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Received: 8 July 2022    Revised: 22 August 2022    Accepted: 24 August 2022

DOI: 10.1002/fsn3.3060

ORIGINAL RESEARCH

Formulation optimization of functional wheat bread with low

glycemic index from technological and nutritional perspective

Neda Mollakhalili-­meybodi1,2  | Mohammad Hassan Ehrampoush3 |

Bahador Hajimohammadi2,4  | Mohammad Hassan Mosaddegh5

1
Department of Food Science and
Technology, School of Public Health, Abstract
Shahid Sadoughi University of Medical
Inclusion of prebiotic compounds as indigestible dietary fiber in wheat bread has
Sciences, Yazd, Iran
2
Research Center for Food Hygiene and
grown rapidly considering the increased public awareness about their impact on health.
Safety, Shahid Sadoughi University of However, through their incorporation, the technological characteristics may adversely
Medical Sciences, Yazd, Iran
3
be influenced by gluten dilution impacts. This study was done to evaluate the impacts
Environmental Science and Technology
Research Center, Department of of long chain, native and short chain inulin (L-­, N-­, and S-­type inulin, respectively) at 8%,
Environmental Health Engineering, 10%, 12%, 14%, and 16% w/w as Inulin Reconstituted Wheat Flour (IRWF) with similar
School of Public Health, Shahid Sadoughi
University of Medical Sciences, Yazd, Iran gluten: carbohydrate ratio of wheat flour (at 10%, 12.5%, 15%, 17.5%, 20% w/w) on tech-
4
Department of Food Hygiene and Safety, nological and nutritional value of wheat bread. Results indicated that despite no gluten
School of Public Health, Shahid Sadoughi
dilution induced by IRWF supplementation, technological characteristics were adversely
University of Medical Sciences, Yazd, Iran
5
Department of Pharmacology, School of influenced especially at higher substitution level of L-­type-­containing formulations which
Pharmacy, Shahid Sadoughi University of is attributed to their higher water absorption index (WAI). Reversely, the nutritional
Medical Sciences, Yazd, Iran
value was positively influenced in which the lowest hydrolysis index (26.64%); predicted
Correspondence Glycemic Index (51.93%) and fructan loss content (25.42%) were found at L-­type inulin-­
Neda Mollakhalili-­meybodi, Shahid
Sadoughi University of Medical Sciences, containing IRWF at the highest substitution level (20% w/w). As the nutritional value
Gomnam Blvd, P.O.Box 8915173160, of wheat bread as staple foodstuff is important, optimizing the bread-­making process
Yazd, Iran.
Email: neda_mabodi@yahoo.com to decrease all reverse impacts induced by L-­inulin-­type inclusion seems to be required.

Funding information KEYWORDS

Shahid Sadoughi University of Medical inulin, nutritional, reconstitution, supplementation, technological, wheat bread
Sciences, Grant/Award Number: 6740

1  |  I NTRO D U C TI O N (sourdough) (Bansal & Kapoor, 2015). Wheat flour is preferred to be

Bread is recognized as one of the main foods consumed globally.
Regarding World Health Organization (WHO) recommendation, the
per capita consumption of bread is estimated at about 250 g per day
(Serrem, 2011). However, it is consumed at a higher ratio especially
in cereal-­based diet societies (developing countries). In Iran, its con-
sumption rate is estimated at about 320 g per day.
Bread is prepared in different shapes, sizes, and textures through
the world using the combination of flour, water, salt, and yeast
used in bread-­making process regarding its distinctive viscoelastic
characteristic to provide gluten network to restore gases produced
throughout the bread-­making process. Despite the nutritional and
technological desirability of wheat bread, its high glycemic index (GI)
will be a threat for health, especially in people who suffer from type
II diabetes (Borczak et al., 2018).
GI is calculated by the ratio of area under curve (AUC) in re-
sponse to a test and reference food consumed by one on different
days and standard situations. AUC is an indicator of blood glucose

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
provided the original work is properly cited.
© 2022 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.

