Evaluation of protein quality and safety of high-fat cookies made from composite flours of hildegardia barteri, maize, and plantain using in vivo feeding trials Evaluation of protein quality and safety of high-fat cookies
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Abstract
This study evaluated the protein quality and safety of high-fat cookies produced via the creaming method from composite flours of maize, (Hildegardia barteri) and plantain. In a controlled feeding trial with 42 albino rats, diets formulated with the cookies were assessed for key nutritional and safety parameters. The results indicated that the cookies supported significant rat growth, with high protein quality evidenced by a biological value of 60–84%, net protein utilization of 50–78.6%, and true protein digestibility of 83.6–95%. Rats fed Diets 1, 2, and 3 demonstrated superior nutrient utilization and higher activity levels, with Diet 3 performing most favorably. In contrast, Diets 4, 5, and 6 resulted in lower nutrient absorption and reduced activity. All diets were found to be non-toxic, as confirmed by relative organ weights (kidney, liver, heart) and serum biochemical markers (AST, ALT, ALP) that remained within normal physiological limits. These findings demonstrate that composite flour cookies are a safe and nutritious food option, highlighting their potential to enhance food security and contribute to healthier diets.
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References
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Musigwa S., Cozannet P., Asiamah C.A., Wu S.B. Effects of dietary protein levels, net energy levels, and essential amino acid-to-true protein ratios on broiler performance. Animals, 2024, 14(21): 3065. https://doi.org/10.3390/ani14213065
Nychyk O., Barton W., Rudolf A.M., Boscaini S., Walsh A., Bastiaanssen. Protein quality and quantity influence the effect of dietary fat on weight gain and tissue partitioning via host-microbiota changes. Cell Reports, 2021, 35(6): 109093. https://doi.org/10.1016/j.celrep.2021.109093
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Olanrewaju O.O., Fafiolu A.O., Oso A.O. Nutritive value of animal protein sources in rat model: A review. Nigerian Journal of Animal Production, 2024, 51(1): 123-134. https://doi.org/10.51791/njap.vi.7153
Otoo E., Asibuo J.Y., Akromah R. Enhancing protein and vitamin A intake in maize-based porridge using orange-fleshed sweet potato and cowpea composite flour. Legume Science, 2024, 6(2): e231. https://doi.org/10.1002/leg3.231
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Roschke N.N., Hillebrandt K.H., Polenz D., Klein O., Gassner J.M.G.V., Pratschke J., Krenzien F., Sauer IM., Raschzok N., Moor S. Optimizing environmental enrichment for Sprague Dawley rats: Exemplary insights into the liver proteome. PLOS ONE, 2024, 19(4): e0297497. https://doi.org/10.1371/journal.pone.0297497
Salles J., Guillet C., Le Bacquer O., Malnero-Fernandez C., Giraudet C., Patrac V., Berry A., Denis P., Pouyet C., Gueugneau M., Boirie Y., Jacobs H., Walrand S. Pea proteins have anabolic effects comparable to milk proteins on whole-body protein retention and muscle protein metabolism in old rats. Nutrients, 2021, 13(12): 4234. https://doi.org/10.3390/nu13124234
