Evaluation of High-Protein Red Lentils in the Development of Gluten-Free Functional Bread

Evaluation of High-Protein Red Lentils in the Development of Gluten-Free Functional Bread | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Evaluation of High-Protein Red Lentils in the Development of Gluten-Free Functional Bread Büşra DOKUZ MURAT, Gülefşan PAŞA, Merve SAMANCI, Pelin ÇETİNKAYA, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7271202/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The increasing demand for nutritionally enhanced and gluten-free functional foods has catalyzed innovation in bread formulations targeting individuals with coeliac disease and health-conscious consumers. This study aimed to develop a gluten-free bread utilizing high-protein red lentil flour, optimized through a Box–Behnken experimental design. Lentil flour, buckwheat flour, and olive oil were selected as independent variables, while almonds, hazelnuts, pumpkin seeds, and spices were included at fixed levels to enhance nutritional and sensory properties. Fifteen formulations were evaluated through sensory analysis, and statistical optimization identified the ideal composition as 50 g lentil flour, 45 g buckwheat flour, and 6.5 g olive oil. The optimized bread exhibited superior sensory acceptability and a favorable nutritional profile, delivering 12.82 g protein, 8.07 g dietary fibre, 58.85 mg omega-3 fatty acids, and 0.52 mg vitamin E per 100 g, with gluten content well below the Codex Alimentarius threshold (< 20 ppm). The product was free from animal-derived ingredients and chemical additives, making it suitable for vegan and lactose-intolerant populations. Despite a relatively high fat content (11.77 g/100 g), the predominant fatty acid was oleic acid (64%), supporting cardiovascular health. These findings demonstrate the feasibility of producing a nutritionally dense, sensory-acceptable, and additive-free gluten-free bread using sustainable plant-based ingredients. By providing a scientifically optimized formulation that addresses key nutritional and sensory challenges in gluten-free baking, this study makes a meaningful contribution to the literature on functional food innovation, sustainable ingredient utilization, and inclusive dietary product development. Functional food gluten-free bread lentil flour Box-Behnken design Figures Figure 1 1. Introduction The global food industry is currently undergoing a period of sustained growth, driven by evolving consumer preferences for healthier and more functionally beneficial products. This transformation is primarily influenced by shifts in dietary patterns, particularly among health-conscious consumers who increasingly prioritize nutrient-dense products rich in protein, dietary fiber, and omega-3 fatty acids [ 1 ]. In response to these trends, plant-based and protein-fortified snack products have gained significant momentum, forming a dynamic and continuously evolving market segment that aligns with ethical consumption values and sustainability imperatives [ 2 ]. In line with sustainable production paradigms, increasing emphasis is being placed on the valorization of agricultural and food processing by-products. These efforts aim to reduce food waste and mitigate the environmental impacts associated with conventional snack production, while simultaneously enhancing the nutritional profile of the final product [ 3 ]. Gluten-free snacks, once limited to individuals with celiac disease, have transitioned into the mainstream, now appealing to a broader demographic of health-oriented consumers. This development is particularly evident in North American and European markets, where demand for gluten-free alternatives is contributing substantially to market expansion [ 4 ]. Gluten, identified as the primary trigger of clinical manifestations such as diarrhea, constipation, abdominal bloating, nausea, and vomiting in individuals with coeliac disease, is a composite of water-insoluble prolamin proteins that constitutes a major storage protein in cereal grains [ 5 , 6 ]. Cereals including wheat, barley, rye, and, to a lesser extent, oats contain gluten, whereas legumes and grains such as lentils, rice, and corn are naturally gluten-free [ 7 ]. Among gluten-free food sources, lentils are particularly noteworthy due to their high protein content (approximately 25–30%) and their richness in essential amino acids [ 8 ]. Furthermore, lentils are considered a valuable source of prebiotic carbohydrates, attributed to their significant concentration of oligosaccharides. In addition to their protein and carbohydrate content, lentils also provide substantial amounts of dietary fibre, phosphorus, vitamin B9 (folate), and thiamine, thereby contributing to their nutritional and functional value in gluten-free diets [ 9 ]. Bread, a staple food product traditionally made from wheat flour, is widely consumed in daily diets across various populations. Although a range of gluten-free alternatives -including bread products- has been developed using diverse raw materials to meet the dietary needs of individuals with coeliac disease, these products often suffer from suboptimal sensory, textural, and nutritional qualities. A significant formulation challenge lies in the complete replacement of gluten, as this protein plays a crucial role in conferring the viscoelastic and structural characteristics essential for high-quality breadmaking [ 10 ]. Findings from a randomized controlled trial have indicated that individuals diagnosed with coeliac disease frequently avoid bread consumption. Contributing factors include limited product diversity, restricted market and bakery availability, dissatisfaction with flavor and texture, and difficulties associated with mastication. The insufficient variety of gluten-free breads and the low consumer acceptability of existing commercial options have underscored the need for innovation in this area [ 11 ]. Accordingly, the present study aimed to develop a gluten-free bread with enhanced nutritional value, sensory appeal, and accessibility, utilizing lentil flour as the primary ingredient. 2. Materials And Methods 2.1. Materials used The formulation developed in this study comprised organic red lentil flour, certified gluten-free buckwheat flour, almonds, hazelnuts, pumpkin seeds, ground coriander, fennel seeds, and fresh yeast. All raw materials were procured from certified local organic suppliers to ensure quality and traceability. The ingredients were stored in vacuum-sealed packaging under controlled conditions, protected from light and moisture, at a temperature of + 4°C to preserve their physicochemical and microbiological integrity prior to use. 2.2. Experimental design Based on the outcomes of preliminary trials, minimum and maximum levels were established for red lentil flour (10–90 g), buckwheat flour (10–90 g), and olive oil (3–10 g). As the initially fixed amount of yeast did not yield satisfactory results, yeast was incorporated into the formulation at a rate equivalent to 4% of the total flour weight. Hazelnuts, almonds, and pumpkin seeds were each included at a level of 5%, while coriander and fennel were added at 2.5%. In the experimental design, the quantities of red lentil flour, buckwheat flour, and olive oil were treated as independent variables. A Box-Behnken design was employed to optimize these three factors using the Minitab statistical software, with the levels of oilseeds, yeast, and spices held constant throughout the experiments. 2.3. Production process All ingredients were thoroughly mixed to achieve a homogeneous dough, which was subsequently kneaded for 10 minutes. The dough was then allowed to rest for 30 minutes at ambient temperature, followed by a pre-fermentation period of 1 hour and 30 minutes at 35°C. Baking was carried out at 180°C for approximately 35 minutes. Upon completion of baking, the breads were cooled at room temperature for one hour prior to conducting sensory evaluations. 2.4. Sensory assessment Each bread sample was subjected to sensory evaluation by a panel of 22 semi-trained assessors at Avrasya University. A 9-point hedonic scale was employed for the assessment, ranging from 1 (“dislike extremely”) to 9 (“like extremely”). The sensory attributes evaluated included flavour, swallowability, colour, odour, chewability, and overall acceptability. 