Food Chemistry Advances 4 (2024) 100633

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Food Chemistry Advances

journal homepage: www.elsevier.com/locate/focha

Gluten free sourdough breads from pearl millet-Bambara nut and pearl
millet-soybean paste: Evaluation of the proximate, functional properties of
the flour blends and the bread nutritional indexes
I.N. Okwunodulu a, *, G.N. Egbuta a, F.U. Okwunodulu b, C.M. Ojinnaka a, S.C. Onyeiwu c
a
Department of Food Science and Technology, Michael Okpara University of Agriculture, Umudike PMB 7267 Abia State, Nigeria
b
Chemistry Department, Michael Okpara University of Agriculture, Umudike PMB 7267 Abia State, Nigeria
c
Department of Food Tecnology, Kaduna Polytechnology, Kaduna State, Nigeria

A R T I C L E I N F O A B S T R A C T

Keywords: Wheat bread (WB) triggers celiac disease to the vulnerable consumers which has no cure except controlled or
Non-gluten cereals abstinence from gluten injection in the diet. Besides, wheat importation is costly which reflects on bread cost.
Sourdough breads This study aimed at elaborating and analyzing gluten free sourdough bread (SB) from pearl millet-Bambara nut
Micronutrients
and pearl millet-soybean composite flours and compared with wheat flour bread. The results revealed variations
Physical properties
Acceptability
(p < 0.05) in moisture, dry matter, ash, crude protein, crude fibre, fat, carbohydrate and energy contents of SBs
between 33.37–33.47 %, 66.54–66.63 %, 1.93–2.15 %, 9.84–10.36 %, 1.08–1.17 %, 3.78–6.35 %, 46.82–49.69
% and 276.38–294.13 Kcal respectively. While that of WB were 34.71, 63.29, 2.10, 50.51, 0.84, 4.87 and 46.95
% respectively. Micronutrient composition varied (p < 0.05) from 49.42 to 51.77, 63.64–68.50, 0.78–1.06,
0.26–0.76, 0.19–0.23 and 8.27–9.53 mg/100 g respectively for calcium, phosphorous, iron, vitamins B1, B2 and
C. Respective WB values were 42.85, 59.17, 1.17, o.23, 0.17 and 7.62 mg/100 g. Physical properties varied (p <
0.05) from 0.35 to 0.95 cm, 172.13–219 cm3, 214–219 g and 8.06–8.50 cm3/g and that of WB were 4.50 cm,
196.70 cm3 237 g and 8.30 cm3/g respectively, for oven spring, loaf volume, loaf weight and specific loaf
volume. Most of the chemicaland physical parameters have significant (p < 0.01) correlation. Acceptability
scores of the SBs varied (p > 0.05) from 5.20 to 5.85 while that of WB was 7.550. Millet-soybean paste (50: 50)
produced a better SB with higher nutrient content and acceptability score than that from millet-Bambara nut
composite flour. Varying their blending ratios could give better results.

Introduction induced enteropathy like chronic intestinal disorder which symptoms

include diarrhea, abdominal pains and fatigue (Aguirre et al., 2023).
Bread is one of the most commonly produced and consumed baked Millets are small seeded cereals grown around the world which had
product globally for ages as an affordable staple food either leavened adapted well to Africa’s semi-arid conditions unlike wheat and rice.
with yeast or sourdough bread (Goesaert et al., 2005). Wheat flour is Among the nine species cultivated globally, pearl millet is the most
most commonly used for bread production due to the presence of gluten, widely grown in Africa which produces the largest seeds and is the most
but can be replaced to a certain degree with other grains such as rye, commonly consumed specie. Millet is a good source of micronutrients
soybean flour among others. Wheat protein is low in essential amino and energy which protein is very close to that of wheat (USDA, 2012).
acids like lysine, methionine and threonine content which are desired by The protein is rich in essential amino acids like methionine but lacks
diabetics and gastro-intestinal patients. Wheat consumption by some lysine and threonine. Millets are called “nutri-cereals” because of their
people triggers allergies, intolerance and celiac disease (Shewry et al., abundant micronutrient and phytochemicals that lowers cholesterol and
2002) which unfortunately has no cure but can be controlled by cancer risk (Coulibaly et al., 2011). Millets lacks gluten but contains
avoiding gluten intake. Celiac disease is a myriad of health conditions some anti-nutrients like oxalate, phytic acid, tannins and polyphenols
derived from the body immune response to gluten. It is a common food (Chikwendu et al., 2014). The grits and flours make common composite

* Corresponding author.
E-mail address: [email protected] (I.N. Okwunodulu).

https://doi.org/10.1016/j.focha.2024.100633
Received 17 October 2022; Received in revised form 27 January 2024; Accepted 28 January 2024
Available online 5 February 2024
2772-753X/© 2024 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
I.N. Okwunodulu et al. Food Chemistry Advances 4 (2024) 100633

