Eggs are highly nutritious however the challenge of cholesterol has been a major concern for human consumption. The effects of graded levels of vitamin C on yolk, serum and cholesterol levels of layers were tested. Eighty (80) Isa brown layers of 28 weeks old were used for this study. The hens were grouped into 4 treatments namely T1(control), T2, T3 and T4 with 0, 100, 150 and 200 mg vitamin C respectively in commercial layer mash. Both feed and water were given ad libitum. At 32 and 36 weeks of age, 6 eggs from each group were selected and broken to separate the yolk from the albumen for cholesterol analysis, likewise, 6 hens from each group were bled and the sera analysed for cholesterol level. Data obtained were analysed statistically using one –way ANOVA. Vitamin C reduced yolk and serum cholesterol levels (P<0.05). Serum cholesterol levels in mm/mol were T1 = 3.86, T2 = 3.62, T3 = 2.92 and T4 = 1.12 while yolk cholesterol for T1, T2, T3 and T4 were 105.25, 61.47, 48.66 and 37.75 mm/mol respectively. Vitamin C supplementation in layers’ feed was beneficial to the hens since it also helped in producing low cholesterol eggs for human consumption with very levels.