The present study was carried out to evaluate the effect of different lines and generations of adult males and females of Japanese quail (Coturnix japonica) on total lipid, fatty acid (FA) composition and cholesterol content of breast muscle, as well as the identification of quantitative trait loci (QTLs) controlling the above mentioned meat quality traits. Forty-four quails (generation F 0 ), 22 Pharaoh (F-33) meat-type males and 22 Standard (S-22) laying-type females, were crossed to produce the F 1 hybrids generation. F 2 generation was created by mating one F 1 male with one F 1 female, full siblings. The birds, randomly chosen from F 0 (22 males and 22 females), F 1 (22 males and 22 females) and F 2 (84 males and 152 females), were raised to 20 weeks of age in collective cages. After slaughter the carcasses were dissected and the breast muscle was removed for the chemical analysis. Genomic DNA was extracted from the whole blood and 30 microsatellite markers located on two quail chromosomes were genotyped. S-22 quails exhibited higher amounts of total saturated fatty acids (SFA) and polyunsaturated fatty acids (PUFA), while F-33 quails had a greater content of total monounsaturated fatty acids (MUFA). F-33 quails supplied meat with lower atherogenic and thrombogenic indexes. A partial effect of sex on the FA composition of quail meat was found in the F 1 generation; while in the F 2 generation a sex-effect was more evident. F 2 females exhibited a higher PUFA content and PUFA/SFA ratio, and a lower muscle cholesterol content compared to F 2 males. In conclusion, total lipid, FA composition and cholesterol content were affected by first-and second-generation crosses. For the cholesterol content a QTL showing additive effect has been detected on CJA02 at 85 cM; no QTLs were found for total SFA, MUFA and PUFA. To current knowledge, this is the first study of a QTL associated with muscle cholesterol in quails.