Food Sci Nutr. 2022;00:1–11.  |

wileyonlinelibrary.com/journal/fsn3     1
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2      MOLLAKHALILI-­MEYBODI et al.
response (Reka et al., 2017). In this regard, foods with GI lower
than 55, 55–­69, and above 70 are known as low, middle, and high
GI foods, respectively (Rahelić et al., 2011). The GI of white wheat
bread is 100 (Schneider et al., 2015). Low GI food consumption is
preferred by those who are trying to lower their blood glucose level,
for example, diabetic patients.
Carbohydrate fractions of food supplies like cereal differ in
size, shape, chain length, amylose: amylopectin ratio, and branch-
ing degree (Gourineni et al., 2017; Zhang & Hamaker, 2009). The
postprandial glucose response of this fraction is determined by
its digestibility ratio. In this regard, starch is classified as rapidly
and slowly digestible starch abbreviated as RDS and SDS, respec-
tively, and resistant starch (RS) (Chung et al., 2009). High incorpo-
ration of RS and SDS are claimed to have low GI (Bharath Kumar &
Prabhasankar, 2014).
Regarding the high GI of white wheat bread as a staple foodstuff,
any attempt to reduce the portion of carbohydrate and its digest-
ibility is appreciated especially for people who suffer from obesity,
diabetes, and insulin resistance. Using protein, fat and fiber are
considered as the main strategies to reduce the GI of foods rich in
carbohydrates like wheat bread (Wee & Henry, 2020). Considering
the importance of carbohydrates in the technological characteristics
determination of wheat bread, using dietary fiber is recommended
to be useful in the formulation of low GI wheat bread.
Prebiotics as indigestible carbohydrates act as bifunctional
agents. In other words, their incorporation in food formulations,
however decreases their postprandial glucose response, and en-
hances the growth/activity of probiotic bacteria in the gastrointesti-
nal tract (Carlson et al., 2017; Mohebbi et al., 2018). Among different
compounds known as prebiotic, Inulin as a polymeric compound of
fructose with a polymerization degree of 2–­60, joined by β1-­2 link-
ages is one of the most commonly used compounds in food formula-
tions (Shoaib et al., 2016). Inulin is categorized considering its degree
of polymerization (DP) as short chain (S) (DP ≤10), long chain (L) (DP
≥23), and native inulin (N) (DP: 2–­60).
Inulin is found naturally in some plants, microorganisms or pro-
duced enzymatically from sucrose syrup using fructosyl transfer-
ases (Mollakhalili-­Meybodi et al., 2021). Regular intake of inulin as
a prebiotic compound can stimulate the growth of bifidobacteria,
produce short-­chain fatty acids (SCFAs) and lower the intestinal pH,
thus reducing the activity of pathogenic bacteria, cholesterol ab-
sorption, and colon cancer risk, alongside the reduction of GI in its
incorporated food supply (Sirbu & Arghire, 2017). However, keeping
the intact structure of inulin through food formulation and process-
ing is prerequisite to be efficient as prebiotic; the technological and
organoleptic characteristics of wheat bread should not be also neg-
atively affected to guarantee its regular intake by the consumers. As
the dietary fiber incorporation can adversely impact the technolog-
ical characteristics of wheat bread through gluten dilution impact,
reconstitution technique was used in this study. In other words, the
quantities of starch and gluten were determined in wheat flour and
used at the same ratio in Inulin Reconstituted Wheat Flour (IRWF)
(Arya et al., 2016).
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Considering the importance of polymerization degree of inulin
in determination of its technological characteristics efficiency, di-
gestibility and processing susceptibility, the aim of this study was
to assess the impacts of inulin DP and IRWF incorporation level on
technological and in vitro nutritional value of wheat bread.
2  |  M ATE R I A L A N D M E TH O DS
2.1  |  Materials
Inulin (derived from chicory and at S, N, and L types) was provided
by SENSUS (Netherland). All flours had at least inulin content of
more than 99.5% w/w. Wheat flour and vital gluten protein were
purchased from Roshan and Hedia companies (Yazd, Iran), respec-
tively. Wheat flour contained moisture, ash, and protein in quantities
of about 13.95%, 0.23%, and 10.93% w/w, respectively. Yeast was
obtained by Khuzestan Yeast Company (Dezful, Iran) and the other
constituents (salt, sugar, and canola oil) were provided from the local
supermarket. All chemicals including sodium hydroxide, potassium
hydroxide, hydrochloric acid, sodium borohydride, parahydroxy-
benzoic acid, sodium acetate, acetic acid, maleic acid, tris-­maleate
buffer (pH 6.9), D-­fructose, α-­amylase, sucrase, and sodium carbon-
ate were obtained by Merck Company (Germany). Pancreatic ɑ am-
ylase, sucrase, β-­amylase, pepsin, pullulanase, inulinase, invertase,
α-­glucosidase, and α-­cellulose were also provided by Sigma-­Aldrich
company (America).
2.2  |  Method
This study with the aim of studying the potential of Inulin incorpo-
ration in reconstitution of wheat bread from technological charac-
teristics and nutritional value perspective was set in full factorial
experimental design. So, the impact of inulin DP (S, L, and N) and
substitution level of inulin reconstituted wheat flour (IRWF) was
investigated as demonstrated in Table  1. The proportion of gluten
and inulin was similar to the gluten: starch ratio of basis wheat flour
which is 15.33% w/w and 61.32% w/w, respectively.
2.3  |  Farinograph
Brabender farinograph (Brabender, Germany) was used to deter-
mine the quantity of water to reach the consistency of 500 BU ac-
cording to the AACC Method 54-­21A. In this regard, the gluten and
inulin were mixed well with wheat flour into the 300 g mixing bowl of
farinograph which was operated at 30 ± 0.2°C. Farinograph curves
were used for determination of the water absorption (percentage
of water needed to provide a dough consistency of 500 BU), dough
development time (time to reach a consistency of 500 BU), stability
time (time in which the dough consistency is over 500 BU) and falling
quality number were obtained.
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MOLLAKHALILI-­MEYBODI et al.       3