Schipper L., Oosting A., Scheurink A.J.W., van Dijk G., van der Beek E.M.A. customized grain-based diet as an alternative to the AIN-93 diet for mice. PLOS ONE, 2024, 19(3): e0298765. https://doi.org/10.1371/journal.pone.0298765
Sobotka W., Drażbo A. The effect of dietary protein restriction in phase feeding systems on nitrogen metabolism and excretion in pig production. Animals, 2025, 15(11): 1521. https://doi.org/10.3390/ani15111521
Terefe Z.K., Omwamba M., Nduko J. Effect of solid-state fermentation on nutritional and antinutritional properties of maize flour. Food Science & Nutrition, 2021, 9(9): 5221-5231. https://doi.org/10.1002/fsn3.2485
Tesalonika T., Sianipar O., Suryanto E. Advancements in rodent housing systems: Welfare and research implications. Livestock and Animal Research, 2025, 23(1): 45-56. https://doi.org/10.20961/lar.v22i3.84171
Udedeh H.E., Emelike N.J.T., Okorie-Humphrey C., Enyi C.U. Nutritional and sensory properties of cookies from African walnut (Tetracarpidium conophorum), Justicia carnea, and wheat flour. IPS Applied Journal of Nutrition, Food and Metabolism Science, 2024, 2(1): 19-26. https://doi.org/10.54117/iajnfms.v2i1.49
Warrilow A., Mellor D., McKune A.J., Pumpa K. Dietary fat, fibre, satiation, and satiety: A systematic review of acute studies. European Journal of Clinical Nutrition, 2019, 73(3): 333-344. https://doi.org/10.1038/s41430-018-0295
Akinwotu K.O., Taiwo K.A., Ikujenlola A.V. Advancements in composite flours: Applications, nutritional benefits, and processing techniques. Asian Food Science Journal, 2025, 24(6): 24-36. https://doi.org/10.9734/afsj/2025/v24i6792
Akinyede A.I., Ayibiowu E.O., Fakologbon T., Awolu O., Fagbemi T. Nutritional assessment, glycemic indices and anti-diabetic potentials of dough meal generated from optimized blends of matured plantain, soya cake and wheat bran flours. Journal of Future Foods, 2023, 3(4): 374-382. https://doi.org/10.1016/j.jfutfo.2023.03.008
Akinyemi T.Y., Akinyede A.I., Oluwajuyitan T. Extruded breakfast cereal from finger millet flour blends: Nutritional composition, in-vivo protein quality assessment, and biochemical indices of rats fed. NFS Journal, 2022, 29(11): 35-42. https://doi.org/10.1016/j.nfs.2022.10.001
Alahmari L.A. Dietary fiber influence on overall health with an emphasis on CVD, diabetes, obesity, colon cancer, and inflammation. Frontiers in Nutrition, 2024, 11(12): 1510564. https://doi.org/10.3389/fnut.2024.1510564
Amusa H.O., Kehinde R., Ayotebi R.O., Abu O.A. Haematology, serum biochemistry and organ weight changes of Wistar rats fed processed dehulled jack bean (Canavalia ensiformis). Nigerian Journal of Animal Production, 2020, 42(1): 71-78. https://doi.org/10.51791/njap.v42i1.830
Anyadioha J.I., Attaugwu R.N. Physicochemical and sensory evaluation of cookies from germinated maize (Zea mays)–kpaakpa (Hildegardia barteri)–blanched plantain (Musa paradisiaca) composite flour. Toros University Journal of Nutrition and Gastronomy, 2023, 2(2): 137-149. https://doi.org/10.58625/jfng-2283
Anyiam P.N., Phongthai S., Sai-Ut S., Kingwascharapong P., Jung Y.H., Zhang W., Rawdkuen S. Nutritional components and digestibility profiles of some potential plant-based protein sources. Foods, 2025, 14(10): 1769. https://doi.org/10.3390/foods14101769
AOAC International. Official methods of analysis of AOAC INTERNATIONAL (22nd ed.). AOAC International, 2023.