flours with other cereals for production of cookies, cakes and bread from matrix. These amino acids not only enrich sourdoughs nutritionally but
non-gluten sources (Hama et al., 2011). also improve flavour, rheology aroma and taste development of the
Soybean is an excellent source of cheap protein, lipid and niacin bread (Gänzle & Gobbetti, 2013). Acetic acid oxidases ethanol and lactic
which made it one of the richest valued plant foods consumed globally. acid produced by LAB to increase greatly the bread flavour The acidic
They are rich in micro and macro nutrients such as sodium, zinc, copper, medium preserves the dough (Bengar et al., 2022), acts as natural dough
potassium, iron, phosphorous, magnesium and manganese. Soybean conditioner, improves flavour and gas retention resulting in good
contains health promoting phytochemicals with relatively low antioxi­ product texture, massive bread volume and aroma (Axel et al., 2017)..
dant properties and anti-nutritional factors (Shi et al., 2010). Also, acid medium activates phytases, proteases and pentosanases
Bambara nut is a legume of African origin with balanced essential thereby contribute to dough biochemical changes. Bioactive compounds
amino acids. It contains high carbohydrates, adequate proteins, dietary are also produced during fermentation. Besides, during fermentation,
fibre, good source of vitamins and minerals, but low in fat (Alhassan fatty acids of the flour are catalysed by lipoxygenase and
et al., 2015). The flour can be used for the production of cakes and hydroperoxide-lyase to produce volatile aldehyde that contributes to
biscuit fortification. bread flavour. Also, oligosaccharide produced improved the shelf life of
Sourdough technology (ST) is the oldest biotechnological processes baked goods by lowering the evaporation rate of the bound water
employed for fermenting cereal flours for the manufacturing of baked resulting in reduced staling (Petel et al., 2017).
products (Gobbetti et al., 2019). Sourdough is spontaneously (natural) During baking of the fermented sourdough, as the temperature in­
fermented dough (Chavan & Chavan, 2011) produced from mixture of creases, it causes the yeast to produce carbon dioxide and alcohol that
gluten free cereal flours, water, consortium of mainly wild hetero­ cause suden volume rise (oven spring) and stops at 60 ◦ C while amylose
fermentative LAB (Lactobacillaceae family) as leavening agent., wild activity increase on starch stops at 95 ◦ C after starch breakdown by
yeasts (Gobbetti et al., 2019) and acetic acid bacteria (AAB) (Comasio chemical reactions into dextrin sugar (dextrinization). The flour protein
et al., 2020) for production of gluten-free baked products. The cereals is broken easily as the moisture is baked off to release amino acids that
include rice, corn, or sorghum (Smidova & Rysova, 2022), millet combine with sugar resulting in Maillard browning and distinct flavor
buckwheat, quinoa, amaranth, canihua and natural gluten-free foods between 140 and 165 ◦ C. Finally, at about 170 ◦ C, the sugar molecules
like potatoes, tapioca, nuts, oilseeds, legumes, fruits and vegetables polymerizes to form coloured caramels that contribute to sweet flavor
(Gabrovská et al., 2015). Their flours and their ingredients, physiolog­ and colour (Enest, 2017). Sourdough products are associated with
ical properties of the flour-water mixture (temperature, pH, titratable longer shelf life, low glycemic index, higher sensorial quality and better
acidity, oxygen tension and fermentation time) are the sources of an nutritional value due to reduced anti-nutrients (Oboh et al., 2003;
additional microbial inoculum and co-substrates for back slopping Gobbetti et al., 2019) compared with other types of bread. This study
sourdough production (Minervini et al., 2018; Oshiro et al., 2020). aimed at producing, evaluating and comparing gluten-free breads from
After production, the sourdough is further subjected to a back slop­ millet-Bambara nut and millet-soybean paste composite sourdoughs.
ping fermentation process for traditional sourdoughs or a single step
fermentation (hours to several days) for industrial sourdoughs (De Vuyst Materials and methods
et al., 2017). For bakery practice, sourdough is produced by back slop­
ping using sourdough starter cultures or ingredients that bypass the Pearl millet, soybean Bambara nut and all the ingredients used in
typical microbiota succession involved in back slopped sourdough pro­ baking were purchased from Ubani market in Umuahia, Abia state,
ductions (Gänzle & Zheng, 2019). Back slopped spontaneous sourdough Nigeria.
is generally less elastic, cohesive and more difficult to handle compared
to the gluten containing counterparts (Ronds et al., 2017). Certain Sample preparation
technological and nutritional functional ingredients are therefore added
like cereal hydrocolloids, glutathione, fruit or vegetable fibre, flax, Pearl millet flour
psyllium modified starches and proteins (Smidova & Rysova, 2022) to The millet grains were sorted, washed, steeped for 24 h and drained
achieve sufficient improvement on their bread volume, crumb softness, thereafter. Steeped water was changed at 12 h interval to avoid
shelf life and acceptability (Azizi et al., 2020; Culetu et al., 2021). Be­ fermentation. The steeped grains were then oven dried using a Memmert
sides, modified proteins of maize, sorghum and rice have been exten­ oven at 60 ◦ C for 2 h and milled. The flour was obtained by sieving
sively executed recently through heating, freezing and extrusion to through 0.25 mm sieve mesh and packaged in airtight container for
modify their protein structure and functionality (Gómez & Martínez, baking.
2016; Mirmoghtadaie et al., 2016) for better sourdough improvement.
Sourdough spontaneous back slopping fermentation process is a Bambara nut flour
continuous daily refreshment (Gocmen et al., 2007) of the traditional The seeds were steeped as in pearl millet, hand-dehulled, boiled for
sourdough with active state of the microorganisms from the flour (for 10 min (1:4 beans to water), drained, oven dried at 50 ◦ C for 9 h, milled
acidification) and water to obtain “sponge” or “mother dough” and sieved into flour using 0.25 mm sieve to obtain the flour (Olaoye
Refreshment intervals of longer than three days acidify the dough and et al., 2006). The flour was also packaged in air container for baking.
may change the microbial ecosystem (Wing, 2018). The intervals may be
reduced to alter the ratio of yeasts to lactobacilli (Nanna et al., 2006) to Preparation of soybean paste
increase the rate of gas (CO2) production, called acceleration or reduce The method of Okwunodulu et al. (2017) was used with slight
the amount of starter in the final dough to obtain a satisfactory rise modification. Soybean was sorted, cleaned, steeped for 12 h in tap
during proof. water, boiled for 20 min, drained, cooled to room temperature and
As the flour contacts water, with sufficient time, temperature and manually dehulled. The hulls were removed by water floatation and the
refreshment, the cereal microbes and mother dough amylase enzymes soy cotyledons obtained were milled (2.7 parts of water to one part
break down starch to sugars (glucose and maltose) which the yeast soybean [v/w]) in a variable speed kitchen blender, Power Delux,
ferment to produce carbon dioxide that leavens the dough. The yeasts (Model PDB-8231-F) to obtain fine soybean paste. A part of the paste
also impart flavour through alcohol and ester formation (De Vuyst et al., was dried in oven (Model No TO008GA-34, AKAI-Tokyo Japan) to
2016). The LAB ferments free maltose and monosaccharides to lactic constant weight for functional analyses.
acid and/or acetic acid (Gänzle & Gobbetti, 2013) and further aid sec­
ondary proteolysis of oligopeptides (optimal pH 6.0–8.0) by the intra­ Sourdough preparation and bread baking
cellular peptidases to accumulation amino acids in the sourdough Back slopping method of (Corsetti & Settanni, 2007) was adopted for