TA B L E 1  Formulations of the present

Independent variables
study on the basis of inulin polymerization
degree and its incorporation level Polymerization Inulin incorporation Gluten incorporation
Number degree of Inulin level (% w/w) level (% w/w) Trial

1 S 8 2 F1
2 S 10 2.5 F2
3 S 12 3 F3
4 S 14 3.5 F4
5 S 16 4 F5
6 L 8 2 F6
7 L 10 2.5 F7
8 L 12 3 F8
9 L 14 3.5 F9
10 L 16 4 F10
11 N 8 2 F11
12 N 10 2.5 F12
13 N 12 3 F13
14 N 14 3.5 F14
15 N 16 4 F15
16 -­ -­ -­ F16

Note: S: (short-­chain inulin with DP ≤10); N: (native inulin with DP range of 2–­60); L: (long chain
inulin with DP ≥23). F16 is the control sample.

2.4  |  Preparation of bread
Bread preparation on the basis of formulations represented in
Table 1 was done as follows with all ingredients added on flour dry
basis. So, active dry yeast, sugar, salt, and canola oil at 2.2%, 0.5%,
1% and 3% w/w were mixed. The water inclusion level was on the
basis of farinograph data calculation. The mixture was then stored
inside the fermentation cabinet for 4 h at 29 ± 0.5°C. The baking pro-
cess was done in a convection oven at 220 ± 10°C. Determination of
proper baking time was according to feasibility of dough separation
from the pan.
2.5  |  Physical characterization of bread
a 5  kg loading cell and cylinder probe (diameter of 43 mm), bread
crumb with dimension of 20 × 20 × 25 mm was pressed up to its half
height at speed ratio of 1 mm/s. The analysis was done in five rep-
licates at room temperature. Texture profile analysis curves were
assessed for hardness, springiness, and chewiness determination
(Mohammadi et al., 2021).
2.6  |  Crust and crumb color estimation
The Hunter Lab instrument was used for color determination of
bread (both crust and crumb). Results are expressed as L* (bright-
ness/darkness), a* (redness/greenness), and b* (yellowness/blue-
ness) indices (Shiri et al., 2021).

