European Journal of Nutrition & Food Safety 13(3): 74-82, 2021; Article no.EJNFS.67609 ISSN: 2347-5641 Effect of Natural Fermentation Period on Nutritional, Anti Nutritional, I Total Phenols, Flavonoids and Antioxidant Contents of Finger Millet Flour V. F. Abioye1 J. A. Adejuyitan1*, A. O. Adeoye1 and I. O. Gbadegesin1 1 Department of Food Science, Faculty of Food and Consumer Sciences, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria. Authors’ contributions This work was carried out in collaboration among all authors. Authors VFA and JAA designed the study, performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript. Authors AOA and IOG managed the analysis of the study. All authors read and approved the final manuscript. Article Information DOI: 10.9734/EJNFS/2021/v13i330392 Editor(s): (1) Dr. Rasha Mousa Ahmed Mousa, University of Jeddah, Jeddah, Saudi Arabia. Reviewers: (1) Borşa Andrei, University of Agricultural Sciences and Veterinary Medicine, Romania. (2) Dele Raheem, University of Lapland,Finland. Complete Peer review History: http://www.sdiarticle4.com/review-history/67609 Original research Article Received 02 March 2021 Accepted 06 May 2021 Published 24 June 2021 ABSTRACT Aims: This study determined the changes in the chemical and nutritional composition of naturally fermented finger millet studied at ambient temperature (28±2°C) for 72 h. Study Designs: Finger millet seeds were cleaned and fermented (72 h; 28±2°C). Samples were taken at 24 h interval and dried at 50°C for 48 h. Methodology: The fermented finger millet samples were analyzed for microbial, biochemical changes, chemical, proximate and mineral composition. Results: Biochemical changes showed a drop in pH from 6.74 to 6.04 while titratable acidity (lactic acid equivalent) increased from 0.04 to 0.62% after 72 h. The moisture, protein, ash, fat, fibre and carbohydrate were in the ranges of 7.08-9.449%; 5.31-7.274%; 1.10-3.392%, 1.296-2.47%, 1.1542.46% and 77.44-81.58%, respectively. Significant increase were observed in the mineral composition with phosphorus, potassium, calcium, sodium, and iron identified in the fermented finger millet flour in the ranges of 93.5-176 mg/100 g; 171-247.5 mg/100 g; 87.04-196.5 mg/100 g; 1.30-3.075 mg/100 g and 5.28-11.95 mg/100 g, respectively. Tannin, oxalate, phytate and trypsin _____________________________________________________________________________________________________ *Corresponding author: Email: jadejuyitan@lautech.edu.ng; Abioye et al.; EJNFS, 13(3): 74-82, 2021; Article no.EJNFS.67609 were in the ranges of 1.537 to 3.23 mg/100 g; 0.875 to 1.59 mg/100 g; 0.195 to 0.85 mg/100 g. and 2.731 to 6.23 mg/100 g, respectively. The total phenols and total flavonoids ranged between 11.605-40.29 mg/ 100 g and 63.36 -172.872 mg/100g while the 2,2-diphenyl-1-picrylhydrazyl (DPPH) of the flour samples ranged between 28.109 and 68.238 mg/ml. Microorganisms identified were Bacillus cereus, Lactobacillus Plantarum, Lactobacillus casei and Lactobacillus brevis. This study shows that fermentation decreased the anti-nutrients, increased the proximate and minerals contents and also improved the anti-oxidative properties of finger millet flour. Keywords: Finger millet; fermentation; anti-nutrients; antioxidants. 