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I.N. Okwunodulu et al. Food Chemistry Advances 4 (2024) 100633

the sourdough preparation. Each 200 g of composite flours of Foam capacity (FC). The FC was determined using the method described
millet-Bambara nut and millet-soybean (50 % of each flour type) was by Wu et al. (2009). The FC was calculated as shown:
separately mixed with the following ingredients, 200 g sourdough
V2 − V1
starter (first natural fermented dough which contains LAB and yeast), 2 Foam capacity (%) = × 100
V1
g salt, 100 ml warm water, 30 g sugar, 10 g margarine and 50 g egg. Both
formulations were separately kneaded and refreshed at every 12 h in­ where: V1, initial volume of the concentrate suspension before blending;
tervals, at room temperature (23–26 ◦ C) with 50 g of flour type and 25 V2, Final volume after blending + foam.
ml water (back slopping) for four days to maintain the active state of the
fermenting microorganisms. Each stable “sponge” or “mother dough“ Foam stability (FS). This was obtained by recording the time it took for
produced thereafter was rolled on a wooden table with wooden roller, the foam obtained above to collapse
cut (equal sizes) with stainless steel knife, proofed (1 h) in pans on a
proofing table and baked at 250 ◦ C for 35 min in an electric oven (Model
Emulsion activity (EA). The EA was determined using the method
No TO008GA-34,AKAI-TOKYO JAPAN). The microorganisms in the
described by Neto et al. (2001). 5 ml portion of protein solution (1%
mother dough and refreshing flour fermented the dough during proof­
w/v) was homogenized with 5 ml of soybean oil. The emulsions were
ing. Wheat flour (300 g), 100 ml water and 100 ml yeast were mixed
centrifuged at 1100 per gram for 5 min. The height of the emulsified
with same quantity of ingredients as in sourdough bread, kneaded into
layer and that of the total contents in the tube was measured. The
dough manually in a stainless steel bowl, cut as above, proofed in pans
emulsion activity (EA) was calculated thus;
and baked as above (control sample). The baked breads were allowed to
cool at room temperature, packed with low density polyethylene and Heightofemulsifiedlayer
EA (%) = × 100
refrigerated for analyses. Heightoftotalcontentofthetube

Emulsion stability. Emulsion stability (ES) was determined by heating

Analysis the emulsion at 80 ◦ C for 30 min before centrifuging at 1100 × g for 5
min. The ES was calculated thus:
Proximate composition
Moisture content (MC), dry matter (DM), ash, crude protein (CP), Height of emulsified layer after heating
ES (%) = × 100
crude fibre (CF), fat and carbohydrate content of the flour and the bread Height of emulsified layer before heating
samples were analyzed using the protocol of Onwuka (2018). The en­
ergy values were obtained by calculation using the Atwater general Mineral and vitamin composition of the breads
factor system (AGFS) described by AOAC (2010). Energy values of the
breads were obtained by adding the products of protein, fat, and car­ Calcium. Calcium content of the samples was determined by versanate
bohydrate values with their respective physiological fuel value of 4, 9, EDTA complexiometric titration as described by AOAC (2010). 20 ml of
and 4 kcal/g and that of fibre as shown: sample digest (2 g of the raw samples dissolved in 10 ml of diluted 2 M
HCL solution and filtered through Whatman filter paper) treated with
Energy values kcal) = (% CP x 4) + (% CF x 9) + (% CHO x 4) + (% fibre x2). the masking agents (hydroxylamine hydrochloride, sodium cyanide and
sodium potassium ferrocyanide) and 20 ml of ammonia was added to
where: CP = Crude protein, CF = Crude fat and CHO = Carbohydrate values.
adjust the pH to 10.0 (a point at which calcium forms complexes with
EDTA). The mixture was titrated against 0.02 N EDTA solution to a
permanent deep blue colour end point. A pinch of Eriochrom black T
Functional properties of the flour samples indicator was added, and shaken very well before titration. The result
obtained indicated the amount of combined concentrations of calcium
Bulk density. The method of Onwuka (2018) was adopted. 10 ml and magnesium because Ca2+ and Mg2+ form complexes at pH 10.0
measuring cylinder was weighed and separately filled with each flour with EDTA. Therefore, another titration as above was conducted to
sample. The bottom of the cylinder was tapped gently until no further determine calcium alone by adjusting the pH of the digest to 12.0 with
reduction of the sample level. The new volume was recorded and used to 10 % NaOH and selechrome dark blue (Calcon) as an indicator in place
calculate the bulk density as shown: of Eriochrome black T. A reagent blank was titrated to serve as a control.
( / This experiment was carried out in triplicated and the average reading
Weightof sample(g) was used. The calcium content was calculated thus:
BD g ml) =
Volume of sample (ml)
100 Vf
Calcium (mg / 100 g) = × EW × N × ×T− B
Water absorption capacity (WAC). The method as described by Onwuka W Va
(2018) was used to determine the WAC of the flour samples. The WAC where: W, Weight of sample analyzed; EW, Equivalent weight; N,
was calculated using the relationship: Normality of EDTA; Vf, Total volume of extract; Va, Volume of extract
V1 − V2 titrated; T, Titre value of sample; B, Titre value of blank.
WAC (g / g) = × 100
W
Phosphorus. This was determined using Molybdovanadate colorimetric
where: V1, Initial volume of water before centrifugation; V2, Final vol­ method described by James (1995). Two (2) mg of each sample were
ume of water after centrifugation; W, Weight of sample. dispersed separately in a 50 ml volumetric flask, each mixed with two ml
of Molybdovandate solution and allowed to stand for 15 min at room
Oil absorption capacity (OAC). Also, the method described by Onwuka temperature for colour development. Same treatment was given to 10 ml
(2018) was used here and OAC was calculated as: each of distilled water and standard phosphorus solution in flasks to
V1 − V2 × 100 serve as reagent blank and standard, respectively. They were made up to
Oil absorption capacity (g / g) =
W 20 ml mark with distilled water and absorbance was taken with Jenway
electronic spectrophotometer at a wave length of 420 nm and the re­
where; V1, Initial volume of oil before centrifugation; V2, Final volume agent blank at zero. Phosphorus content was calculated as shown:
of water after centrifugation; W, Weight of the sample.

