids. The generalization of the obtained research results makes possible the conclusion that the quantitative changes of lipid spectrum of plasmatic membranes in rooster and boar spermatozoa under the influence of low temperature are the most pronounced in phospholipid and cholesterol fractions.