2.5. Chemical analysis The chemical composition of the formulation that received the highest overall acceptability score, as determined through optimization, was subsequently analyzed. These analyses were conducted at the laboratories of the Scientific and Technological Research Council of Türkiye, Marmara Research Centre (TÜBİTAK MAM). 2.6. Statistical analysis A Box–Behnken experimental design was employed to investigate the effects of selected process parameters on the structural attributes of the product. Three independent variables were considered: lentil flour content (𝑥₁), buckwheat flour content (𝑥₂), and oil content (𝑥₃). Each variable was examined at three coded levels: low (− 1), medium (0), and high (+ 1). The experimental data were analyzed using response surface methodology (RSM), with all computations conducted via Minitab 17 software. To determine the statistical significance of the model and individual effects, analysis of variance (ANOVA) was performed at a confidence level of 95% (p < 0.05). Response surfaces were generated to visualize the interaction effects between variables by holding one variable constant at its center point while varying the other two across their coded levels. 3. Results And Discussion This study evaluated the sensory acceptance of a newly developed gluten-free functional bread through 15 distinct formulations, designed using the Box–Behnken experimental design (Table 1 ). Lentil flour, buckwheat flour, and olive oil were selected as independent variables, while almonds, hazelnuts, pumpkin seeds, coriander, and fennel were included at fixed levels as enriching components (Fig. 1 ). Table 1 Box-Behnken experimental design for bread processing * Run Order (RO) Lentil Flour (g) Buckwheat Flour (g) Oil (g) 1 50 50 6,5 2 10 10 6,5 3 50 90 10 4 90 90 6,5 5 50 10 10 6 50 50 6,5 7 50 50 6,5 8 50 10 3 9 10 50 3 10 90 10 6,5 11 10 90 6,5 12 50 90 3 13 90 50 10 14 10 50 10 15 90 50 3 * Source: Authors own work The sensory analysis revealed that formulations containing 50 g each of lentil and buckwheat flours achieved the highest scores across all sensory parameters ( Supplementary 1 ). Statistical evaluation demonstrated that lentil flour (LF), buckwheat flour (BF), and oil content significantly affected overall acceptability (p < 0.001), as well as all other sensory characteristics (Table 2 ). The response surface methodology (RSM) confirmed significant linear, square, and interaction effects, indicating a strong influence of the three primary ingredients on the sensory attributes of the final product. Table 2 P-values for the results of the response surface analysis for all variables * Source Flavour Chewiness Color Odor Swallowability Overall Acceptability Total Model < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 Linear < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 LF < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 BF < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 Oil < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 Square < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 LF*LF < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 BF*BF < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 Oil*Oil < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 2 -Way Interaction < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 LF*BF < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 < 0,001 LF*Oil 0,676 0,44 0,009 0,214 0,539 0,665 0,607 BF*Oil 0,013 0,007 0,917 < 0,001 0,015 0,031 < 0,001 ( LF: Lentil Flour, BF: Buckwheat flour, *Source: Authors own work) The optimal formulation determined by RSM optimization using Minitab software is detailed in Table 3 . This optimized product comprised 50 g lentil flour, 45 g buckwheat flour, and 6.5 g olive oil, enriched with 5 g each of almond and pumpkin seeds, 2.5 g fennel, 2.5 g cumin powder, and 200 mL water, along with 4% yeast. Table 3 Final bread formulation* Ingredients LF BF Oil AP PS Yeast Fennel Cumin Powder Water Amount (g) 50 g 45 g 6,5 g 5 g 5 g %4 2,5 g 2,5 g 200 mL ( LF: Lentil Flour, BF: Buckwheat flour, AP: Almond flour, PS: Pumpkin seed, *Source: Authors own work) Compositional analysis of the final product (per 100 g) revealed 31.6 g carbohydrate, 12.8 g protein, and 11.7 g fat, of which 64% was oleic acid. The product also provided 58.85 mg omega-3 fatty acids, 8.07 g dietary fibre, and 0.52 mg α-tocopherol (vitamin E). Gluten content was measured at 7.89 ppm, well below the Codex Alimentarius Commission threshold of 20 ppm, thus qualifying the product as gluten-free [ 12 ]. These values are presented in Table 4 , alongside their respective contributions to daily intake based on TUBER 2022 and WHO guidelines. Table 4 Nutrient Content of Gluten-Free Functional Bread (per 100 g) and Its % Contribution to Daily Requirements According to TUBER 2022 and WHO Nutrient Gluten Free Bread Per (100g) % Daily Value (TÜBER 2022) % Daily Value (WHO) Energy (kcal/100 g) 300 kkal/100g 15 15 Carbohydrate (g/100 g) 31,64 g 10.5 12.2 Protein (g/100 g) 12,82 g 25.6 27.9 Fat (g/100 g) 11,77 g 18.1 16.8 Oleic acid (%) %64,42 NV NV Omega-3 (mg/100 g)¹ 58,85 mg/100g 23.5 11.7 Dietary fibre (g/100 g) 8,07 g 32.3 32.3 Vitamin E (α-tocopherol, mg) 0.52 mg 4.3 3.5 Gluten (ppm) 7,89 ppm - - Moisture (g/100 g) 33,36g/100g - - Ash (g/100 g) 2,34 g/100g - - (* The table presents the macronutrient and micronutrient composition of the developed gluten-free functional bread per 100 g and its contribution to the daily requirements of an adult female with a 2000 kcal energy need, based on TUBER 2022 and WHO reference values. There is no specific recommended daily intake for oleic acid set by TUBER or WHO. ¹Omega-3 content contribution was calculated according to Report of the sub-committee on recommendations for intake of polyunsaturated fatty acids in healthy adults [Internet]. Washington, DC: International Society for the Study of Fatty Acids and Lipids; 2004. Available from : http://www.issfal.org/news-links/resources/publications/PUFAIntakeReccomdFinalReport.pdf , Source: Authors’ own work.) The nutritional profile of this bread addresses a key challenge in gluten-free diets: ensuring sufficient intake of essential macro- and micronutrients. Previous studies have reported that between 20–38% of individuals adhering to gluten-free diets, particularly those with coeliac disease, suffer from nutritional deficiencies including energy, protein, fiber, vitamins, and minerals [ 13 , 14 ]. Many commercially available gluten-free products are typically high in sugar, fat, and sodium while being low in dietary fibre and essential vitamins [ 15 ]. Thus, the current formulation offers a promising nutritional alternative. In the present study, buckwheat and lentil flours served as the principal flours. Buckwheat is a gluten-free pseudocereal with a high lysine content and dietary fibre [ 16 ]. Lentil flour is rich in high-quality plant-based protein, minerals, and prebiotic carbohydrates [ 17 , 18 ]. Unlike earlier formulations that use lentil flour in minimal proportions, this study successfully incorporated it at a 50% level, resulting in a protein content of 12.82 g/100 g substantially higher than in conventional gluten-free breads [ 19 ]. The absence of animal-derived ingredients such as eggs or milk powder renders the product suitable for vegans and individuals with lactose intolerance. Furthermore, the inclusion of almonds and hazelnuts, both rich in oleic acid and α-tocopherol, contributed to the product’s healthy fat profile and antioxidant content. These nuts are known for enhancing cardiovascular health and providing vitamin E [ 20 , 21 ], and their presence may also positively influence product shelf life due to antioxidant activity [ 22 ]. The total vitamin E content (0.52 mg/100 g) aligns with values expected from nut-enriched formulations [ 23 ]. Although the fat content (11.77 g/100 g) is relatively high, the predominant fatty acid is oleic acid, which supports heart health [ 24 ]. The omega-3 content, at 58.85 mg/100 g, is notable, especially when compared with