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I.N. Okwunodulu et al. Food Chemistry Advances 4 (2024) 100633

Table 1
Proximate composition of the flours, soybean paste, sourdough breads and energy levels of breads (%).
Samples MC DM Ash CP CF Fat CHO EV (Kcal)
b b c b b b b
Millet 9.50 ± 0.02 90.50 ± 0.02 0.67 ± 0.04 12.56 ± 0.18 0.88 ± 0.06 2.05 ± 0.10 74.33 ± 0.08 367.77b ± 0.01
Bambara 7.71c ± 0.02 92.84a ± 0.02 1.35b ± 0.01 7.90c ± 0.08 1.41a ± 0.11 3.92a ± 0.24 78.26a ± 0.44 382.74a ± 0.01
Soybean 48.28a ± 0.57 51.72c ± 0.06 2.46a ± 0.07 24.43a ± 0.11 1.10b ± 0.20 3.90a ± 0.07 19.83c ± ± 0.34 214.34 ± ± 0.01
WB 34.71a ± 0.13 65.29b ± 0.13 2.10b ± 0.00 10.54ab ± 0.12 0.84c ± 0.01 4.87b ± 0.04 46.95d ± 0.23 275.47 ± 0.01
MBSB 33.37b ± 0.66 66.63a ± 0.66 1.93c ± 0.01 9.84c ± 0.08 1.17a ± 0.02 6.35a ± 0.07 46.82d ± 0.56 294.13 ± 0.01
MSSB 33.47b ± 0.02 66.54a ± 0.02 2.15a ± 0.02 10.36ab ± 0.03 1.08b ± 0.02 3.78c ± 0.00 49.69b ± 0.06 276.38 ± 0.01

Values are means of triplicate determinations ± standard deviation. Values with different superscripts within the column are significantly different (p < 0.05). MC-
moisture content, DM-dry matter, CP-crude protein, CF-crude fibre and CHO = carbohydrate. WB = Wheat bread, MBSB ¼ millet and Bambara nut sourdough bread
and MSSB = millet and soybean paste sourdough bread.

au C Vt calculated as shown:
Phosphorous (mg / 100 g) = 100 × × ×
w as Va
100 × Vf × T
Vitamin C (mg / 100 g) = 0.88 ×
where; w, weight of ash sample; au, absorbance of test sample; as, 10 20 1
absorbance of standard phosphorus solution; c, concentration of stan­
where; Vf, Volume of extract; T, Sample titre—blank titre.
dard phosphorus solution; Vt, total volume of extract (filtrate); Va,
Volume of extract analyzed.
Physical characteristics of the breads

Iron. Atomic absorption spectrophotometer (AAS) was used to deter­

Oven spring. This was calculated by finding the difference between the
mine the iron concentration of the samples as described by James
heights of the dough before baking and that of the loaf after baking.
(1995). The sample digest was aspirated into a flame which converted
them to free atom vapour. A monochromatic light source was directed
Loaf volume. This was calculated by multiplying. the height, length and
through the flame and the amount of radiation of specific energy was
width of the breads measured with slide rule.
recorded. From this a calibration graph was prepared for the iron from
where the amount in each sample was read. The final computation was
based on this relationship; Loaf weight. This was determined by weighing the loaf directly on the
weighing balance Onwuka (2018).
C×V
Weight of sample used Specific loaf volume. This was calculated as a ratio of loaf volume of each
bread sample and their corresponding weights as shown:
where; C, Concentration of the metal from the sample detected by AAS;
V, Volume of which the solution was diluted. ( / ) Loaf volume (cm3 )
Specific loaf volume cm3 g =
Loaf weight (g)
Vitamins B1. The spectrophotometric method described by AOAC
(2010) was used. 5 g of each sample was weighed and homogenized with Sensory characteristics. The organoleptic attributes of the bread samples
50 ml of 1 N NaOH. The homogenate was filtered and the filtrate ob­ were evaluated as described by Iwe (2002). A total of 20 semi-trained
tained was used for analysis. An aliquot (10 ml) of the filtrate was panelists randomly selected male and female volunteers aged between
treated with equal volumes of 0.1 N K2CrO7 solutions in a flask. Standard 18 and 22 from Michael Okpara University of Agriculture, Umudike
thiamine solution was prepared and diluted to a chosen concentration were used. And informed consents had been obtained from the panelists
(0.5). An aliquot of standard thiamine solution was also treated with 10 before the sensory experiments were carried out without coercion to
ml of dichromate solution. The absorbance of the sample and the stan­ participate.The coded samples were presented to them in similar plates
dard solutions were measured in a spectrophotometer at a wavelength of inside a well-lighted room along with a bottle of water each to rinse their
360 nm with the reagent blank to be used to calibrate the instrument at mouths before and after tasting each sample. They were instructed to
zero. The thiamine content was calculated using the formula; taste each sample and score them based on taste, texture, appearance,
100 × Au × C × Vf × D aroma and general acceptability. The scoring was based on 9-point
Thiamine (mg / 100 g) = Hedonic scale where 9 = like extremely, 5 = neither like nor dislike
W As Va
and 1 = dislike extremely.
where; W, Weight of sample analyzed; Au, Absorption of sample; As,
Absorbance of standard thiamine solution; C, Concentration of standard Statistical analysis
thiamine solution; Vf, Total volume of filtrate; Va, Volume of filtrate
analyzed; D, Dilution factor where applicable. Data obtained were subjected to analysis of variance (ANOVA) of a
completely randomized design (CRD) using the SPSS (version 22) for
Vitamin B2. Same procedure for thiamine was used here except that personal computers. Treatments were separated using Duncans Multiple
standard riboflavin solution was used in place of thiamin. Range Test at 5 % confidence level (p < 0.05). Pearson correlation co­
efficient was used to correlate the values of physical and chemical pa­
Vitamin C. The method used was as described by AOAC (2010). Ten rameters of the sourdough breads.
grams (10 g) of the sample was extracted with 50 ml EDTA/TCA
extracting solution for 1 h and filtered through a Whatam filter paper
into a 50 ml volumetric flask and made up to mark with the extracting
solution. 20 ml of the extract was pipetted into a 250 ml conical flask
and 10 ml of 30 % KI and 50 ml of distilled water were added. This was
followed by 2 ml of 1 % starch indicator and the mixture was titrated
against 0.01 ml CuSO4 solution to a dark end point. Vitamin C was

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I.N. Okwunodulu et al. Food Chemistry Advances 4 (2024) 100633

