The use of fruit industrial waste in the processing new foods represents an important new step for the food industry. This study aimed to develop cookie recipes using different amounts of guava peel flour (GPF) levels (30%, 50%, and 70%) to evaluate the proximate composition, and the phenolic compound, lycopene, and β-carotene levels in the cookies and flour and to evaluate the cookie sensory acceptance. The results demonstrated low moisture, lipid and carbohydrate contents in the flour and cookies. GPF was considered rich in fiber, ash, polyphenols, and β-carotene. The sensory analysis showed satisfactory acceptance of the cookies containing 30% GPF regarding the aroma, flavor, and texture attributes. The cookies containing 50% and 70% GPF received satisfactory acceptance regarding to aroma only. In conclusion, GPF can be used to partially replace wheat flour in the preparation of cookies to improve the nutritional quality without affecting the product sensory quality.