2.5.1  |  Specific volume

2.7  |  Sensory evaluation
Rape Seed displacement method was used for volume determination
of loaves according to AACC 109 method 10–­05. After volume and Bread samples were sensory evaluated by nine-­point hedonic scale.
weight determination, specific volume was determined by their ratio In this regard, 1, 2, 3, 4, 5, 6, 7, 8, and 9 were indicators of extremely
as follows: unpleasant, very much unpleasant, moderately unpleasant, slightly
unpleasant, generally acceptable, slightly pleasant, moderately
volume cm3 pleasant, very much pleasant and extremely pleasant bread samples,
( )
Specific loaf volume = (1)
weight (g) respectively. Color, flavor, texture, and overall acceptability were as-
sessed as qualitative parameters by 30 semi-­trained panelists (male:
2.5.2  |  Textural parameters female ratio of panelists was 50:50 with the range of 18–­58 years
old). So, encoded bread samples were served randomly. The pan-
Instrumental texture profile analysis of breads was done and fol- elist's healthiness (with no allergy to gluten protein) and eagerness
lowed using texture profile analyzer (TA 20., KOOPA). Regarding, to be participated were also considered (Menon et al., 2015).
 |
4      MOLLAKHALILI-­MEYBODI et al.
2.8  |  Nutritional value
2.8.1  |  Total sugar (TS)
Total sugar content was quantified as follow (Albalasmeh et al., 2013).
Regarding, 10 μl of sugar extract (by adding 1 ml of 0.005 M acetic
acid to 50 mg bread sample, vortexing for 10 min and centrifuging at
12000 × g for 10 min [Figueroa & Khan, 1991]) was transferred to
5% aqueous solution of phenol (300 μl). After incubation for 5 min,
1.8 ml of concentrated sulfuric acid (18.4 M) was added. They were
vortexed for 30 s and incubated in a water bath at room temperature
for 20 min. Then, absorption was read at 490 nm. Total sugar was
calculated as mg glucose/g extract.
2.8.2  |  Resistant starch (RS)
The RS was estimated on the basis of the method described by
(Reshmi et al., 2017). The protein removal was done by incubation
of 100 mg sample with pepsin solution (40,000 U/ml, 1 g/10 ml KCl-­
HCl buffer) at 40°C for 60 min. Pancreatic α-­amylase solution (40 mg
a-­amylase: 200 U/ml) was used for 16 h at 37°C to hydrolyze starch.
After washing with ethanol (99% v/v) and centrifuging, the superna-
tant pellets were digested with 2 M potassium hydroxide. Incubation
of digested pellet and supernatant was done separately with amylo-
glucosidase (3300 U/ml) and the released glucose was measured
by GODPOD kit and absorbance at 510 nm. The RS and Digestible
starch (DS) were quantified as follows:
RS: glucose (mg) × 0.9 (2)
DS: TS − RS (3)
2.8.3  |  Predicted glycemic index (pGI)
In vitro method was used to determine the pGI on the basis of the
procedure proposed by (Reshmi et al., 2017). Regarding, 100 mg
of ground bread sample was incubated with HCl–­KCl buffer (10 ml
and pH 1.5) and 200 μl pepsin solution (100 mg/ml HCl-­KCl buffer)
for 1  h at 40°C. By adding 200 μl pancreatic α-­amylase solution
(1.5  mg/10 ml phosphate buffer), the pH was increased to 7.8 and
incubated for 45 min at 37°C. To stop the enzyme reaction, Na2CO3
solution was added at 70 μl and using trismaleate buffer at pH = 6.9,
samples were diluted to 25  ml. Then, 5 ml of pancreatic α-­amylase
solution (3 U/5 ml tris-­maleate buffer) was added and incubation was
done at 37°C.
About 1 ml of samples were treated with 3 ml of 0.4 M sodium ac-
etate buffer adjusted at pH equal to 4.75 and 60 μl amyloglucosidase
(3300 U/ml) and incubated at 60°C for 45 min. Afterward, distilled
water was used for the volume adjustment to 10 ml. The solution
(0.1 ml) was transferred into glass test tubes for glucose quantifica-
tion by GODPOD kit and absorbance at 510 nm. The hydrolysis index
(HI) was estimated as the ratio of total released glucose of samples
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and standard glucose (Barine & Yorte, 2016). The pGI of the samples
was calculated on the basis of the following equation (Nathakattur
Saravanabavan et al., 2013);
pGI = 39.71 + 0.549HI (4)
2.8.4  |  Fructan content quantification
The quantification of inulin in bread formulations was done enzy-
matically. In this regard, about 1 g bread sample was dissolved in
80 ml water (80°C). The mixture was filtered and treated with 1 ml
sucrase/amylase solution (sucrase (50 U), β-­amylase (500 U), pullula-
nase (100 U), and maltase (1000 U) in 22 ml sodium maleate buffer
(pH 6.4) to remove sucrose, starch and reducing sugars). Afterward,
alkaline borohydride (10  mg/ml sodium borohydride in 50 mM
NaOH) was added and incubated at 40°C for 30 min and the excess
was removed using acetic acid (100 mM) to reach the pH equal to
4.5. Quantification of fructan was done by fructanase (350 U/ml
exo-­inulinase and 35 U/ml endo-­inulinase) for 20 min at 40°C. The
fructan content (%) was estimated as:
Total fructan ( % ) = A × F × 5 × V × 1.1 ∕ 0.2 × 100 ∕ w × 1 ∕ 1000 × 162 ∕ 180
(5)
where:
A: absorbance of 0.2 ml of the reactive solutions read against the
reagent blank.
F: equal to 54.5, factor used to convert the absorbance value to
μg fructose.
V: volume of used extractant.
M: weight of test portion.
2.9  |  Statistical analysis
Quantitative characteristics were described using mean and stand-
ard deviation. All examinations were done in triplicate. Data analysis
was performed using two-­way anova and Kruskal–­Wallis for para-
metric and non-­parametric tests, respectively, by spss V.21 software
at a significance level of 0.05.
3  |  R E S U LT A N D D I S CU S S I O N
3.1  |  Effect of IRWF on farinograph parameters
The viscoelastic network formation of dough is influenced greatly
by the other ingredients (Meybodi et al., 2019). Wheat flour supple-
mentation with IRWF changes the dough mixing characteristics sig-
nificantly as determined by farinograph. Flour quality is determined
by Farinograph test according to dough physical resistance through
mixing and/or kneading. Farinograph characteristics of wheat flour
and those supplemented with IRWF, at 10%, 12.5%, 15%, 17.5%, and
20% w/w are shown in Table 2.
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MOLLAKHALILI-­MEYBODI et al.       5