traditional gluten-free products [ 25 ] and contributes to cardiovascular and neurological health [ 26 ]. This quantity meets approximately 11.7% of the daily recommended intake for healthy adults [ 27 ]. Moreover, the product contains 2,459 mg of omega-6 fatty acids per 100 g, contributing appropriately to the recommended intake range of 11–22 g/day [ 28 ]. However, the omega-6/omega-3 ratio was calculated as 41.8:1, which exceeds the ideal range of 4:1 to 5:1 [ 27 ] likely due to the absence of omega-3-rich sources such as chia or flaxseed. Future research should consider reformulating to achieve a more favourable fatty acid ratio. The carbohydrate content (31.64 g/100 g) is lower than that of many commercial gluten-free breads [ 29 , 30 ]. Since 70% of the product’s caloric value is derived from carbohydrates, it presents a balanced glycaemic profile. The inclusion of legume flour has been shown to mitigate postprandial glycaemic response and enhance satiety [ 31 ] suggesting potential benefits for individuals with metabolic disorders. The fibre content (8.07 g/100 g) exceeds that of most gluten-free commercial products, which typically range from 3–6% [ 32 ], fulfilling approximately 25% of the recommended daily fibre intake. Dietary fibre contributes to intestinal health, glycaemic regulation, cholesterol reduction, and prolonged satiety, and also improves the shelf life of baked products through structural stabilization. Moisture and ash contents were determined to be 33.36% and 2.34 g/100 g, respectively values consistent with gluten-free bread literature [ 33 ]. The product maintained optimal chewability without requiring texture-modifying additives, indicating a naturally stable consistency. Sensory analysis confirmed high consumer acceptance for all evaluated parameters, including appearance, aroma, flavour, and mouthfeel. The contribution of nuts to chewability and the enhancement of flavour through coriander and fennel were particularly notable. Similar findings have been reported by [ 33 ], who observed increased sensory scores in gluten-free breads containing spices. Likewise, studies by Tunç ve Kahyaoglu (2016) and Oliete et al. (2008) demonstrated improved texture and consumer acceptability with the inclusion of nut pastes [ 34 , 35 ]. Overall, a gluten-free bread with a balanced nutritional profile and high sensory acceptance was successfully developed without the use of chemical additives, thickeners, or preservatives. This formulation is not only suitable for individuals with coeliac disease but also for those following plant-based or low glycaemic index diets. Furthermore, the product’s concentrated nutrient content and shelf-stability potential make it an appropriate option for crisis and emergency food supply contexts. The use of the Box–Behnken design allowed efficient exploration of the formulation space with a relatively small sample size, enabling time- and cost-effective optimization. However, the study has several limitations. Firstly, the glycaemic index was estimated theoretically rather than clinically determined. Secondly, comprehensive evaluations such as shelf life, microbial safety, textural properties, and physicochemical stability were not conducted. Thirdly, the sensory panel included only a limited number of healthy individuals, without feedback from coeliac patients or those routinely following a gluten-free diet. Additionally, consumer surveys and market preference data were not collected. Future research should address these limitations by conducting in vivo glycaemic assessments, stability analyses, and consumer-based preference studies across diverse demographics. Tailored product development targeting specific populations, such as children, older adults, or athletes, would further enhance the market relevance of gluten-free functional breads. 4. Conclusion This study successfully developed a nutritionally enriched, gluten-free functional bread using high-protein red lentil flour as the primary ingredient, optimized through a Box–Behnken experimental design. The resulting formulation exhibited superior sensory properties and a well-balanced nutritional profile, characterized by elevated levels of protein, dietary fibre, and beneficial unsaturated fatty acids, particularly oleic acid. The product also demonstrated compliance with international gluten-free standards, rendering it suitable for individuals with coeliac disease, as well as for those adhering to plant-based or health-conscious diets. The compositional analysis revealed that the optimized formulation addressed several common nutritional deficiencies observed in gluten-free diets, particularly in terms of protein, fibre, and essential fatty acid intake. The successful incorporation of 50% lentil flour, a higher proportion than typically employed, contributed significantly to the product’s protein density without compromising sensory acceptability. Additionally, the inclusion of almonds, hazelnuts, and spices enhanced the organoleptic and antioxidant characteristics of the bread, contributing to its favourable reception in sensory evaluations. Importantly, this study demonstrated the feasibility of producing additive-free, plant-based gluten-free bread with functional and nutritional advantages using sustainable and regionally available raw materials. The use of response surface methodology facilitated a robust and efficient optimization process, enabling the identification of an optimal ingredient matrix within a limited number of trials. Despite these promising outcomes, the study presents limitations that warrant further investigation. Clinical validation of the glycaemic index, shelf-life assessment, microbial safety analysis, and broader sensory evaluations, particularly among coeliac patients and habitual gluten-free consumers, are necessary to establish the product’s market readiness and health claims. Furthermore, future studies should explore strategies to improve the omega-6/omega-3 ratio through the inclusion of additional omega-3-rich ingredients, as well as expand the application of this formulation to diverse population groups, including pediatric, geriatric, and athletic cohorts. The findings underscore the potential of lentil-based formulations as a viable alternative in the gluten-free product market, offering both nutritional and sensory benefits. This research contributes to the advancement of functional food innovation by addressing the dual imperatives of dietary health and sustainable food development. Declarations Author Contribution B.D.M. conceived and designed the study, developed the research methodology, and carried out the formal analysis. She led the investigation, coordinated resources, curated and managed the data, prepared the original draft of the manuscript, created the visualizations, and provided overall supervision of the research process.G.P., P.Ç., and A.A. were responsible for the preparation of bread samples and the execution of sensory evaluations. They contributed to the acquisition of experimental data and participated in the interpretation of the results.M.S. contributed substantially to the writing, critical review, and editing of the manuscript, ensuring clarity, coherence, and alignment with journal requirements. Acknowledgement AcknowledgementThis study was supported by the TÜBİTAK 2209-A programme. We would like to thank the Scientific and Technological Research Council of Turkey (TÜBİTAK). Our sincere thanks also go to all the panellists who contributed to the product development and analysis proces-ses, as well as to the academic staff of the Departments of Nutrition and Dietetics and Gastro-nomy and Culinary Arts at Avrasya University. The authors wish to express their sincere gratitude to Assistant Professor Abdurrahman Atalay and Sefa Murat Dokuz for their valuable assistance in the English language editing of this manuscript.A preliminary version of this study was presented as an oral presentation at the Sustainable Nutrition Days: Shaping the Future with Turkish Cuisine symposium (Sürdürülebilir Beslenme Günleri: Türk Mutfağı ile Geleceğe Yön Veriyoruz Sempozyumu-SÜRBES2025) held in Turkey. The presentation was not published in any proceedings or academic journals. Data Availability All data supporting the findings of this study are included within the main text and its Supplementary Information. 