This study evaluated the performance and the quality and fatty acid profile of eggs from laying hens fed diets containing mixed crude glycerin (MCG; 80% vegetable fat + 20% animal fat). A total of 240 39-week-old Hy-Line W36 laying hens were distributed according to a completely randomized experimental design into six treatments consisting of graded MCG dietary inclusion levels (0, 1.5, 3.0, 4.5, 6.0, and 7.5%), with five replicates of eight birds each. Feed intake linearly decreased (p<0.05) with increasing MCG inclusion levels. The percentages of myristic, palmitic, and α-linolenic acids in the eggs linearly decreased as MCG dietary levels increased (p<0.05), while α-linoleic acid, polyunsaturated fatty acids (PUFA) and ω-6/ω-3 ratio linearly increased. Excreta moisture linearly increased with increasing levels of MCG inclusion (p<0.05). MCG may be included in up to 7.5% in layer feeds without impairing performance or egg quality, but levels up to 5.54% reduce SFA egg content. However, the inclusion of MCG in layer diets increases ω-6/ω-3 ratio in the eggs.