TA B L E 2  Farinograph characteristics of
Parameters
wheat flour supplemented with inulin-­
containing reconstituted wheat flour Samples WA (g/100 g) DDT (min) ST (min) FQN (−)
g k i
F1 52.12 ± 0.16 2.23 ± 0.12 9.19 ± 0.03 77.12 ± 0.07h
f k h
F2 54.09 ± 0.07 2.27 ± 0.05 11.09 ± 0.07 81.12 ± 0.10 g
F3 55.74 ± 0.01e 3.01 ± 0.09j 12.11 ± 0.14g 99.65 ± 0.51g
d i f
F4 57.01 ± 0.23 3.64 ± 0.10 13.06 ± 0.01 108.71 ± 0.21f
F5 58.13 ± 0.01cd 4.08 ± 0.17h 14.28 ± 0.08e 112.01 ± 0.62e
e j f
F6 55.65 ± 0.09 3.14 ± 0.12 13.17 ± 0.21 118.35 ± 0.09c
F7 56.17 ± 0.17d 4.53 ± 0.05g 14.08 ± 0.03e 105.08 ± 0.07f
c f d
F8 58.38 ± 0.05 5.17 ± 0.04 15.01 ± 0.23 115.10 ± 0.05d
F9 59.14 ± 0.14c 6.04 ± 0.06e 16.37 ± 0.04c 118.35 ± 0.14c
bc cd b
F10 60.09 ± 0.02 7.13 ± 0.25 17.01 ± 0.08 120.03 ± 0.51c
F11 58.09 ± 0.05d 6.87 ± 0.09d 15.25 ± 0.09d 112.92 ± 0.32e
c c c
F12 58.97 ± 0.08 7.93 ± 0.08 16.48 ± 0.10 115.89 ± 0.43d
F13 60.57 ± 0.05bc 9.03 ± 0.21b 17.28 ± 0.17b 118.16 ± 0.21c
b a a
F14 61.09 ± 0.51 9.43 ± 0.17 18.71 ± 0.21 123.14 ± 0.41b
F15 63.24 ± 0.01a 10.05 ± 0.16a 19.23 ± 0.42a 125.07 ± 0.32a
g k i
F16 51.07 ± 0.23 2.13 ± 0.04 8.73 ± 0.19 63.02 ± 0.16i

Note: Data are reported as average ± standard deviation. The values with the different lowercase
letters in each column mean the significant difference (p < .05).
Abbreviations: DDT: dough development time; FQN: farinograph quality number; ST: stability time;
WA: water absorption.