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Med. , c. 233, sy 6, ss. 674-688, Haz. 2008, doi: 10.3181/0711-MR-311. ISSFAL, “Intake of PUFA in Healthy Adults”, International Society for the Study of Fatty Acids and Lipids. Erişim: 30 Temmuz 2025. [Çevrimiçi]. Erişim adresi: https://www.issfal.org/statement-3 European Food Safety Authority, “Scientific Opinion on the substantiation of health claims related to linoleic acid and maintenance of normal blood cholesterol concentrations (ID 489) pursuant to Article 13(1) of Regulation (EC) No 1924/2006”, EFSA J. , sy 2009; 7(9):1276, doi: 10.2903/j.efsa.2009.1276. B. I. Achalu, H. A. Tessema, ve A. T. Ayele, “Development and characterization of gluten-free bread enriched with whey protein from sorghum-rice composite flours”, Appl. Food Res. , c. 5, sy 2, s. 101133, Ara. 2025, doi: 10.1016/j.afres.2025.101133. Z. Šmídová ve J. Rysová, “Gluten-Free Bread and Bakery Products Technology”, Foods , c. 11, sy 3, s. 480, Şub. 2022, doi: 10.3390/foods11030480. P. Binou, A. E. Yanni, ve V. T. Karathanos, “Physical properties, sensory acceptance, postprandial glycemic response, and satiety of cereal based foods enriched with legume flours: a review”, Crit. Rev. Food Sci. Nutr. , c. 62, sy 10, ss. 2722-2740, Nis. 2022, doi: 10.1080/10408398.2020.1858020. M. Cornicelli, M. Saba, N. Machello, M. Silano, ve S. Neuhold, “Nutritional composition of gluten-free food versus regular food sold in the Italian market”, Dig. Liver Dis. , c. 50, sy 12, ss. 1305-1308, Ara. 2018, doi: 10.1016/j.dld.2018.04.028. L. Ruiz-Aceituno, N. Casado, E. Arriero-Romo, S. Morante-Zarcero, A. Lázaro, ve I. Sierra, “Development of Gluten-Free Bread Based on Maize and Buckwheat and Enriched with Aromatic Herbs and Spices”, Appl. Sci. , c. 14, sy 8, s. 3348, Nis. 2024, doi: 10.3390/app14083348. M. T. Tunç ve T. Kahyaoglu, “Improving Rheological and Baking Properties of Gluten-Free Breads Using Defatted Hazelnut Flour with Various Gums”, Int. J. Food Eng. , c. 12, sy 4, ss. 343-353, Haz. 2016, doi: 10.1515/ijfe-2015-0207. B. Oliete, M. Gómez, V. Pando, E. Fernández-Fernández, P. A. Caballero, ve F. Ronda, “Effect of Nut Paste Enrichment on Physical Characteristics and Consumer Acceptability of Bread”, Food Sci. Technol. Int. , c. 14, sy 3, ss. 259-269, Haz. 2008, doi: 10.1177/1082013208095691. Additional Declarations No competing interests reported. Supplementary Files Supplementarymaterial.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7271202","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":497800052,"identity":"83d892c3-5a76-478b-be4d-33d6abd19e15","order_by":0,"name":"Büşra DOKUZ MURAT","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEUlEQVRIiWNgGAWjYBACAyQ24wMEm40oLczMSBygFh4itLBJEKXFXCL92YcfDNsSt7efP1bxs+2evPyM5AcMH8oOM9hLH8CqxXJGjvHMHobbiXPOJLPd7G0rNtxwI82Acca5www8fAnYHXYjhxnohNuJMxiS2W7wnElg3CCRYMDM2wbUgsNlBjfSHzP+AWnhf8xW+OdMgv38GekfmP/i1ZJgzAy2RSKZjZmnIiGx4UaOATMjHi2WPW+MmWUMbhvPkHhsLC1TkZC84cybgoM959J5eM7gCDF2oMPeVNyWncGf+PDjG4ME2/nt6Rsf/CizlmPvwa4F6jxkjkACwwEG3NGCDfAfIEHxKBgFo2AUjAQAAMTbWoRmm3clAAAAAElFTkSuQmCC","orcid":"","institution":"Avrasya University","correspondingAuthor":true,"prefix":"","firstName":"Büşra","middleName":"DOKUZ","lastName":"MURAT","suffix":""},{"id":497800053,"identity":"5311a539-eba0-41bc-ad42-24df42744bb9","order_by":1,"name":"Gülefşan PAŞA","email":"","orcid":"","institution":"Avrasya University","correspondingAuthor":false,"prefix":"","firstName":"Gülefşan","middleName":"","lastName":"PAŞA","suffix":""},{"id":497800054,"identity":"db20dbbb-8a5c-4398-9dff-fa9e76f071ae","order_by":2,"name":"Merve SAMANCI","email":"","orcid":"","institution":"Avrasya University","correspondingAuthor":false,"prefix":"","firstName":"Merve","middleName":"","lastName":"SAMANCI","suffix":""},{"id":497800055,"identity":"a799f2a2-5231-4192-9aae-f57da98806a4","order_by":3,"name":"Pelin ÇETİNKAYA","email":"","orcid":"","institution":"Avrasya University","correspondingAuthor":false,"prefix":"","firstName":"Pelin","middleName":"","lastName":"ÇETİNKAYA","suffix":""},{"id":497800056,"identity":"f687aecd-cd0f-49f3-915d-b2f7f8322faf","order_by":4,"name":"Miray DOĞAN","email":"","orcid":"","institution":"Avrasya University","correspondingAuthor":false,"prefix":"","firstName":"Miray","middleName":"","lastName":"DOĞAN","suffix":""},{"id":497800057,"identity":"44c3fb41-1792-4828-944d-54ebfc68c890","order_by":5,"name":"Asuman AHMETBEYOĞLU","email":"","orcid":"","institution":"Avrasya University","correspondingAuthor":false,"prefix":"","firstName":"Asuman","middleName":"","lastName":"AHMETBEYOĞLU","suffix":""}],"badges":[],"createdAt":"2025-08-01 12:23:28","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7271202/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7271202/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":88760540,"identity":"a21dcf2e-f064-4f8e-b107-4ea855efd763","added_by":"auto","created_at":"2025-08-11 07:54:36","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":366536,"visible":true,"origin":"","legend":"\u003cp\u003eBread manufacturing process\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7271202/v1/5ee896a9726fcbf7f1e43b7a.png"},{"id":89063266,"identity":"da3db4c3-ea2d-47c7-81f0-e39381505eef","added_by":"auto","created_at":"2025-08-14 09:58:50","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1270370,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7271202/v1/2ead1d06-c6d5-45ac-9751-bd98f758e06b.pdf"},{"id":88759641,"identity":"49043a07-55aa-45ef-85c2-11ce2b111631","added_by":"auto","created_at":"2025-08-11 07:46:36","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":7287272,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-7271202/v1/4e74f08c4cbcd71f7bf1f13e.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Evaluation of High-Protein Red Lentils in the Development of Gluten-Free Functional Bread","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe global food industry is currently undergoing a period of sustained growth, driven by evolving consumer preferences for healthier and more functionally beneficial products. This transformation is primarily influenced by shifts in dietary patterns, particularly among health-conscious consumers who increasingly prioritize nutrient-dense products rich in protein, dietary fiber, and omega-3 fatty acids [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. In response to these trends, plant-based and protein-fortified snack products have gained significant momentum, forming a dynamic and continuously evolving market segment that aligns with ethical consumption values and sustainability imperatives [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. In line with sustainable production paradigms, increasing emphasis is being placed on the valorization of agricultural and food processing by-products. These efforts aim to reduce food waste and mitigate the environmental impacts associated with conventional snack production, while simultaneously enhancing the nutritional profile of the final product [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eGluten-free snacks, once limited to individuals with celiac disease, have transitioned into the mainstream, now appealing to a broader demographic of health-oriented consumers. This development is particularly evident in North American and European markets, where demand for gluten-free alternatives is contributing substantially to market expansion [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eGluten, identified as the primary trigger of clinical manifestations such as diarrhea, constipation, abdominal bloating, nausea, and vomiting in individuals with coeliac disease, is a composite of water-insoluble prolamin proteins that constitutes a major storage protein in cereal grains [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eCereals including wheat, barley, rye, and, to a lesser extent, oats contain gluten, whereas legumes and grains such as lentils, rice, and corn are naturally gluten-free [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAmong gluten-free