Berghaus D., Haese E., Weishaar R., Sarpong N., Kurz A., Seifert J., Camarinha-Silva A., Bennewitz J., Chillon T., Stefanski V., Rodehutscord M. Nitrogen and lysine utilization efficiencies, protein turnover, and blood urea concentrations in crossbred grower pigs at marginal dietary lysine concentration. Journal of Animal Science, 2023, 101: skad335. https://doi.org/10.1093/jas/skad335
Berrazaga I., Micard V., Gueugneau M., Walrand S. The role of the anabolic properties of plant-versus animal-based protein sources in supporting muscle mass maintenance: A critical review. Nutrients, 2019, 11(8): 1825. https://doi.org/10.3390/nu11081825
Biradar S.D., Kotecha P.M., Godase S.N., Chavan U.D. Studies on nutritional quality of cookies prepared by maida and little millet flour. International Journal of Chemical Studies, 2021, 9(1): 1675-1680. https://doi.org/10.22271/chemi.2021.v9.i1x.11466
Calvez J., Azzout-Marniche D., Tomé D. Protein quality, nutrition and health. Frontiers in Nutrition, 2024, 11(5): 1406618. https://doi.org/10.3389/fnut.2024.1406618
Chung Y., Gulumian M., Pleus R., Yu I. Animal welfare considerations when conducting OECD test guideline inhalation and toxicokinetic studies for nanomaterials. Animals, 2022, 12(23): 3322. https://doi.org/10.3390/ani12233322
de Almeida Moreira L.F., de Lima A.C.S., da Silva B.P., de Medeiros I.A., Martino H.S.D. Protein quality and free diets differentially impact energy intake, body composition, and hepatic lipid metabolism in growing rats. Nutrients, 2024, 16(11): 1754. https://doi.org/10.3390/nu16111754
Guillin F.M., Gaudichon C., Guérin-Deremaux L., Lefranc-Millot C., Azzout-Marniche D. Real ileal amino acid digestibility of pea protein compared to casein in healthy humans: A randomized trial. American Journal of Clinical Nutrition, 2021, 114(5): 1806-1817. https://doi.org/10.1093/ajcn/nqab234
Jeong H., Lee H., Han J.A., Lee S., Lee K.W. Effect of a modified casein:whey protein ratio in a low-protein diet on growth performance, BUN, and apparent digestibility of nursery pigs. Food Science of Animal Resources, 2021, 41(5): 825-838. https://doi.org/10.5851/kosfa.2021.e42
Khachatryan V., Mkrtchyan G., Abrahamyan L. Maintaining animal welfare in scientific research. Sechenov Medical Journal, 2021, 12(3): 4-13. https://doi.org/10.47093/2218-7332.2021.12.3.4-13
Kumar M., Tomar M., Punia S., Dhakane-Lad J., Dhumal S., Changan S., Senapathy M., Berwal M.K., Sampathrajan V., Sayed A.A.S., Rusu A.V., Corciovă A., Mekhemar M. Plant-based proteins and their multifaceted industrial applications. LWT – Food Science and Technology, 2022, 154(1): 112620. https://doi.org/10.1016/j.lwt.2021.112620
Lærke H.N, Theil P.K., Knudsen K.E.B. Enrichment of metabolic cages for rats: Effects on welfare and data quality. Journal of the American Association for Laboratory Animal Science, 2025, 64(1): 12-22. https://doi.org/10.30802/AALAS-JAALAS-24-000054
Liu L., Jiang X., Chen Y., Yaqoob S., Xiu L., Liu H., Zheng M., Cai D., Liu J. Germination-induced modifications of starch structure, flour-processing characteristics, and in vitro digestive properties in maize. Food Chemistry, 2024, 22(6): 101430. https://doi.org/10.1016/j.fochx.2024.101430
Lonnie M., Hooker E., Brunstrom J.M., Corfe B.M., Green M.A., Watson A.W., Williams E.A., Stevenson E.J., Penson S., Johnstone A.M. Protein for life: Optimal protein intake, sustainable sources, and appetite effects in ageing adults. Nutrients, 2018, 10(3): 360. https://doi.org/10.3390/nu10030360
Mulak D.G., Ahure D., Eke M.O. Production and quality evaluation of cookies from wheat–almond–carrot composite flour. International Journal of Food Science and Biotechnology, 2020, 5(4): 45-51. https://doi.org/10.11648/j.ijfsb.20200504.11
Musigwa S., Cozannet P., Asiamah C.A., Wu S.B. Effects of dietary protein levels, net energy levels, and essential amino acid-to-true protein ratios on broiler performance. Animals, 2024, 14(21): 3065. https://doi.org/10.3390/ani14213065
