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Ora-pro-nóbis (OPN) (Pereskia aculeata Miller) is a non-conventional food plant (PANC) with high nutritional value, rich in proteins, fibers, and minerals, but still scarcely explored by the food industry. This study aimed to evaluate the physicochemical properties of an OPN leaf flour and its application in the development of cookie-type biscuits with different concentrations of this flour. To produce the flour, OPN leaves were dehydrated, ground into flour, and analyzed for proximate composition, instrumental color, water activity, and water and oil absorption and solubility indexes. Four cookie formulations were prepared: one control (without OPN flour) and three with partial replacement of wheat flour by OPN flour at levels of 10%, 20%, and 40% w/w. The cookies were evaluated for proximate composition, texture, color, water activity, and sensory acceptance. The OPN flour showed low water activity (0.1845±0.01), indicating good preservation potential. The water absorption index (6.29±0.36 g/g), oil absorption index (1.91±0.15 g/g), and water solubility index (7.47±0.19 g/g) are important parameters for bakery products. The lipid fraction contributes to flavor and product acceptance, while water absorption enhances yield and dough moisture, improving final quality. In the cookies, increasing the concentration of OPN flour led to higher fiber, ash, and protein contents, highlighting the enhancement of the nutritional value. Regarding the color, the cookies became darker and greener as OPN flour content increased, reflected in the decrease of the L* parameter and in the a* and b* values. In terms of texture, the 40% w/w OPN flour formulation had greater hardness (208.38±59.37 N), likely due to the higher fiber content, which reduces softness and increases crunchiness. In the sensory analysis, the 10% w/w and 20% w/w OPN flour formulations showed good acceptance, with average scores above 6 (“slightly liked”) for appearance, aroma, flavor, texture, and overall acceptance. It can be concluded that OPN flour is a promising alternative to nutritionally enhance baked products such as cookies. However, further studies are needed to optimize the sensory aspects of these products, reinforcing the role of OPN in promoting healthier and more functional eating habits.
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