1. INTRODUCTION used in processing of cereals such as sorghum and millets. During fermentation, flour undergoes major biochemical reactions such as starch hydrolysis, sugar transformation and softening which have been reported to improve nutritional quality of cereal grain, reduce the anti-nutrients and increase bioavailability of micro nutrients [21,22]. Finger millet (Eleusine coracana) commonly known as ragi and mandua in India is one of the minor cereals that is grown extensively in various regions of India and Africa [1,2]. The grains contain about 5–8% protein, 1–2% ether extractives, 65–75% carbohydrates, 15–20% dietary fiber and 2.5–3.5% minerals [3,4]. Finger millet is considered as one of the most nutritious cereals with highest content of calcium (344 mg/100g) and potassium (408 mg/100g) [5,6]. It has nutraceutical properties and it is also recognized for its health beneficial effects, such as antidiabetic, antitumorigenic, antidiarrheal, anti inflammatory, atherosclerogenic effects, antioxidant and antimicrobial properties [2]. Finger millet is referred to as poor man’s food due to its ability to be stored safely for many years without infestation by insects and pests [7,8,9]. The microbial population and biochemical changes during the fermentation of finger millet have been reported [23]. Effects of fermentation of finger millet on the primary nutrients have also been reported [24]. However, there is scanty information on the effects of fermentation on the nutrients, anti-nutrients and antioxidative properties of finger millet. This study, therefore, determined the effect of natural fermentation on the biochemical changes, proximate, minerals, anti-nutrients, phenolic, flavonoids and antioxidant properties of finger millet. 2. MATERIALS Finger millet is an important cereal due to its excellent storage properties and the nutritive value, which is higher than that of rice and equal to that of wheat [10,8]. It is a good source of micronutrients (calcium, iron, phosphorus, zinc and potassium) and essential amino acids (valine, methionine, isoleucine, threonine and tryptophan) which are essential for human body [11,12,13]. It is gluten-free, has low Glycemic Index and. Millet is used as whole flour mostly for traditional food preparation and can be consumed after processing in form of noodles, biscuit, muffins, vermicelli, pasta and bread [14,15,16]. Finger millet (Eleusine coracana) was obtained from Jos, Plateau State and identified at Ladoke Akintola University of Technology (LAUTECH), Ogbomoso, Research Farm Unit. 2.1 Production of Fermented Finger Millet Flour About 2500 grams of grains were sorted out by removing the debris, stones and adhering substances. The cleaned seeds were shared into five different portions of 500 g each. Each of the portion was steeped in water (1:5 w/v) and allowed to ferment at 28±2 °C for maximum of 72 h. The fermented grains were harvested after 24 h, washed, dried at 50 °C and milled into fine flour (300 µm). The flours were packaged in zip lock bags until further uses Bioavailability of some of these nutrients in finger millet is limited by the presence of anti-nutrients such as tannins, phytates, and oxalates [17,12,18]. Processing methods such as boiling, soaking, roasting, germination and fermentation have been reported to reduce the anti-nutrient content of finger millet and thus enhance the biological availability of the nutrients [19,20]. Fermentation is one of the oldest methods widely 2.2 Analyses Lactic acid bacteria were enumerated and identified using the method described by [25]. 75 Abioye et al.; EJNFS, 13(3): 74-82, 2021; Article no.EJNFS.67609 3.2 Proximate Millet Ten grams of each of the fermented sample was homogenized with 90 ml of 0.85% (w/v) saline and serially diluted. One hundred micro liter of the sample suspension was spread on MRS agar media after plating, the sample plates were incubated anaerobically at 37°C for 48 h. Colonies were selected and purified by restreaking to obtain purified strains. The pH of each of the fermented sample was measured using a pH meter (Jenway Model). The titratable acidity (lactic acid, %) was determined by measuring the amount of 0.1 N NaOH necessary to adjust to pH 8.3. Microbial