Results and discussion (especially MSSB) for low income earners in developing countries who
cannot afford meat and fish. Also, a possible economic means of alle­
Proximate composition of the flours, soybean paste and sourdough bread viating protein-energy malnutrition among children and elderly that
samples will relish on the SBs. Millet and Bambara nut are easily grown in the
tropics unlike wheat that is expensively imported.
Moisture content (MC)
The MC is a measure of the water content and storage stability index Crude fiber (CF)
of the sourdough breads (SBs). The proximate results (Table 1) indicated The CF is indigestible carbohydrate that helps in easy evacuation of
slight MC variations (p < 0.05) between the two sourdough bread (SB) stool, weight management, a minor energy source and maintains in­
samples despite the significant (p < 0.05) MC variations between millet, testinal health. Bread is relished by people of all ages and sex, most
Bambara nut flours and soybean paste. This notwithstanding, the MC especially the elderly, children and recovering patients who need fifer
(33.47 %) of sample MSSB (millet-soybean) is slightly higher (33.47 %) diets The WB is low in CF, but the blending positively improved (p <
than 33.37 % from MBSB (millet Bambara nut SB). Higher MC (48.28 %) 0.05) the entire SB CF. Significantly (p < 0.05) higher CF of MBSB (1.17
and protein (24.43 %) of soybean paste may be responsible. Besides, the %) than MSSB (1.08 %) was observed. However, this could stem from
paste may have contained more hydrophilic molecules than Bambara higher fibre content of Bambara nut flour (1.41 %) than soybean paste
nut. Higher MC (34.71 %) of WB (control) than the SB samples may (1.10 %). This analogized with the report that Bambara nut contains
signify higher water absorption capacity of wheat flour than those of high fibre (Alhassan et al., 2015). The CF improvement in this study
millet and Bambara nut. The WB may likely be softer, easier to masti­ compared to the control (0.84 %) becomes a welcome development
cate, takes relatively longer time to stale than the SB samples but may be considering numerous health benefits associated with fibre. Amidst the
more susceptible to microbial growth depending on storage conditions. improvement, the CF of the SBs (1.08–1.17 %) were lower than
The MC values in this study were higher than 4.23–15.5 % reported by 1.76–3.62 % reported by Adisa et al. (2019) probably due to inclusion of
Adisa et al. (2019) from wholegrain millet SB and 12.67–14.13 % for the husk which has high fiber content, and 2.05–2.19 % for bread from
wheat-pigeon pea composite flour bread (Ajayi et al., 2019). May be composite flour (Ajayi et al., 2019).
grain type and their proportions in the blends used may prove the dif­
ference. Also, the higher fibre content in the wholegrain millet may have Fat
contributed to the lower MC. Fat is a source of fat soluble vitamins and one of the major energy
substrate needed for activity. Depending on the body composition, ac­
Dry matter (DM) tivity and amount consumed over time, the excess may result in obesity.
It is of worthy to note that MC which correlates inversely with DM, In foods, depending on storage conditions could initiate off odour and
an index of higher nutrient content, had similar result trend with MC. flavour through rancidity. There was significant (p < 0.05) higher fat
The only exception is that the DM of all SBs is higher than WB which content of MBSB (6.35 %) than MSSB (3.78 %) and the control (4.87 %).
could be attributed to its higher MC. More so, Bambara nut may have Therefore, MBSB had fat improvement (p < 0.05) unlike MSSB
contained more DM that influenced positively the MBSB. compared to control. Higher fat content of millet (2.05 %) and Bambara
nut (3.92 %) composite flour than those of millet and soybean (3.90 %)
Ash may have been the major cause. Despite the improvement, the fat con­
Blends of millet-Bambara nut and millet-soybean are application of tent of the SBs in this study (3.78–6.35 %) was lower than 7.15–12.3 %
cereal-legume technology for nutrient complementation. Millet is low in reported by Adisa et al. (2019), but higher than 2.82–3.69 % reported by
ash, fibre and fat in this study, but higher in soybean paste and Bambara Ajayi et al. (2019) for composite flour breads. Though the entire bread
nut flour. Ash, an estimation of the total mineral content of food, may scare those susceptible to obesity and obesity related diseases as
improved significantly (p < 0.05) in both SBs due to the blending with well as health and weight conscious persons, they will be desired by
likelihood of improving well-defined biochemical functions of the con­ those living active lives.
sumers mostly elderly and children who relish on them. Both need
mineral to prevent osteoporosis in adult and rickets in infants. Sample Carbohydrate
MSSB had higher ash content (2.15 %) than MBSB (1.93 %) and control There was no significant (p > 0.05) carbohydrate content between
(2.10 %) which could be associated to higher ash content of soybean the control (46.95) and MBSB (46.82 %) but both were significantly (p <
(2.64 %) than Bambara nut (1.35 %). There was mineral improvement 0.05) lower than that of MSSB (49.69 %). This indicated that only
only in MSSB compared to the control which could be traced to soybean. sample MSSB had carbohydrate improvement despite higher carbohy­
This notwithstanding, ash content of the entire SBs (1.93–2.15 %) was drate content of Bambara nut (78.26 %) than soybean paste (19.83 %).
within 2.39–3.08 % reported by Adisa et al. (2019) for whole millet and Similar trend was recorded by Ajayi et al. (2019) for composite flour
2.25–2.80 % for complementary blends of soybean, breadfruit and breads. Carbohydrate values of the entire SBs (46.82–49.69 %) were
plantain by Okwunodulu et al. (2020). Mineral content variations of the lower than 63.7–73.3 % reported by Adisa et al. (2019) and 65.82–68.84
blending materials may have contributed to the variations. Despite % from Ajayi et al. (2019). Carbohydrate is one of the major energy
these, the entire SBs ash met the recommended ash value (<3) by substrates which may predisposed the SBs as moderate energy source
FAO/WHO (1991). depending on the quantity of other energy substrate in them. Due to the
low carbohydrate content of the SBs, it could be desired by the obese and
Crude protein (CP) weight conscious persons.
Protein is essential nutrient for body growth, repairs and energy
substrate. Sample MSSB (10.36 %) had higher (p < 0.05) protein content Energy values (EV)
than MBSB (9.84 %) and is traceable to higher CP of soybean (24.43 %) The significant (p < 0.05) EV variations among the breads could be
than Bambara nut (7.90 %) flour. But both are significantly lower than traced to Bambara nut inclusion which contained the highest energy
the control (10.54 %) which may portend superior CP content of the substrates (carbohydrate and fat) compared to other flours. Hence the
wheat than the flours in composite flours used. Adisa et al. (2019) and highest EV value of sample MBSB (294.13 Kcal) and least from sample
Ajayi et al. (2019) reported respective higher protein content of WB (275.47 Kcal). The least EV of WB could be associated to absence of
11.5–15.3 % and 10.19–14.21 % than those obtained in this study. Flour Bambara nut as composite flours improve the essential nutrients more
types and proportion used must have been the major cause of the vari­ than the wheat flour. Therefore, the sourdough breads in this study
ations. With this, the SBs could be an economic protein source could be a better energy source than WB especially infants who are