food sources, lentils are particularly noteworthy due to their high protein content (approximately 25\u0026ndash;30%) and their richness in essential amino acids [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Furthermore, lentils are considered a valuable source of prebiotic carbohydrates, attributed to their significant concentration of oligosaccharides. In addition to their protein and carbohydrate content, lentils also provide substantial amounts of dietary fibre, phosphorus, vitamin B9 (folate), and thiamine, thereby contributing to their nutritional and functional value in gluten-free diets [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eBread, a staple food product traditionally made from wheat flour, is widely consumed in daily diets across various populations. Although a range of gluten-free alternatives -including bread products- has been developed using diverse raw materials to meet the dietary needs of individuals with coeliac disease, these products often suffer from suboptimal sensory, textural, and nutritional qualities. A significant formulation challenge lies in the complete replacement of gluten, as this protein plays a crucial role in conferring the viscoelastic and structural characteristics essential for high-quality breadmaking [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eFindings from a randomized controlled trial have indicated that individuals diagnosed with coeliac disease frequently avoid bread consumption. Contributing factors include limited product diversity, restricted market and bakery availability, dissatisfaction with flavor and texture, and difficulties associated with mastication. The insufficient variety of gluten-free breads and the low consumer acceptability of existing commercial options have underscored the need for innovation in this area [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Accordingly, the present study aimed to develop a gluten-free bread with enhanced nutritional value, sensory appeal, and accessibility, utilizing lentil flour as the primary ingredient.\u003c/p\u003e"},{"header":"2. Materials And Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1. Materials used\u003c/h2\u003e\u003cp\u003eThe formulation developed in this study comprised organic red lentil flour, certified gluten-free buckwheat flour, almonds, hazelnuts, pumpkin seeds, ground coriander, fennel seeds, and fresh yeast. All raw materials were procured from certified local organic suppliers to ensure quality and traceability. The ingredients were stored in vacuum-sealed packaging under controlled conditions, protected from light and moisture, at a temperature of +\u0026thinsp;4\u0026deg;C to preserve their physicochemical and microbiological integrity prior to use.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2. Experimental design\u003c/h2\u003e\u003cp\u003eBased on the outcomes of preliminary trials, minimum and maximum levels were established for red lentil flour (10\u0026ndash;90 g), buckwheat flour (10\u0026ndash;90 g), and olive oil (3\u0026ndash;10 g). As the initially fixed amount of yeast did not yield satisfactory results, yeast was incorporated into the formulation at a rate equivalent to 4% of the total flour weight. Hazelnuts, almonds, and pumpkin seeds were each included at a level of 5%, while coriander and fennel were added at 2.5%. In the experimental design, the quantities of red lentil flour, buckwheat flour, and olive oil were treated as independent variables. A Box-Behnken design was employed to optimize these three factors using the Minitab statistical software, with the levels of oilseeds, yeast, and spices held constant throughout the experiments.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3. Production process\u003c/h2\u003e\u003cp\u003eAll ingredients were thoroughly mixed to achieve a homogeneous dough, which was subsequently kneaded for 10 minutes. The dough was then allowed to rest for 30 minutes at ambient temperature, followed by a pre-fermentation period of 1 hour and 30 minutes at 35\u0026deg;C. Baking was carried out at 180\u0026deg;C for approximately 35 minutes. Upon completion of baking, the breads were cooled at room temperature for one hour prior to conducting sensory evaluations.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4. Sensory assessment\u003c/h2\u003e\u003cp\u003eEach bread sample was subjected to sensory evaluation by a panel of 22 semi-trained assessors at Avrasya University. A 9-point hedonic scale was employed for the assessment, ranging from 1 (\u0026ldquo;dislike extremely\u0026rdquo;) to 9 (\u0026ldquo;like extremely\u0026rdquo;). The sensory attributes evaluated included flavour, swallowability, colour, odour, chewability, and overall acceptability.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5. Chemical analysis\u003c/h2\u003e\u003cp\u003eThe chemical composition of the formulation that received the highest overall acceptability score, as determined through optimization, was subsequently analyzed. These analyses were conducted at the laboratories of the Scientific and Technological Research Council of T\u0026uuml;rkiye, Marmara Research Centre (T\u0026Uuml;BİTAK MAM).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003e2.6. Statistical analysis\u003c/h2\u003e\u003cp\u003eA Box\u0026ndash;Behnken experimental design was employed to investigate the effects of selected process parameters on the structural attributes of the product. Three independent variables were considered: lentil flour content (\u0026#119909;₁), buckwheat flour content (\u0026#119909;₂), and oil content (\u0026#119909;₃). Each variable was examined at three coded levels: low (\u0026minus;\u0026thinsp;1), medium (0), and high (+\u0026thinsp;1). The experimental data were analyzed using response surface methodology (RSM), with all computations conducted via Minitab 17 software. To determine the statistical significance of the model and individual effects, analysis of variance (ANOVA) was performed at a confidence level of 95% (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Response surfaces were generated to visualize the interaction effects between variables by holding one variable constant at its center point while varying the other two across their coded levels.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results And Discussion","content":"\u003cp\u003eThis study evaluated the sensory acceptance of a newly developed gluten-free functional bread through 15 distinct formulations, designed using the Box\u0026ndash;Behnken experimental design (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Lentil flour, buckwheat flour, and olive oil were selected as independent variables, while almonds, hazelnuts, pumpkin seeds, coriander, and fennel were included at fixed levels as enriching components (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBox-Behnken experimental design for bread processing\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRun Order (RO)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLentil Flour (g)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBuckwheat Flour (g)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eOil (g)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6,5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6,5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6,5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6,5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6,5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6,5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6,5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003e*\u003cem\u003eSource: Authors own work\u003c/em\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe sensory analysis revealed that formulations containing 50 g each of lentil and buckwheat flours achieved the highest scores across all sensory parameters (\u003cb\u003eSupplementary 1\u003c/b\u003e). Statistical evaluation demonstrated that lentil flour (LF), buckwheat flour (BF), and oil content significantly affected overall acceptability (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), as well as all other sensory characteristics (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The response surface methodology (RSM) confirmed significant linear, square, and interaction effects, indicating a strong influence of the three primary ingredients on the sensory attributes of the final product.