Abstract: Objective: This study was conducted to determine the effect of chili leaf on laying hen performance, egg quality and egg yolk cholesterol levels. Materials and Methods: The completely randomized design involved evaluation of a control (no chili leaf powder) diet and experimental diets supplemented with chili leaf powder (CLP) at 1, 2 or 3%. One hundred and twenty laying hens (Charoen Pokphand Brown) at 61 weeks of age were divided into 4 treatments, each with 10 replicates (3 birds per replicate). Each group was randomly allocated to one of the treatments for 5 weeks. All diets were isonitrogenous and isocaloric. Results: The results of the effect of supplementation with CLP at 0, 1, 2 and 3% showed that the body weight gain, egg yield, egg weight and cholesterol were not significantly different among the treatments (p>0.05). However, supplementation with CLP at 0 and 1% resulted in a higher feed intake and feed conversion ratio than those obtain with CLP supplementation at 3% (p<0.05). There were no significant differences among treatments in egg quality, breaking strength, shell thickness, yolk color, yolk percentage, shell percentage, albumen percentage, Haugh unit, lightness (L*) of yolk color, redness of yolk color (a*) and yellowness (b*) of yolk color (p>0.05). Conclusion: Dietary supplementation with 3% CLP can be used as alternative feed additive for laying hens despite an adverse effect on feed intake and feed conversion ratio, as feeding 3% CLP had no adverse effects on egg quality.