The lipid composition of the tissue is very variable and it also depends on the diet. Lipid composition of different tissues can provide insight into animal nutritional habits, physiological and health status and quality of habitat. The aim of this paper was to determine the lipid composition of the liver, the subcutaneous and perirenal adipose tissue as well as m. gluteus superficialiss, m. semitendinosus, m. vastus lateralis of females and males gray wolf in Croatia and the variation in the composition of fatty substances with regard to sex, body mass and sampling season. The study was conducted on 36 specimens of the gray wolf (14 females and 22 males) harvested during years of legal quota cull. Fatty acid composition was determined by gas chromatography after extraction of fat from tissues, and plasma triacylglycerols, total cholesterol and phospholipids were determined using spectrophotometer. Lipids isolated from liver and muscles of gray wolf were dominated by saturated fatty acids (SFA) while lipids isolated from subcutaneous and perirenal adipose tissue were dominated by dominated by fatty acids with one double bond (MUFA). In m. vastus lateralis higher percentage of C18:0 (p=0,02) and lower percentage of C18:3 (p=0,05) were found in males weighting more than 25 kg mass (+25 kg) compared to males weighting less than 25kg (-25 kg). In m. semintendinosus higher percentage of C16:1c/t (p=0,05) and C18:2 (p=0,04) were found in females weighting more than 25 kg compared to females weighting less than 25 kg. In subcutanous asipose tissue higher percentage of C18:1c (p=0,03) was found in males weighting more than 25 kg compared to males weighting less thta 25 kg. In perirenal adipose tissue higher percentage of C18:2 (p=0,02) was found in females weighting more than 25 kg compared to males weighting more than 25 kg. In liver higher percentage of C20:4n6 (p=0,04) was found in males in the autumn comapred to females in the winter. In m. vastus lateralis higher percentage of MUFA (p=0,04) and C18:0/C18:1 (p=0,03) as well as lower percentage of C18:0 (p=0,01) were found in females in the autumn compared to females in the winter. In m. semintendinosus higher percentage of C18:2 (p=0,03) in females in the winter compared to females in the autumn. Also, males in the autumn had higher percentage of C18:2 (p=0,01) and ratio n6/n3 (p=0,02) compared to females. In m. gluteus superficialis higher percentage of C18:1c/t (p=0,03) and MUFA (p=0,03) were found in females in the autumn compared to females in the winter. In subcutaneous adipise tissue higher percentage of C16:0 (p=0,01) and lower ration n6/n3 were 43 found in males in the autumn compared to males in the winter. Also, higher percenatge of C16:1t (p=0,02) and lower percentage of C20:1 (p=0,02) were found in males in the autumn compared to females. In perirenal adipose tissue higher percentage of C20:4n6 (p=0,04) in females in the winter compared to the females in the autumn. Females in the winter had higher percentage of C20:4n6 (p=0,03) and C22:5 (p=0,03) compared to males. Higher concentration of total cholesterol was found in females in m. vastus lateralis in the autumn compared to males (p=0,03) and higher concentration in m. semitendinosus in females in the winter (p=0,01) compared to males. Based on the results, we can conclude that the presence of fatty acids, for both females and males, differs according to the season, and is dependent on the body mass.