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eP-values for the results of the response surface analysis for all variables\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"8\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSource\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFlavour\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eChewiness\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eColor\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eOdor\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eSwallowability\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eOverall Acceptability\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eTotal\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eModel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLinear\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLF\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBF\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOil\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSquare\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLF*LF\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBF*BF\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOil*Oil\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2 -Way Interaction\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLF*BF\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLF*Oil\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0,676\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0,44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e0,009\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0,214\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0,539\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0,665\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0,607\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBF*Oil\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003e0,013\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e0,007\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0,917\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e0,015\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e0,031\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0,001\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"8\"\u003e(\u003cem\u003eLF: Lentil Flour, BF: Buckwheat flour, *Source: Authors own work)\u003c/em\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eThe optimal formulation determined by RSM optimization using Minitab software is detailed in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. This optimized product comprised 50 g lentil flour, 45 g buckwheat flour, and 6.5 g olive oil, enriched with 5 g each of almond and pumpkin seeds, 2.5 g fennel, 2.5 g cumin powder, and 200 mL water, along with 4% yeast.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eFinal bread formulation*\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"10\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eIngredients\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLF\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBF\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eOil\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eAP\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003ePS\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eYeast\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eFennel\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eCumin Powder\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003eWater\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAmount (g)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e50 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e45 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,5 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e5 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e%4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2,5 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e2,5 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e200 mL\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"10\"\u003e(\u003cem\u003eLF: Lentil Flour, BF: Buckwheat flour, AP: Almond flour, PS: Pumpkin seed, *Source: Authors own work)\u003c/em\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eCompositional analysis of the final product (per 100 g) revealed 31.6 g carbohydrate, 12.8 g protein, and 11.7 g fat, of which 64% was oleic acid. The product also provided 58.85 mg omega-3 fatty acids, 8.07 g dietary fibre, and 0.52 mg α-tocopherol (vitamin E). Gluten content was measured at 7.89 ppm, well below the Codex Alimentarius Commission threshold of 20 ppm, thus qualifying the product as gluten-free [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. These values are presented in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, alongside their respective contributions to daily intake based on TUBER 2022 and WHO guidelines.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eNutrient Content of Gluten-Free Functional Bread (per 100 g) and Its % Contribution to Daily Requirements According to TUBER 2022 and WHO\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNutrient\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGluten Free Bread Per (100g)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e% Daily Value (T\u0026Uuml;BER 2022)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e% Daily Value (WHO)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eEnergy (kcal/100 g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e300 kkal/100g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCarbohydrate (g/100 g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e31,64 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eProtein (g/100 g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12,82 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e25.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e27.9\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFat (g/100 g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11,77 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e18.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e16.8\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOleic acid (%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e%64,42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNV\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOmega-3 (mg/100 g)\u0026sup1;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e58,85 mg/100g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e23.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e11.7\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDietary fibre (g/100 g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8,07 g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e32.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e32.3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eVitamin E (α-tocopherol, mg)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.52 mg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGluten (ppm)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7,89 ppm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMoisture (g/100 g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e33,36g/100g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAsh (g/100 g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2,34 g/100g\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e(*\u003cem\u003eThe table presents the macronutrient and micronutrient composition of the developed gluten-free functional bread per 100 g and its contribution to the daily requirements of an adult female with a 2000 kcal energy need, based on TUBER 2022 and WHO reference values. There is no specific recommended daily intake for oleic acid set by TUBER or WHO. \u0026sup1;Omega-3 content contribution was calculated according to Report of the sub-committee on recommendations for intake of polyunsaturated fatty acids in healthy adults [Internet]. Washington, DC: International Society for the Study of Fatty Acids and Lipids; 2004. Available from\u003c/em\u003e: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.issfal.org/news-links/resources/publications/PUFAIntakeReccomdFinalReport.pdf\u003c/span\u003e\u003cspan address=\"http://www.issfal.org/news-links/resources/publications/PUFAIntakeReccomdFinalReport.pdf\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e, \u003cem\u003eSource: Authors\u0026rsquo; own work.)