As demonstrated, wheat flour supplementation with IRWF
influenced the WA content in an inulin DP-­dependent manner
(p  < .05). In other words, while the ability to absorb water is re-
mained quite unaffected at formulation containing 8% w/w S-­t ype
inulin-­containing IRWF (F1) (p > .05), it has been increased in the
presence of 8% w/w L-­t ype inulin-­containing one (F10) compared
to control (F16). The WA enhancement at formulations supple-
mented with IRWF-­containing long chain inulin is mainly attributed
to their ability to provide a gel-­like structure potentially able to
restore water as reported by (Mohammadi et al., 2021). Regardless
of inulin DP, it is clear that supplementation level is also influen-
tial in WA quantity determination of wheat flour which is in the
range of 2.05%–­15.43%, 8.93%–­17.66%, and 15.46%–­23.53%
w/w for S, N, and L inulin-­containing IRWF (at ascending ratio),
respectively. However, this enhancement is more obvious at long-­
chain inulin-­containing formulations. The WA enhancement via
inclusion of different vegetable protein has also been previously
stated which is attributed to their difference in WA capacity and
consequently increased WA value in farinograph plot (Coţovanu &
Mironeasa, 2022).
Dough development time (DDT) is also found to be influenced
by IRWF supplementation. DDT is the required time for flour to be
hydrated, developed, and provided consistent dough equal to 500
Brabender Unit (BU) consistencies. Results indicated significant
(p  < .05) enhancement of DDT at all IRWF-­containing formula de-
pending on polymerization degree of inulin and IRWF supplemen-
tation level (p < .05) (Table 1). DDT enhancement induced by IRWF
inclusion could be attributed to physicochemical characteristics
difference in constituents of IRWF and wheat flour (Paraskevopoulou
et al., 2010). Increased DDT (p < .05) found by increasing the DP of
incorporated inulin in IRWF formulation, is also attributed to higher
susceptibility of L-­type inulin to interact with gluten proteins and
prevent their hydration and development and consequently lead to
DDT enhancement (Graça et al., 2020).
Stability time (ST) as stability indicator of dough is significantly
influenced by dough constituents. As depicted, the lowest and high-
est ST are found in F16 (control) and F15 (long chain inulin-­containing
IRWF at 20% w/w) with contents equal to 8.73 and 19.23 min, re-
spectively. In other words, wheat dough supplementation using
IRWF enhanced the ST significantly on the basis of its supplemen-
tation level and polymerization degree of incorporated inulin. In this
regard, it seems that dough samples with higher IRWF incorporation
level had higher ST and consequently mixing resistance compared
to the control sample. Previously, it has been stated that differently
originated proteins in dough mixtures are not fully hydrated and
behaved as dispersed particles which reinforce the gluten network
and consequently increase their derived dough ST (Paraskevopoulou
et al., 2010).
Considering results obtained in Table 2, the farinograph quality
number (FQN) which is an indicator of gluten protein quality and
quantity is enhanced by IRWF supplementation level and inulin DP.
In other words, the highest FQN is found in F15 with about 98.46%
enhancement in FQN compared to control sample. As the FQN is
an indicator of mixing tolerance and dough strength, it is suggested
that IRWF incorporation enhances the bread-­making performance
of wheat dough (Whitney & Simsek, 2020).
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6      MOLLAKHALILI-­MEYBODI et al.