isolates were carried out as described by [23]. The fermented flour samples were analyzed for moisture, ash, crude fibre, protein (N*6.25), crude fat and the carbohydrate determined by difference according to the method described by [26]. The minerals present in the fermented flour samples were quantified using the dry-ash techniques according to (AOAC, 2010). The tannin content of the flour samples was determined using FolinCiocalteu method as described by [27]. Phytate content was determined using rapid colourimetry method as described by [28]. The oxalate content of the samples was determined using titration method as described by [29]. Total Phenolic Content (TPC) of the extracts was determined using spectrophotometry as described by [30]. Aluminium chloride colourimetric method was used for flavonoids determination [31]. The DPPH scavenging activity was carried out by the method of [32]. The results of the experiment were subjected to analysis of variance (ANOVA) and the means separated with the use of Duncan’s multiple range test. Composition of Finger The proximate composition of the fermented flour samples is as shown in Table 1. The moisture content ranged from 7.08% to 9.449% with increase in moisture content as the fermentation hours increased. There was increase in the moisture content with increase in fermentation time. The protein content of the fermented finger millet also followed the same trend as the hour of fermentation rose up. This is in line with previous reports of other researchers, [33] reported a rise in protein content of finger millet as the fermentation hour rose up. Similar reports were recorded for pearl millet by other researchers [37,38,39,40,22] while some others reported increase in some cereals during fermentation [41,42,43] reported improvement in amino acid balance as well as the sensory quality and nutritional value of the cereal grains during fermentation. Increase in protein content could be attributed to the loss of dry matter, mainly carbohydrates or due to the action of extracellular enzymes produced by the fermenting microorganisms [44]. The increase may also be partly as a result of degradation of complex protein by microorganism thereby releasing peptides and amino acids [45]. The ash content of the fermented finger millet flour ranged from 1.10 9% to 3.392% with gradual increase with increase in fermentation time. The result is in line with other reports that minerals increased with fermentation period [39,45]. Minerals from plant sources have very low bioavailability because they are found complexed with non-digestible material such as cell wall polysaccharides [46,47]. Fermentation was also reported to increase bioavailability of calcium, phosphorous, and iron likely due to degradation of oxalates and phytates that complex with minerals thereby reducing their bioavailability [48]. 3. RESULTS AND DISCUSSION 3.1 Changes in pH and Titratable Acidity of Finger Millet and Microbial Isolates There was a decrease in the lipid content of the finger millet flour from 9.47 to 7.296. This is in conformity with the findings of [24] who reported about 42.9% reduction in the total fat content in fermented finger millet, [40] also reported a decrease in fat content of pearl millet during fermentation. The low lipid content observed in the fermented sample could help in improving the shelf life of fermented finger millet flour by decreasing the chances of rancidity and it may also contribute to the low energy value of the samples. The carbohydrate content of the fermented finger millet flour ranged between The prominent microorganisms identified were Bacillus cereus, Lactobacillus Plantarum, Lactobacillus casei and Lactobacillus brevis. The pH decreased from 7 to 6.04 with increase in the hour of fermentation while titratable acidity increased from 0.04 to 0.3 with increase in the hour of fermentation as shown in Fig 1. This report is in line with the report of other researchers [33,34,35, and 36] that the pH decreased while titratable acidity increased with as fermentation period increased. 