Nychyk O., Barton W., Rudolf A.M., Boscaini S., Walsh A., Bastiaanssen. Protein quality and quantity influence the effect of dietary fat on weight gain and tissue partitioning via host-microbiota changes. Cell Reports, 2021, 35(6): 109093. https://doi.org/10.1016/j.celrep.2021.109093
Olamiti G., Ramashia S.E. Impact of composite flour on nutritional, bioactive and sensory characteristics of pastry foods: A review. Current Research in Nutrition and Food Science, 2024, 12(3): 1018-1034. Available at: https://www.foodandnutritionjournal.org/pdf/vol12no3/Nutrition_Vol12_No3_p_1018-1034.pdf
Olanrewaju O.O., Fafiolu A.O., Oso A.O. Nutritive value of animal protein sources in rat model: A review. Nigerian Journal of Animal Production, 2024, 51(1): 123-134. https://doi.org/10.51791/njap.vi.7153
Otoo E., Asibuo J.Y., Akromah R. Enhancing protein and vitamin A intake in maize-based porridge using orange-fleshed sweet potato and cowpea composite flour. Legume Science, 2024, 6(2): e231. https://doi.org/10.1002/leg3.231
Panaite T.D., Olteanu M., Ropota M., Vasile G., Corbu A.R. Potential of peas as a sustainable protein source in pig diets: A review. Agriculture, 2025, 15(4): 789. https://doi.org/10.3390/agriculture15040789
Pinckaers P.J.M., Hendriks F.K., Hermans W.J.H., Senden J.M., Goessens J.P.B., Van Loon L.J.C. The muscle protein synthetic response to ingestion of plant-based protein blends does not differ from milk protein in young males. The Journal of Nutrition, 2021, 151(9): 2731-2740. https://doi.org/10.1093/jn/nxab195
Roschke N.N., Hillebrandt K.H., Polenz D., Klein O., Gassner J.M.G.V., Pratschke J., Krenzien F., Sauer IM., Raschzok N., Moor S. Optimizing environmental enrichment for Sprague Dawley rats: Exemplary insights into the liver proteome. PLOS ONE, 2024, 19(4): e0297497. https://doi.org/10.1371/journal.pone.0297497
Salles J., Guillet C., Le Bacquer O., Malnero-Fernandez C., Giraudet C., Patrac V., Berry A., Denis P., Pouyet C., Gueugneau M., Boirie Y., Jacobs H., Walrand S. Pea proteins have anabolic effects comparable to milk proteins on whole-body protein retention and muscle protein metabolism in old rats. Nutrients, 2021, 13(12): 4234. https://doi.org/10.3390/nu13124234
Schipper L., Oosting A., Scheurink A.J.W., van Dijk G., van der Beek E.M.A. customized grain-based diet as an alternative to the AIN-93 diet for mice. PLOS ONE, 2024, 19(3): e0298765. https://doi.org/10.1371/journal.pone.0298765
Sobotka W., Drażbo A. The effect of dietary protein restriction in phase feeding systems on nitrogen metabolism and excretion in pig production. Animals, 2025, 15(11): 1521. https://doi.org/10.3390/ani15111521
Terefe Z.K., Omwamba M., Nduko J. Effect of solid-state fermentation on nutritional and antinutritional properties of maize flour. Food Science & Nutrition, 2021, 9(9): 5221-5231. https://doi.org/10.1002/fsn3.2485
Tesalonika T., Sianipar O., Suryanto E. Advancements in rodent housing systems: Welfare and research implications. Livestock and Animal Research, 2025, 23(1): 45-56. https://doi.org/10.20961/lar.v22i3.84171
Udedeh H.E., Emelike N.J.T., Okorie-Humphrey C., Enyi C.U. Nutritional and sensory properties of cookies from African walnut (Tetracarpidium conophorum), Justicia carnea, and wheat flour. IPS Applied Journal of Nutrition, Food and Metabolism Science, 2024, 2(1): 19-26. https://doi.org/10.54117/iajnfms.v2i1.49
Warrilow A., Mellor D., McKune A.J., Pumpa K. Dietary fat, fibre, satiation, and satiety: A systematic review of acute studies. European Journal of Clinical Nutrition, 2019, 73(3): 333-344. https://doi.org/10.1038/s41430-018-0295
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How to Cite
ANYADIOHA, Josephat Ikechukwu et al.
Evaluation of protein quality and safety of high-fat cookies made from composite flours of hildegardia barteri, maize, and plantain using in vivo feeding trials.
Food Science and Applied Biotechnology, [S.l.], v. 9, n. 1, p. 209-220, mar. 2026.
ISSN 2603-3380.
Available at: <https://www.ijfsab.com/index.php/fsab/article/view/504>. Date accessed: 19 may 2026.
doi: https://doi.org/10.30721/fsab2026.v9.i1.504.