3.2  |  Effects of IRWF on physicochemical
characteristics
The impacts of IRWF supplementation on physicochemical charac-
teristics of wheat breads have been investigated and represented
in Table 3.
As depicted, specific volume (SV) is significantly influenced by
supplementation level and the type of incorporated inulin. In other
words, despite no significant difference between F16 (Control sam-
ple) and F1 (containing S inulin type and supplementation level at
10% w/w) (p  ≥ .05), about 48.30% w/w decreased is found in SV
of F15 compared to control sample (p  < .05) at. As SV is affected
by dough strength and its prospective in restoring gases through
bread-­making (Kou et al., 2019; Whitney & Simsek, 2020), this re-
duction in IRWF supplemented formulations may be induced by
dough elasticity reduction provoked by their weakening impacts on
gluten network. However, no gluten dilution is observed in all sup-
plemented formulations, it seems more than early fixing of structure
in L-­t ype inulin-­containing IRWF formulation induced by their higher
water content leads to retardation in the development of gluten net-
work (Sabanis & Tzia, 2009). In other words, higher affinity of L-­t ype
inulin to bound water resulted in its competence with gluten and
consequently reduce its development and ability to restore gases
produced through fermentation process. This finding is in accor-
dance with (de Erive et al., 2020).
The textual characteristics of IRWF-­supplemented-­wheat breads
in terms of hardness, chewiness, and springiness are demonstrated
in Table 3. As depicted, the highest and lowest hardness is found in
F16 and F15 at 756.09 and 3189.11 g, respectively. Changes in chew-
iness characteristics are also in similar order of harness. In other
words, the crumb hardness and chewiness have been increased by
IRWF supplementation level and polymerization degree of incor-
porated inulin (p < .05). Considering the impact of inulin on crumb
hardness, however, it has previously stated that its incorporation in
wheat bread provides a coarse crumb structure with higher hardness
(Frutos et al., 2008; Sardabi et al., 2021; Sirbu & Arghire, 2017), its
adverse impact has been found at this study which is improved by
increasing the inulin degree of polymerization. On the other hand,
despite decreased hardness and increased SV via gluten protein in-
clusion in steamed bread (Qian et al., 2019), a reverse order has been
found at this study which is supposed to be induced by more coarse
crumb structure formed by formulations replaced by IRWF at high
level, especially those containing long-­chain inulin as demonstrated
by (Mohammadi et al., 2021). As a less elastic system is formed
through IRWF supplementation, it is also declared by springiness
data. Springiness as an indicator of deformation ratio of a deformed
product to its un-­deformed state after removal of deforming force is
directly dependent on hardness and chewiness. Generally, any vari-
ation in formulation will be corresponded in springiness and chewi-
ness differentiation (Eriksson et al., 2014). Increasing the springiness
value denote a well-­developed gluten network with more elasticity
in crumb. As demonstrated in Table 3, IRWF incorporation reduced
the springiness value depending on its incorporation level and po-
lymerization degree of incorporated inulin. In other words, it has
been decreased by increasing the IRWF incorporation level and in-
creasing the inulin DP with springiness values equal to 9.03 and 5.82

TA B L E 3  Physicochemical
Parameters
characteristics of wheat breads
Springiness supplemented with inulin-­containing
Samples SV (cm3/g) Hardness (g) Chewiness (mJ) (mm) reconstituted wheat flour

F1 2.63 ± 0.06a 812.32 ± 2.23L 84.36 ± 0.07j 9.14 ± 0.07a

b k i
F2 2.58 ± 0.02 1009.48 ± 0.17 92.76 ± 0.01 8.94 ± 0.02ab
F3 2.53 ± 0.07bc 1319.67 ± 2.15j 115.49 ± 0.05h 8.54 ± 0.06b
c i g
F4 2.45 ± 0.05 1523.71 ± 3.08 130.09 ± 0.03 8.06 ± 0.01d
F5 2.43 ± 0.08c 1817.14 ± 1.18g 145.12 ± 0.08e 8.22 ± 0.05c
d h g
F6 2.01 ± 0.03 1620.86 ± 0.05 129.72 ± 1.00 8.03 ± 0.09d
F7 1.90 ± 0.09d 1892.31 ± 11.08g 134.17 ± 0.09f 8.68 ± 0.01b
e f e
F8 1.72 ± 0.01 2134.19 ± 10.01 142.03 ± 1.16 7.10 ± 0.08f
F9 1.68 ± 0.04e 2315.49 ± 4.12e 151.18 ± 1.02d 7.03 ± 0.05f
f d bc
F10 1.43 ± 0.03 2618.72 ± 9.60 161.12 ± 1.09 6.12 ± 0.04h
F11 1.58 ± 0.08e 2198.02 ± 4.38f 135.83 ± 0.08f 7.60 ± 0.03e
ef e e
F12 1.51 ± 0.04 2445.29 ± 0.12 147.69 ± 1.35 6.78 ± 0.05g
F13 1.49 ± 0.06f 2743.81 ± 5.12c 158.00 ± 0.02c 6.07 ± 0.01h
f b b
F14 1.45 ± 0.05 3025.13 ± 3.19 169.15 ± 1.13 5.56 ± 0.05i
F15 1.37 ± 0.01g 3189.11 ± 8.01a 183.14 ± 1.81a 5.82 ± 0.08i
a m k
F16 2.65 ± 0.08 756.09 ± 1.13 70.06 ± 0.04 9.03 ± 0.03a

Note: Data are reported as average ± standard deviation. The values with the different lowercase
letter in each column mean the significant difference (p < .05).
 |