76 Abioye et al.; EJNFS, 13(3): 74-82, 2021; Article no.EJNFS.67609 3.3 Mineral Composition of Finger Millet Flour 71.435% and 74.59%%. There was gradual decrease in the carbohydrates with increase in fermentation time. This is in line with reports of other researchers who had worked on other millets such as [49] and [21] who worked on pearl millet and [24] who reported on foxtail millet. This observation is in conformity with previous reports that carbohydrates are a major carbon source for fermenting microbes [50]. The fiber content values of the sampled finger millet flour were between 0.893% and 2.154%. The mineral contents of the fermented finger millet are presented in Table 2. The results indicated increase in the mineral contents with increase in the fermentation time. This is in line with other researchers that fermentation increases the bioavailability of minerals in cereals [51]. Chart Title pH and Titable acidity 8 7 6 5 4 3 2 1 0 0h 24 h 48 h 72 h fermentation period in h Ph titrable acidity Fig. 1. Changes in pH and total titratable acidity during the natural fermentation of finger millet Table 1. Proximate composition of finger millet flour (%) Samples A B C D Moisture Content 7.080±0.282a 8.289±0.001b 9.121±0.001c 9.449±0.001d Crude Protein 5.310±0.014b 5.241±0.001a 6.019±0.001c 7.274±0.006d Ash Crude Lipid Crude Fibre CHO 1.100±0.00a 2.019±0.001b 2.080±0.014c 3.392±0.003d 2.470±0.014d 1.697±0.001c 1.437±0.001b 1.297±0.002a 2.460±0.014d 1.893±0.004c 1.319±0.000b 1.154±0.006a 81.580±0.028d 80.861±0.001c 80.031±0.001b 77.435±0.001a Values represent means of triplicate reading, follow by different lowercase letter Means within the same row with different alphabets are significantly different (p≤0.05) Sample A: Samples that were not fermented (Control); Sample B: Samples fermented for 24 h; Sample C: Samples fermented for 48 h;Sample D: Samples fermented for 72 h Table 2. Mineral composition of fermented finger millet flour (mg/100 g) Samples A B C D P 93.500±0.141a 185.000±1.414c 255.000±1.414d 176.000±1.000b K 171.000±1.414a 228.000±0.000b 239.000±1.414c 247.500±0.141d Ca 87.040±0.000a 108.800±0.141b 156.200±0.283c 196.500±0.141d Na 1.300±0.000a 2.649±0.000c 2.351±0.001b 3.008±0.001d Fe 5.280±0.014a 10.800±0.141b 10.700±0.141b 11.450±0.495b Values represent means of triplicate reading, follow by different lowercase letter Means within the same row with different alphabets are significantly different (p≤0.05) Sample A: Samples that were not fermented (Control);Sample B: Samples fermented for 24 h;Sample C: Samples fermented for 48 h;Sample D: Samples fermented for 72 h 77 Abioye et al.; EJNFS, 13(3): 74-82, 2021; Article no.EJNFS.67609 Table 3. Anti-nutritional factor of finger millet flour (mg/100g) Samples A B C D Oxalate 1.590±0.014d 1.325±0.001b 1.410±0.014c 0.875±0.000a Trypsin 6.230±0.014d 5.338±0.001c 3.121±0.001b 2.731±0.001a Phytate 0.850±0.014d 0.594±0.003c 0.204±0.003b 0.195±0.001a Tannin 3.230±0.014d 2.431±0.001c 1.896±0.000b 1.537±0.001a Values represent means of triplicate reading, follow by different lowercase letter Means within the same row with different alphabets are significantly different (p≤0.05) Sample A: Samples that were not fermented (Control); Sample B: Samples fermented for 24 h; Sample C: Samples fermented for 48 h; Sample D: Samples fermented for 72 h Chart Title 250 200 150 100 50 0 A B C Total Phenols mg/100 g Total Flavonoids D DPPH Fig. 