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I.N. Okwunodulu et al. Food Chemistry Advances 4 (2024) 100633

Table 2
Functional properties of the flours and soybean paste powder.
Samples BD (g/ml) WAC (g/g) OAC (g/g) FC % FS % EA % ES%
a b b b a a
Millet 0.77 ± 0.04 141.59 ± 1.79 146.64 ± 0.30 12.45 ± 0.57 1.83 ± 0.18 40.65 ± 0.21 35.38a ± 0.11
Bambara 0.43b ± 0.06 174.51a ± 0.12 107.46c ± 0.18 14.53a ± 0.11 2.85b ± 0.14 20.28b ± 0.32 28.60b ± 0.21
Soybean 0.59ab ± 0.08 138.86b ± 0.12 204.20a ± 0.14 7.48c ± 0.04 19.83c ± 0.46 10.78c ± 0.11 12.50c ± 0.28

Values are means of triplicate determinations ± standard deviation. Values with different superscripts within the column are significantly different (p < 0.05). BD -
bulk density, WAC - water absorption capacity, OAC-oil absorption capacity, FC - foam capacity, FS- foam stability, EA-emulsion activity and ES-emulsion stability.

Table 3
Minerals and vitamin composition of the bread samples (mg/100 g).
Samples Calcium Phosphorous Iron Vitamin B1 Vitamin B2 Vitamin C

WB 42.85e ± 0.07 59.17d ± 0.61 1.17a ± 0.01 0.23c ± 0.01 0.17d ± 0.00 7.62d ± 0.02
MBSB 51.77b ± 0.09 68.50a ± 0.14 1.06b ± 0.00 0.26b ± 0.00 0.19bc ± 0.00 8.27c ± 0.04
MSSB 49.42c ± 0.25 63.64c ± 0.23 0.78d ± 0.00 0.27a ± 0.00 0.23b ± 0.00 9.53a ± 0.11

Values are means of triplicate determinations ± standard deviation. Values with different superscripts within the column are significantly different (p < 0.05). WB =
Wheat bread, MBSB ¼ millet and Bambara nut sourdough bread and MSSB = millet and soybean paste sourdough breads.

active and those engaged in energy sapping jobs. Foam is a colloidal of many gas bubbles trapped in a liquid or solid
thereby improves textural consistency of foods. Higher FC is a desired
flour attribute in baking due to its high percentage yield of porous
Functional properties of the flours and soybean paste powder
structures (low MC) and their stability amidst processing stress in the
production of varieties of baked products like cakes, muffins, bread and
Bulk density (BD)
as functional agents in other food formulations (El-Adawy & Taha,
The BD gives the idea of the flour porosity or the packing factor
2001). It is interesting to note that soybean powder with highest protein
depending on the flour particle sizes. The BD decides weight and choice
content (Table 3) had the least FC while Bambara nut flour with the least
of packaging materials (Onimawo & Akurbor, 2005). The results
had the highest FC. The ability of protein to trap air bubbles, not
(Table 2) showed significant (p < 0.05) BD variations among the flour
quantity present, may clear the doubt. Higher FC of Bambara nut flour
samples with millet flour having the highest value (0.77 g/ml) and
(14.53 %) than millet (13.45 %) and soybean paste (7.48 %) may be due
Bambara nut the least (0.43 g/ml). This is an indication of the appre­
to low MC as in Table 3. Therefore, Bambara nut flour may likely
ciable variability in the BD components of the millet, Bambara nut and
enhance positively the textural consistency of MBSB than MSSB in this
soybean which in turn influence positively the physical properties of the
study depending on the stability of the foam.
breads. The BD variability could be pinned to differential particle sizes
which in part could be due to differential hardness of their endosperm.
Foam stability (FS)
However, the hardness couldl be substantiated by the discrepancies
The FS points to the ability of protein to stabilize against the gravi­
between their carbohydrate and protein content. Higher values of all the
tational and mechanical stress. Therefore, FS is a function of protein
physical properties of MSSB than MBSB (Table 4) could also justify the
quality which as well determines the tensile strength of the sourdough.
variations. Higher BD values are therefore desired for better physical
High protein quality of soybean had been advanced which justified
properties of bread.
higher FS of soybean powder (19.83 %) than millet (1.63 %) and
Bambara nut (2.85 %) flours. This could explain the reason why bread
Water absorption capacity (WAC)
sample MSSB had better physical properties than MBSB (Table 4) and
This refers to the amount of water absorbed per gram of the flours
may give the MSSB a harder texture.
depending on their particle sizes (Boye et al., 2011) and carbohydrate
content. It is an indication of reconstitution capability of the flour which
Emulsion activity (EA)
increases with decrease in flour particle sizes. It correlates inversely with
The EA is the ability of flour protein to participate in the formation
MC of the flour and the degree of destruction of the macromolecular
and stabilization of newly created emulsion. An emulsion is an unstable
matrix that entraps large amount of water during processing (Chen &
mixture of two or more immiscible liquids with natural tendency to
Lin, 2002). Higher WAC of Bambara nut flour (174.51 g/g) than millet
separate and reduce its interfacial energy. Millet flour had significant (p
(141.59 g/g) and soybean paste (138.86 g/g) may imply that bread
< 0.05) higher EA (40.65 %) than Bambara nut (20.28 %) and soybean
sample MBSM may likely be softer and may take longer time before
paste (10.78 %). The least EA value of soybean could be due to sprouting
staling than MSSB. High WAC flour is therefore a better property for SB
which hydrolyzed most protein into amino acids. Depending on their
production.
protein quality, millet flour protein may likely be the major contributor
to sourdough elasticity and consistency. Following the trend, MBSB
Oil absorption capacity (OAC)
sourdough may likely be better than that of MSSB.
This is the ability of flours to absorb oil which helps to improve the
mouth feel, flavour and fat soluble vitamins of the SB. Soybean paste
Emulsion stability (ES)
powder with higher OAC capacity (204.30 g/g) followed by millet
This annotates the capability of the flour samples to resist changes in
(146.64 g/g) and Bambara nut (107.46 g/ml) may likely boost the
their physicochemical properties over time. Creaming, flocculation,
acceptability of bread sample MSSB more than MBSB by retaining most
coalescence among others can cause emulsion breakdown. Millet flour
of the fat used in the kneading. This is evidenced in the higher accept­
had significant (p < 0.05) higher EC (35.38 %) than Banbara nut (28.60
able level of MSSB (5.85) than MBSB (5.20) in Table 5. Therefore, flours
%) and soybean paste powder (12.50 %). This trend which is same with
with high OAC are better options for SB baking.
EA may mean that sample MBSB may resist physicochemical changes
during kneading and other processes than that of MSSB. With this,
Foam capacity (FC)
higher (p < 0.05) ES of Bambara nut flour than soybean may explain the
The FC refers to the interfacial area that can be created by protein.