\u003c/em\u003e\u003c/p\u003e\u003cp\u003eThe nutritional profile of this bread addresses a key challenge in gluten-free diets: ensuring sufficient intake of essential macro- and micronutrients. Previous studies have reported that between 20\u0026ndash;38% of individuals adhering to gluten-free diets, particularly those with coeliac disease, suffer from nutritional deficiencies including energy, protein, fiber, vitamins, and minerals [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Many commercially available gluten-free products are typically high in sugar, fat, and sodium while being low in dietary fibre and essential vitamins [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Thus, the current formulation offers a promising nutritional alternative.\u003c/p\u003e\u003cp\u003eIn the present study, buckwheat and lentil flours served as the principal flours. Buckwheat is a gluten-free pseudocereal with a high lysine content and dietary fibre [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Lentil flour is rich in high-quality plant-based protein, minerals, and prebiotic carbohydrates [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Unlike earlier formulations that use lentil flour in minimal proportions, this study successfully incorporated it at a 50% level, resulting in a protein content of 12.82 g/100 g substantially higher than in conventional gluten-free breads [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe absence of animal-derived ingredients such as eggs or milk powder renders the product suitable for vegans and individuals with lactose intolerance. Furthermore, the inclusion of almonds and hazelnuts, both rich in oleic acid and α-tocopherol, contributed to the product\u0026rsquo;s healthy fat profile and antioxidant content. These nuts are known for enhancing cardiovascular health and providing vitamin E [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], and their presence may also positively influence product shelf life due to antioxidant activity [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. The total vitamin E content (0.52 mg/100 g) aligns with values expected from nut-enriched formulations [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAlthough the fat content (11.77 g/100 g) is relatively high, the predominant fatty acid is oleic acid, which supports heart health [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The omega-3 content, at 58.85 mg/100 g, is notable, especially when compared with traditional gluten-free products [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] and contributes to cardiovascular and neurological health [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. This quantity meets approximately 11.7% of the daily recommended intake for healthy adults [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Moreover, the product contains 2,459 mg of omega-6 fatty acids per 100 g, contributing appropriately to the recommended intake range of 11\u0026ndash;22 g/day [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. However, the omega-6/omega-3 ratio was calculated as 41.8:1, which exceeds the ideal range of 4:1 to 5:1 [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e] likely due to the absence of omega-3-rich sources such as chia or flaxseed. Future research should consider reformulating to achieve a more favourable fatty acid ratio.\u003c/p\u003e\u003cp\u003eThe carbohydrate content (31.64 g/100 g) is lower than that of many commercial gluten-free breads [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Since 70% of the product\u0026rsquo;s caloric value is derived from carbohydrates, it presents a balanced glycaemic profile. The inclusion of legume flour has been shown to mitigate postprandial glycaemic response and enhance satiety [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] suggesting potential benefits for individuals with metabolic disorders.\u003c/p\u003e\u003cp\u003eThe fibre content (8.07 g/100 g) exceeds that of most gluten-free commercial products, which typically range from 3\u0026ndash;6% [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e], fulfilling approximately 25% of the recommended daily fibre intake. Dietary fibre contributes to intestinal health, glycaemic regulation, cholesterol reduction, and prolonged satiety, and also improves the shelf life of baked products through structural stabilization.\u003c/p\u003e\u003cp\u003eMoisture and ash contents were determined to be 33.36% and 2.34 g/100 g, respectively values consistent with gluten-free bread literature [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. The product maintained optimal chewability without requiring texture-modifying additives, indicating a naturally stable consistency.\u003c/p\u003e\u003cp\u003eSensory analysis confirmed high consumer acceptance for all evaluated parameters, including appearance, aroma, flavour, and mouthfeel. The contribution of nuts to chewability and the enhancement of flavour through coriander and fennel were particularly notable. Similar findings have been reported by [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e], who observed increased sensory scores in gluten-free breads containing spices. Likewise, studies by Tun\u0026ccedil; ve Kahyaoglu (2016) and Oliete et al. (2008) demonstrated improved texture and consumer acceptability with the inclusion of nut pastes [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eOverall, a gluten-free bread with a balanced nutritional profile and high sensory acceptance was successfully developed without the use of chemical additives, thickeners, or preservatives. This formulation is not only suitable for individuals with coeliac disease but also for those following plant-based or low glycaemic index diets. Furthermore, the product\u0026rsquo;s concentrated nutrient content and shelf-stability potential make it an appropriate option for crisis and emergency food supply contexts.\u003c/p\u003e\u003cp\u003eThe use of the Box\u0026ndash;Behnken design allowed efficient exploration of the formulation space with a relatively small sample size, enabling time- and cost-effective optimization. However, the study has several limitations. Firstly, the glycaemic index was estimated theoretically rather than clinically determined. Secondly, comprehensive evaluations such as shelf life, microbial safety, textural properties, and physicochemical stability were not conducted. Thirdly, the sensory panel included only a limited number of healthy individuals, without feedback from coeliac patients or those routinely following a gluten-free diet. Additionally, consumer surveys and market preference data were not collected.\u003c/p\u003e\u003cp\u003eFuture research should address these limitations by conducting \u003cem\u003ein vivo\u003c/em\u003e glycaemic assessments, stability analyses, and consumer-based preference studies across diverse demographics. Tailored product development targeting specific populations, such as children, older adults, or athletes, would further enhance the market relevance of gluten-free functional breads.\u003c/p\u003e"},{"header":"4. Conclusion","content":"\u003cp\u003eThis study successfully developed a nutritionally enriched, gluten-free functional bread using high-protein red lentil flour as the primary ingredient, optimized through a Box\u0026ndash;Behnken experimental design. The resulting formulation exhibited superior sensory properties and a well-balanced nutritional profile, characterized by elevated levels of protein, dietary fibre, and beneficial unsaturated fatty acids, particularly oleic acid. The product also demonstrated compliance with international gluten-free standards, rendering it suitable for individuals with coeliac disease, as well as for those adhering to plant-based or health-conscious diets.