2. Total phenols, total flavonoids and the antioxidant content of finger millet flour Sample A: Samples that were not fermented (Control);Sample B: Samples fermented for 24 h;Sample C: Samples fermented for 48 h;Sample D: Samples fermented for 72 h 3.4 Antinutritional composition of Finger Millet the free nutrients which improves availability of these nutrients [52,53,54]. The result of anti-nutritional contents of the fermented finger millet is as shown in Table 4. Finger millet has been reported to contain the highest amount of anti-nutrients among other millets. There was significant reduction (p<0.05) in phytate and tannin content. Phytic acid has been reported as one of the antinutritional factors common in cereals responsible for binding minerals and thus making them not readily bioavailable (52, 42). The phytate in the finger millet decreased from 0.85 mg/100 g to 0.195 mg/100 g, the result is in line with other researchers who reported decrease in phytic acid with fermentation (52; Anthony and Chandra, 1998, Osman, 2011). The fermenting microorganisms are responsible for the cleavages of tannin-protein, tannic acid-starch and tannin-iron complexes thereby releasing 3.5 Antioxidant Properties of Finger Millet Flour the The total phenols, total flavonoids and the antioxidant content in the flour samples are as shown in Fig 2. The total phenols, total flavonoids and the antioxidant content were in the ranges of 11.605-40.29 mg/ 100 g, 63.36 172.872 mg/100g and 28.109 and 68.238 mg/ml, respectively. The total phenolic contents decreased with increase in fermentation time The effects of fermentation on phenolic compounds were reported to be a factor of grain types, microorganism species and fermentation conditions (temperature, pH, and time) [55,56]. The flavonoids content increased with increase in fermentation time, this agrees with findings of [40] that fermentation enhanced the flavonoids 78 Abioye et al.; EJNFS, 13(3): 74-82, 2021; Article no.EJNFS.67609 content in fermented pearl millet food. Antioxidants are substances that neutralize the harmful free radicals in our bodies. Anti-oxidant act as free “radical scavengers” and hence prevent or slow the damage done by free radicals. Anti-oxidants are also used to prevent food quality meanly because the slow oxidative deterioration of lipids. Anti-oxidants function as reducing agents, ultimately removing free radical intermediates and preventing further oxidation [57]. 5. 6. 4. CONCLUSION This study has revealed that fermentation of finger millet can increase the protein, mineral content with significant reduction in the antinutrients thereby increasing the bioavalability of these nutrients. Fermentation also enhanced the polyphenols potentials of the crop. 7. ACKNOWLEDGEMENT The authors acknowledged the financial support granted by TETFUND in carrying out this research work 8. COMPETING INTERESTS Authors have interests exist. declared that no competing 9. REFERENCES 1. 2. 3. 4. Gull A, Jan R, Nayik GA, Prasad K, Kumar P. Significance of finger millet in nutrition, health and value added products: a review. Journal of Environmental Science, Computer Science and Engineering and Technology. 2014;3(3):1601-1608. Chandra S, Pallavi, Sharma AK. Review of finger millet (Eleusine coracana (L.) Gaertn): A power house of health benefiting nutrients. Food Science and Human Wellness. 2016;(5):149–155. Chethan S, Malleshi NG. Finger millet polyphenols: Characterization and their nutraceutical potential. America Journal Food Technology. 2007;2:582–592. Devi PB, Vijayabharathi R, Priyadarisini VB.. Health benefits of finger millet (Eleusine coracana L.) polyphenols and dietary fiber: a review. J Food Sci Technol. 2014;51(6):1021–1040. DOI: 10.1007/s13197-011-0584-9 10. 