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I.N. Okwunodulu et al. Food Chemistry Advances 4 (2024) 100633

Table 4 major positive impact of soybean in deciding the physical properties of

Physical characteristics of the bread samples. the entire breads. Higher bread volume, weight and specific loaf volume
Samples Oven spring Loaf vol. (cm3) Loaf weight (g) SLV (cm3/g) of MSSB than MBSB may impart be a confirmation (Table 4).
(cm)

WB 4.50a ± 0.07 196.72a ± 237.00a ± 8.30ab ±

Mineral and vitamin composition of the bread samples
0.03 1.41 0.07
MBSB 0.35c ± 0.07 172.13c ± 214.00b ± 8.06b ± 0.08
0.04 2.83 Calcium
MSSB 0.95b ± 0.03 186.88b ± 219.00b ± 8.50a ± 0.07 Micronutrient results (Table 3) indicated significant (p < 0.05) var­
0.62 1.41 iations in calcium content among the SBs with sample MBSB (51.77 mg/
Values are means of triplicate determinations ± standard deviation. Values with 100 g) having the highest value followed by MSSB (49.42 mg/100 g) and
different superscripts within the column are significantly different (p < 0.05). control (42.83 mg/100 g). This is significant (p < 0.05) calcium
WB = Wheat bread, MBSB ¼ millet and Bambara nut sourdough bread and improvement of the SBs compared to the control and could predisposed
MSSB = millet and soybean paste sourdough bread SLV-specific loaf volume. the SBs as better calcium source. Higher calcium value of MBSB than
MSSB may signify higher calcium content of Bambara nut flour than
soybean despite sprouting that improve mineral content of legumes.

Fig. 1. Photos of flours used and the prepared breads: a) millet flour; b) Bambara nut flour; c) soybean flour; d) white wheat flour; e) millet-Bambara nut sourdough
bread; f) millet-soybean sourdough bread.

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I.N. Okwunodulu et al. Food Chemistry Advances 4 (2024) 100633