\u003c/p\u003e\u003cp\u003eThe compositional analysis revealed that the optimized formulation addressed several common nutritional deficiencies observed in gluten-free diets, particularly in terms of protein, fibre, and essential fatty acid intake. The successful incorporation of 50% lentil flour, a higher proportion than typically employed, contributed significantly to the product\u0026rsquo;s protein density without compromising sensory acceptability. Additionally, the inclusion of almonds, hazelnuts, and spices enhanced the organoleptic and antioxidant characteristics of the bread, contributing to its favourable reception in sensory evaluations.\u003c/p\u003e\u003cp\u003eImportantly, this study demonstrated the feasibility of producing additive-free, plant-based gluten-free bread with functional and nutritional advantages using sustainable and regionally available raw materials. The use of response surface methodology facilitated a robust and efficient optimization process, enabling the identification of an optimal ingredient matrix within a limited number of trials.\u003c/p\u003e\u003cp\u003eDespite these promising outcomes, the study presents limitations that warrant further investigation. Clinical validation of the glycaemic index, shelf-life assessment, microbial safety analysis, and broader sensory evaluations, particularly among coeliac patients and habitual gluten-free consumers, are necessary to establish the product\u0026rsquo;s market readiness and health claims. Furthermore, future studies should explore strategies to improve the omega-6/omega-3 ratio through the inclusion of additional omega-3-rich ingredients, as well as expand the application of this formulation to diverse population groups, including pediatric, geriatric, and athletic cohorts.\u003c/p\u003e\u003cp\u003eThe findings underscore the potential of lentil-based formulations as a viable alternative in the gluten-free product market, offering both nutritional and sensory benefits. This research contributes to the advancement of functional food innovation by addressing the dual imperatives of dietary health and sustainable food development.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\n\u003cp\u003eB.D.M. conceived and designed the study, developed the research methodology, and carried out the formal analysis. She led the investigation, coordinated resources, curated and managed the data, prepared the original draft of the manuscript, created the visualizations, and provided overall supervision of the research process.G.P., P.\u0026Ccedil;., and A.A. were responsible for the preparation of bread samples and the execution of sensory evaluations. They contributed to the acquisition of experimental data and participated in the interpretation of the results.M.S. contributed substantially to the writing, critical review, and editing of the manuscript, ensuring clarity, coherence, and alignment with journal requirements.\u003c/p\u003e\n\u003ch2\u003eAcknowledgement\u003c/h2\u003e\n\u003cp\u003eAcknowledgementThis study was supported by the T\u0026Uuml;BİTAK 2209-A programme. We would like to thank the Scientific and Technological Research Council of Turkey (T\u0026Uuml;BİTAK). Our sincere thanks also go to all the panellists who contributed to the product development and analysis proces-ses, as well as to the academic staff of the Departments of Nutrition and Dietetics and Gastro-nomy and Culinary Arts at Avrasya University. The authors wish to express their sincere gratitude to Assistant Professor Abdurrahman Atalay and Sefa Murat Dokuz for their valuable assistance in the English language editing of this manuscript.A preliminary version of this study was presented as an oral presentation at the Sustainable Nutrition Days: Shaping the Future with Turkish Cuisine symposium (S\u0026uuml;rd\u0026uuml;r\u0026uuml;lebilir Beslenme G\u0026uuml;nleri: T\u0026uuml;rk Mutfağı ile Geleceğe Y\u0026ouml;n Veriyoruz Sempozyumu-S\u0026Uuml;RBES2025) held in Turkey. The presentation was not published in any proceedings or academic journals.\u003c/p\u003e\n\u003ch2\u003eData Availability\u003c/h2\u003e\n\u003cp\u003eAll data supporting the findings of this study are included within the main text and its Supplementary Information. Detailed descriptions of the experimental design and the complete analysis results are provided in Supplementary File 1.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eN. R. Choudhury, \u0026ldquo;Healthy Food Market\u0026ndash;Trends, Demand \u0026amp; Consumer Shifts.\u0026rdquo; [\u0026Ccedil;evrimi\u0026ccedil;i]. Erişim adresi: https://www.futuremarketinsights.com/reports/healthy-food-market\u003c/li\u003e\n\u003cli\u003eF. Boukid, M. Mefleh, H. Mameri, ve C. M. Rosell, \u0026ldquo;Plant protein innovations in snacks and bakery: Synergy of market trends and scientific advances\u0026rdquo;, \u003cem\u003eFood Biosci.\u003c/em\u003e, c. 62, s. 105580, Ara. 2024, doi: 10.1016/j.fbio.2024.105580.\u003c/li\u003e\n\u003cli\u003eB. Boruah ve S. Ray, \u0026ldquo;Current progress in the valorization of food industrial by-products for the development of functional food products\u0026rdquo;, \u003cem\u003eFood Sci. Appl. 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Int.\u003c/em\u003e, c. 14, sy 3, ss. 259-269, Haz. 2008, doi: 10.1177/1082013208095691.\u003cbr\u003e \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Functional food, gluten-free bread, lentil flour, Box-Behnken design","lastPublishedDoi":"10.21203/rs.3.rs-7271202/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7271202/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe increasing demand for nutritionally enhanced and gluten-free functional foods has catalyzed innovation in bread formulations targeting individuals with coeliac disease and health-conscious consumers. This study aimed to develop a gluten-free bread utilizing high-protein red lentil flour, optimized through a Box\u0026ndash;Behnken experimental design. Lentil flour, buckwheat flour, and olive oil were selected as independent variables, while almonds, hazelnuts, pumpkin seeds, and spices were included at fixed levels to enhance nutritional and sensory properties. Fifteen formulations were evaluated through sensory analysis, and statistical optimization identified the ideal composition as 50 g lentil flour, 45 g buckwheat flour, and 6.5 g olive oil. The optimized bread exhibited superior sensory acceptability and a favorable nutritional profile, delivering 12.82 g protein, 8.07 g dietary fibre, 58.85 mg omega-3 fatty acids, and 0.52 mg vitamin E per 100 g, with gluten content well below the Codex Alimentarius threshold (\u0026lt;\u0026thinsp;20 ppm). The product was free from animal-derived ingredients and chemical additives, making it suitable for vegan and lactose-intolerant populations. Despite a relatively high fat content (11.77 g/100 g), the predominant fatty acid was oleic acid (64%), supporting cardiovascular health. These findings demonstrate the feasibility of producing a nutritionally dense, sensory-acceptable, and additive-free gluten-free bread using sustainable plant-based ingredients. By providing a scientifically optimized formulation that addresses key nutritional and sensory challenges in gluten-free baking, this study makes a meaningful contribution to the literature on functional food innovation, sustainable ingredient utilization, and inclusive dietary product development.\u003c/p\u003e","manuscriptTitle":"Evaluation of High-Protein Red Lentils in the Development of Gluten-Free Functional Bread","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-11 07:46:32","doi":"10.21203/rs.3.rs-7271202/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"7c5b313e-967d-42a8-8cf4-2e2343a771e5","owner":[],"postedDate":"August 11th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-08-11T07:46:34+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-11 07:46:32","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7271202","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7271202","identity":"rs-7271202","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}