11. 12. 13. 79 Shobana S, Krishnaswamy K, Sudha V, Malleshi NG, Anjana RM, Palaniappan L, Mohan V. Finger millet (Ragi, Eleusine coracana L.): a review of its nutritional properties, processing, and plausible health benefits. Adv Food Nutr Res. 2013;69:1‐39. DOI: 10.1016/B978-0-12-410540-9.000016 Kumar A, Metwal M, Kaur S, Gupta AK, Puranik S, Singh S, Singh M, Gupta S, Babu BK, Sood S, Yadav R Nutraceutical value of finger millet [Eleusine coracana (L.) Gaertn.], and their improvement using Omics Approaches. Front. Plant Sci. 2016;7:934. DOI: 10.3389/fpls.2016.00934 Bekele AF, Getahun DW, Derejem AB. Determination of optimumratesof nitrogen and phosphorus fertilization for finger millet (Eleusine coracana L. Gaertn) production at Assosa Zone, in Benishangul – Gumuz Region of Ethiopia. Advances in Sciences and Humanities. 2016;2(1):1-6. DOI: 10.11648/j.ash.20160201.1 Gull A, Gulzar Ahmad N, Prasad K, Kumar P. Technological, processing and nutritional approach of finger millet (Eleusine coracana) - A mini review. J Food Process Technol. 2016;7:593. DOI: 10.4172/2157-7110.1000593 Aisoni JE, Yusha’u M, Orole OO.Processing effects on physicochemical and proximate composition of finger millet (Eleusine coracana). Greener Journal of Biological Sciences. 2018;8(2):014-020. Van Wyk BE, Gericke N. People’s plants. A Guide to Useful Plants of Southern Africa. Pretoria: Briza Publications. 2000;23-25. FAO. Amino acid scoring pattern. In: Protein quality evaluation, FAO/WHO Food and Nutrition Paper, Italy. 1991;12-24. Mbithi-Mwikya S, Ooghe W, Van Camp J, Nagundi D, Huyghebaert A. Amino acid profile after sprouting, Autoclaving and lactic acid fermentation of finger millet (Eluesine coracana) and kidney beans (Phaseolus vulgaris L.) J. Agric. Food Chem. 2000;48 (8):3081-3085. Thapliyal V, Singh K. Finger millet: Potential millet for food security and power house of nutrients. International Journal of Research in Agriculture and Forestry. 2015;2(2):22-33. Abioye et al.; EJNFS, 13(3): 74-82, 2021; Article no.EJNFS.67609 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. Krishnan R, Dharmaraj U, Manohar RS, Malleshi NG. Quality characteristics of biscuits prepared from finger millet seed coat‐based composite flour. Food Chem. 2011;129(2):499– 506. Gull A, Prasad K, Kumar P. Nutritional, antioxidant, microstructural and pasting properties of functional pasta. Journal of the Saudi Society of Agricultural Sciences. 2016;17(2):147-153. Rathore T, Singh R, Kamble DB, Upadhyay A, Thangalakshmi S. Review on finger millet: Processing and value addition. The Pharma Innovation Journal. 2019;8(4):283-291. Gibbs-Russell P, Deosthale YG. In vitro availability of iron and zinc in white and coloured ragi (Eleusine coracana): role of tannin and phytate. Plant Foods Human Nutrition. 1989;38:35-41. Popova A, Mihaylova D. Antinutrients in plant-based foods: A review. The Open Biotechnology Journal. 2019;13(1):68-76. Hotz C, Gibson RS.Traditional foodprocessing and preparation practices to enhance the bioavailability of micronutrients in plant-based diets. The Journal of Nutrition. 2007;137(4):1097– 1100. Available:https://doi.org/10.1093/jn/137.4. 1097Nkhata et al., 2018 Abioye VF, Ogunlakin GO, Taiwo G. Effect of germination on anti-oxidant activity, total phenols, flavonoids and anti-nutritional content of finger millet flour. J. Food Process Technol. 2018;9:719. DOI: 10.4172/2157-7110.1000719 Osman MA. Effect of traditional fermentation process on the nutrient and antinutrient contents of pearl millet during preparation of Lohoh. Journal of the Saudi Society of Agricultural Sciences. 2011;10:1–6. Gupta RK, Gangoliya SS, Singh NK. Reduction of phytic acid and enhancement of bioavailable micronutrients in food grains. J Food Sci Technol. 