Calcium improvement could be linked to phytate hydrolysis by phytate Physical characteristics of the bread samples
enzyme of the sourdough during fermentation (Oboh et al., 2003).
Calcium is the most abundant mineral in the body responsible for strong Oven spring (OS)
bones, teeth among others. Its improvement is a positive achievement as The OS is the sudden bread volume increase as a result of sudden
that will predispose SBs, most especially MBSB as a potential candidate temperature rise when the dough is exposed to oven temperature due to
in preventing osteoporosis among the elderly and rickets in infants that increased yeast enzyme activities. It is a function of protein type and
mostly relish on bread. Similar calcium improvement compared to the quality as well as enzyme activity which points to wheat flour (control)
control had been advanced (75.50–116.30 mg/100 g) by Ajayi et al. as superior in baking compared to other composite flours. Physical
(2019) which is higher than the values in this study. Calcium content of property results (Table 4) identified differed significant (p < 0.05) OS of
the SBs in this study was higher than 46.79–48.86 mg/100 g (Adisa the entire bread samples with WB having the highest value (4.50 cm)
et al., 2019). Flour type and fermentation time could be the source of while MBSB had the least (0.35 cm). Superior wheat flour gluten content
variation. and use of specific yeast starin compared to consortium of wild yeast and
LAB from the flours used in the sourdough fermentation validated the
Phosphorous wide OS variation. Furthermore, the wild yeast may have been not
Significant (p < 0.05) phosphorous content variation between MBSB enough to ferment and produce appreciable CO2 to increase the OS of
(68.50 mg/100 g) and MSSB (63.64 mg/100 g) were higher than the the SBs. Again, lactic acid bacteria (LAB) may have dominated and
control (59.17 mg/100 g) indicating influence of cereal and legume produce lactic acid that lowered the dough pH thereby restricting yeast
composite flour blends compared to whole wheat flour. Higher SBs activities. Higher protein content of millet and soybean flours than that
phosphorous values depend on the phosphorus and anti-nutrient levels of millet and Bambara nut (Table 1) may explain higher OS values of
of the composite flours used and may predispose Bambara nut flour as a MSSB (0.95 cm) than MBSB (0,35 cm). High loaf OS is a measure of
better source than soybean. Phosphorous synergizes with calcium to number of yeast presence with increased activities unlike low values
enhance calcium absorption which implies adequate proportion of each indicate dense and compact crumb liable to cracks typical of SB
in the diet. The calcium: phosphorous ratio of the entire breads (Ogunsakin et al., 2015). This is evident in more cracks in sample MBSB
approximated to 1:1 but MSSB have a slight edge over MBSB and con­ (Fig. 1e) than MSSB (Fig. 1f) while WB (Fig. 1d) had none. Protein su­
trol. This fell within 1.5–2.1 range recommended by the nutritionists for periority in part may be the source of the crack variations.
bone health and development (Okwunodulu et al., 2022). This therefore
predisposed SBs especially MSSB than WB for better strong bones and Loaf volume (LV)
development. Cereal-legume blend may have made the difference. The Similar OS trend results were obtained here. Superior loaf volume
SB phosphorous content in this study (63.64–68.50 mg/100 g) is higher (LV) of sample MSSB (186.88 cm3) than MBSB (172.13 cm3) could be
than (7.3–9.1 mg/100 g) reported by Adisa et al. (2019), but lower than due to the higher retention of gas released by the yeast activity during
(75.50–116.30 mg/100 g) for pigeon pea: wheat flour bread. The vari­ sourdough fermentation depending on protein quality. Higher OS of
ations could be linked to the phosphorous levels of the individual flours MSSB could substantiate that. The OS improvement in this study is a
in the blends. welcome development and will attract more sales with income since
consumers purchase breads based on size. The LV values in this study
Iron were lower than 280.72–365.66 cm3 reported by Yusufu and Ejeh
The control sample had higher (p < 0.05) iron content (1.17 mg/100 (2018) and 249.1–500.61 g/cm3 by Adisa et al. (2019) respectively for
g) than MBSB (1.06 mg/100 g) and MSSB (0.78 mg/100 g) which is not SBs from whole wheat-Bambara nut flour and whole grain millet. The
iron improvement. Iron content of the wheat flour may have been su­ variations could be traced to flour type, protein quality and the level of
perior to those of the composite flours or they may have contained more yeast activities.
phytic acid that binds and reduces iron bio-availability. Besides, sour­
dough fermentation and time may not have hydrolyzed enough of their Loaf weight (LW)
phytic acid content. These notwithstanding, both SBs will help alleviate The LW of the control sample (237 g) is significantly (p < 0.05)
incidence of anemia among the consumers over time as they proved to higher than MSSB (219 g) and MBSB (214 g) which had no significant
be good iron source. Iron is the least mineral in this study. Adisa et al. (p>0.05) variation between them. The higher LW of the control could be
(2019) and Ajayi et al. (2019) respectively reported higher iron values of associated with the BD of wheat carbohydrate compared to other flours.
2.72–2.78 mg/100 g and 2.65–3.11 mg/100 g. Similarly, slight higher value of MSSB than MBSB could be traced to
higher BD of soybean flour than Bambara nut flour (Table 2). Weight
Vitamin B1 improvement is not to the advantage of the baker as consumers may not
The B1 content of the SBs improved as evident in the significant (p < be attracted much. The entire SB’s LW were lower than 245.75–246.97 g
0.05) higher value of MSSB (0.76 mg/100 g) and MBSB (0.26 mg/100 g) reported by Yusufu and Ejeh (2018) but higher than 123.03–141.87 g
than the control (0.23 mg/100 g). The improvement could depend on reported by Adisa et al. (2019) may be due to lower BD. Increased
the B1 content of the composite flours and the extent of increase by moisture absorption and low carbon dioxide gas retention that results in
sourdough fermentation (Oboh et al., 2003). The improvement could heavy loaves may be the sources of variations (Olaoye et al., 2006).
also be tied to cereal–legume blending.
Specific loaf volume (SLV)
Vitamin B2 This is the volume of unit mass of the SBs. Sample MSSB had sig­
Just like in B1, there was an improvement in B2 of SB samples nificant (p < 0.05) higher (8.50 cm3/g) SLV than MBSB (8.06 cm3/g)
compared to the control (0.17 mg/100 g) with sample MSSB having a and control (8.30 cm3/g) despite higher oven spring of the control. This
higher value (0.23 mg/100 g) than MBSB (0.19 mg/100 g). could be associated to higher crude protein content of soybean (Table 1)
and its functionality. The protein in MSSB may have retained more
Vitamin C carbon dioxide gas produced by the yeast than the rest thereby caused
Same trend of improvement in B1 and B2 was observed in vitamin C the dough to extend with bigger volume.
which implied that sourdough technology in this study had significant (p
< 0.05) vitamin C improvement probably due to dough fermentation Correlation of values of physical and chemical parameters of the sourdough
and vitamin C content of the composite flours. breads
Pearson correlation of the chemical and physical parameters of the

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I.N. Okwunodulu et al. Food Chemistry Advances 4 (2024) 100633

Table 5 Funding source

Sensory scores of the bread samples.
Samples Taste Texture Appearance Aroma General There was no external financial aid from any source.
acceptability

WB 7.30a ± 7.65a ± 7.10a ± 6.60a ± 7.50a ± 1.10 CRediT authorship contribution statement
1.08 1.09 1.45 1.14
MBSB 4.50b ± 5.15b ± 5.90a ± 4.74b ± 5.20b ± 1.80 I.N. Okwunodulu: Writing – review & editing, Project administra­
2.09 1.18 2.05 1.62
tion, Methodology, Conceptualization. G.N. Egbuta: Writing – original
MSSB 5.20b ± 5.80b ± 6.25a ± 5.65ab ± 5.85b ± 1.42
2.14 1.96 1.97 2.11 draft, Resources, Data curation. F.U. Okwunodulu: Supervision,
Methodology, Formal analysis. C.M. Ojinnaka: Software, Resources. S.
Values are means of triplicate determinations ± standard deviation. Values with
C. Onyeiwu: Visualization, Validation, Resources.
different superscripts within the column are significantly different (p < 0.05).
WB = Wheat bread, MBSB ¼ millet and Bambara nut sourdough bread and
MSSB = millet and soybean paste sourdough bread. Declaration of competing interest

sourdough breads indicated that calcium and phosphorous had negative The authors declare that there are no conflicts of interest.
correlation (p < 0.01) with loaf volume, loaf weight and specific loaf
volume. This implied that both reduced the sourdough physical prop­ Data availability
erties most especially the specific loaf volume. Conversely, vitamin C
had positive correlation (p < 0/01) with the physical properties indi­ No data was used for the research described in the article.
cating their increase in physical properties. Therefore, addition of
composite flour or any ingredient containing more vitamin C with
moderate calcium and phosphorous may likely enhance the physical Acknowledgement
propertied of the sourdough breads and will in turn enhance their
acceptability. Authors are grateful to the Department of Food Science and Tech­
nology Department for provision of the analytical reagents used and
Sensory evaluation of sourdough bread research space.

Sensory scores (Table 5) established that the control was liked very Supplementary materials
much (7.50), sample MSSB liked slightly (5.85) and MBSB neither liked
nor disliked (5.20). There were no significant (p > 0.05) acceptability Supplementary material associated with this article can be found, in
variation between the SBs. This notwithstanding, sample MSSB had the online version, at doi:10.1016/j.focha.2024.100633.
slight higher value in all the attributes than MBSB but there were no
significant (p > 0.05) variations between them except in aroma. Slight References
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