2015;52(2):676–684. DOI: 10.1007/s13197-013-0978-y Antony U, Chandra T. Microbial population and biochemical changes in fermenting finger millet (Eleusine coracana). World Journal of Microbiology and Biotechnology. 1997;13:533–537. Available:https://doi.org/10.1023/A:101856 1224777. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 80 Antony U, Sripriya G, Chandra TS. Effect of Fermentation on the primary nutrients in finger millet (Eleusine coracana). Journal of Agricultural and Food Chemistry. 1996;44(9):2616-2618. DOI: 10.1021/jf950787q. Sangita B, Apoorva S, Manisha M, Satish KS. Isolation and characterization of lactic acid bacteria from fermented foods. Vegetos. 2013;26(2):325-330. AOAC, Association of official Analytical Chemists. Official methods of analysis, th 18 ed. Association of Official Analytical Chemists, Washington, D. C; 2010. Hailer M, Kang WH. Antioxidant activity, total polyphenol, flavonoid and tannin contents of fermented green coffee beans with selected yeasts. Fermentation. 2019;5:29:1-13. Latta M, Eskin M. A Simple and rapid colorimetric method for phytate determination. J. Agric. Food Chem. 1980;28:313-315. Debi PM, Nivedita M, Harshal BM, Pinaki S, Sidharth M, Devi PS. Determination of seasonal and developmental variation in oxalate content of Anagallis arvensis plant by titration and spectrophotometric method. The Pharma Innovation Journal. 2017;6(6):105-111. Singleton VL, Orthofer R, LamuelaRaventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin–Ciocalteau reagent. Method. Enzymol. 1999;299:152– 178. Luximon-Ramma AT, Bahorun MA, Soobratte, Aruoma OI. Antioxidant activities of phenolic, proanthrocyanidin and flavanoid components in extracts of Cassia fistula. J. Agric. Food Chem. 2002;50:5042-5047. Blois MS. Antioxidant determination by use of stable free radicals. Nature. 1985;29:1199–1200. Antony U, Chandra T. Antinutrient reduction and enhancement in protein, starch, and mineral availability in fermented flour of finger millet (Eleusine coracana). Journal of Agricultural and Food Chemistry. 1998;46(7):2578-2582. DOI: 10.1021/jf9706639 Arora S, Jood S, Khetarpaul N, Goyal R. Effect of germination and fermentation on pH. Titratable acidity and chemical composition of pearl millet based food Abioye et al.; EJNFS, 13(3): 74-82, 2021; Article no.EJNFS.67609 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. blends. Acta Alimentaria. 2009;38(1):107115. Jood S, Kheterpaul N, Goyal R. Effect of germination and probiotic fermentation on pH, titratable acidity, dietary fibre, β-glucan and vitamin content of sorghum based food mixtures. Journal of Nutrition and Food Sciences. 2012;2:9. DOI: 10.4172/2155-9600.1000164 Ilango S, Antony U. Assessment of the microbiological quality of koozh, a fermented millet beverage Shankar. 2014;8(3):308-312. Ali AM, El-Tinay HA, Abdalla HA. Effect of fermentation on the in vitro digestibility of pear millet. Food Chem. 2003;80:51–54. Hassan AB, Mohamed Ahmed IA, Osman NM, Eltayeb MM, Osman GA, Babiker EE. Effect of processing treatment followed by fermentation on protein content and digestibility of pearl millet (Pennisetum typhoideum) cultivars. Pakistan J. Nutr. 2006;5(1):86–8. Inyang CU, Zakari UM. Effect of germination and fermentation of pearl millet on proximate, chemical and sensory properties of instant, Fura–Nigerian cereal food; 2008. Gupta V, Nagar R. Effect of cooking, fermentation, dehulling and utensils on antioxidants present in pearl millet rabadi a traditional fermented food. J Food Sci Technol. 2010;47(1):73‐76. DOI: 10.1007/s13197-010-0018-0 Day CN, Morawicki RO. Effects of fermentation by yeast and amylolytic lactic acid bacteria on grain sorghum protein content and digestibility. Hindawi Journal of Food Quality. 2018;1–8. DOI: 10.1155/2018/3964392 Nkhata SG, Ayua E, Jean‐Bosco S. Fermentation and germination improve nutritional value of cereals and legumes through activation of endogenous enzymes. Food Sci Nutr. 2018;6:2446– 2458. Ramashia SE, Tonna AA, Eastonce TG, Stephen MT, Afam IOJ. Processing, nutritional composition and health benefits of finger millet in sub-saharan Africa. Food Sci. Technol. 2019;39(2). Available:https://doi.org/10.1590/fst.25017 Hamad AM, Fields ML. Evaluation of the protein quality and available lysine of germinated and fermented cereal. Journal of Food Science. 1979;44:456–459. DOI: 10.1111/j.1365-2621.1979.tb03811.x 45. 46. 47. 48. 49. 50. 51. 52. 53. 54. 81 Pranoto Y, Anggrahini S, Efendi Z. Effect of natural and Lactobacillus plantarum fermentation on invitro protein and starch digestibilities of sorghum flours. Food Bioscience. 2013;2:46–52. DOI: 10.1016/j.fbio.2013.04.001 Torre M, Rodriquez R, Saura‐Calixto F. Effects of dietary fiber and phytic acid on mineral availability. Critical Reviews in Food Science and Nutrition. 1991;30:1–22. DOI: 10.1080/10408399109527539 Claire H, Peter R, Cathrina HE, Myriam ML, Gundy. Plant cell walls: Impact on nutrient bioaccessibility and digestibility. Advanced Research in Food digestion. 2020;9(2)201. Sripriya G, Antony U, Chandra TS. Changes in carbohydrate, free amino acids phytate and HCL extractability of minerals during germination and fermentation of finger millet (Eleusine coracana). Food Chemistry. 1997;58:345–350. DOI: 10.1016/S0308-8146(96)00206Khetarpaul N, Chauhan BM. Effect of germination and fermentation on In vitro starch and protein digestibility of pearl millet. Journal of Food Science. 1990;55:883–884. DOI: 10.1111/j.1365-2621.tb05261.x Adams MR. Topical aspects of fermented foods. Trends Food Sci Tech. 1990;1:141– 144. DOI: 10.1016/0924-2244(90)90111-B Makokha AO, Oniango RK, Njoroge SM, Kamar OK. Effect of traditional fermentation and malting on phytic acid and mineral availability from sorghum (Sorghum bicolor) and funger millet (Eleusine caracana) grain varieties grown in Kenya. Food Nutr Bull. 2002;23:241–245. Liang J, Han BZ, Nout MJR, Hamer RJ. Effects of soaking, germination and fermentation on phytic acid, total and in vitro soluble zinc in brown rice. Food Chemistry. 2008;110:821–828. DOI: 10.1016/j.foodchem.2008.02.064 Sharma A, Kapoor AC. Levels of antinutritional factors in pearl millet as affected by processing treatments and various types of fermentation. Plant Food Hum Nutr. 1996;49;241–252. Available:https://doi.org/10.1007/BF01093 221 Onweluzo JC, Nwabugwu CC. Fermentation of millet (Pennisetum americanum) and pigeon pea Abioye et al.; EJNFS, 13(3): 74-82, 2021; Article no.EJNFS.67609 55. (Cajanuscajan) seeds for flour production: Effects on composition and selected functional properties. Pakistan Journal of Nutrition. 2009;8(6):737-744. Onuoha EC, Orukotan AA, Ameh JB. Effect of fermentation (natural and starter) on the physicochemical, anti-nutritional and proximate composition of pearl millet used for flour production, American Journal of Bioscience and Bioengineering. 2017;5(1)12-16. 56. 57. DOI: 10.11648/j.bio.20170501.13 Oseni K. A review on the status of the phenolic compounds and antioxidant capacity of the flour: Effects of cereal processing, International Journal of Food Properties. 2017;20:sup1:S798-S809. DOI: 10.1080/10942912.2017.1315130 Rice-Evans CA, Miller NJ, Bolwell. The relative antioxidant activities of plant derived phenolic flavonoid. Free Radic. Res. 1995;22:375-383. © 2021 Abioye et al.; This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Peer-review history: The peer review history for this paper can be accessed here: http://www.sdiarticle4.com/review-history/67609 82