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MOLLAKHALILI-­MEYBODI et al.       7

for control and F16 samples, respectively. The decrease observed in
springiness characteristics of IRWF-­incorporated wheat bread for-
mulations containing L-­t ype inulin could be attributed to their lower
strength and gas retention capability of gluten network. However,
incomplete starch swelling through baking will also restrict the
sponge-­like formation in bread crumb. This result is verified (de Erive
et al., 2020; Londono et al., 2015).
3.3  |  Effects of IRWF on color parameters
Color characteristics are considered to be influential in consumer's
acceptance determination of bread alongside its textural and physic-
ochemical characteristics. Two mechanisms of Maillard and caramel-
ization reactions are simultaneously evolved in color development of
bakery products like wheat bread (Anil, 2007). The crust and crumb
color determination of IRWF-­supplemented wheat bread has been
conducted by CIE L*a*b* color scale as represented in Table 4. The
crust and crumb color are generally developed by Maillard/carameli-
zation reactions and inherent color characteristics of formulation,
respectively (Dhen et al., 2018). Crumb color is also influenced by
structure forming characteristics of formulation through bread-­
making process (NithyaBalaSundari et al., 2020).
As demonstrated in Table 4, IRWF-­supplemented bread had sig-
nificantly (p < .05) lower L*, and higher a* and b* in crust compared
to the control bread. Considering the polymerization degree of inu-
lin, the highest variation is found in S-­t ype inulin samples. In other
words, the lowest L* value is found in F5 at 72.18. The lightness re-
duction via low DP inulin incorporation has been previously stated
by (Aguiar et al., 2021) which was supposed to be induced by their
more susceptibility to be involved in Millard reaction. Decreased L*
along with increased a* of wheat bread crust was previously related
to increased Millard reaction ratio by (de Erive et al., 2020; Skendi
et al., 2010). As the trend of changes in crust L* is inconsistent with
a* and b* values, it seems that the yellow-­red shade of proteins in
crust color determination is also influensive (de Erive et al., 2020).
As mentioned previously, the crumb color characteristics of bread
samples are influenced by their formulation and structure-­forming
characteristics (Kou et al., 2019). As demonstrated crumb color char-
acteristics of wheat bread is influenced by its supplementation level
and polymerization degree of incorporated inulin (p < .05). In other
words, the highest and lowest L* are found in F15 (supplemented by
20% w/w L-­t ype inulin-­containing IRWF) and F5 (supplemented by
20% w/w S-­t ype inulin-­containing IRWF) with values at 79.12 and
89.15, respectively. It seems that higher susceptibility of S-­t ype inu-
lin to be fermented by yeast leads to more release of carbon dioxide
and air bubble expanding through bread-­making process which re-
sulted in more light scattering and consequently decreased L* value.
Increased L* in F15 is accompanied by the highest b* and harness
values which are in accordance with (Matos & Rosell, 2013).
3.4  |  Sensory evaluation of IRWF supplemented
wheat bread
Considering the importance of the overall acceptability of new
products to be introduced in market, sensory assessment of IRWF-­
supplemented wheat bread was done and reported in Figure  1.

TA B L E 4  Crust and crumb color analysis of wheat breads supplemented with inulin-­containing reconstituted wheat flour

Crust Crumb

Trial L* a* b* L* a* b*
c a e c a
F1 77.28 ± 0.01 9.13 ± 0.01 24.15 ± 0.01 89.17 ± 0.05 9.17 ± 0.05 15.23 ± 0.01f
cd a e e a
F2 76.14 ± 0.07 9.95 ± 0.06 23.92 ± 0.01 85.13 ± 0.11 9.23 ± 0.01 15.94 ± 0.11f
F3 75.79 ± 0.02d 9.84 ± 0.04a 23.17 ± 0.05f 82.56 ± 0.14f 9.36 ± 0.07a 16.32 ± 0.07e
d a e g a
F4 73.13 ± 0.09 10.13 ± 0.01 25.14 ± 0.08 80.00 ± 0.08 9.63 ± 0.03 18.01 ± 0.03c
F5 72.18 ± 0.11e 10.27 ± 0.05a 27.92 ± 0.03bc 79.12 ± 0.21g 9.59 ± 0.01a 17.59 ± 0.01d
bc a f a b
F6 78.00 ± 0.21 9.15 ± 0.07 23.14 ± 0.01 91.15 ± 0.17 8.21 ± 0.05 17.21 ± 0.09d
F7 77.18 ± 0.08c 8.23 ± 0.01b 23.56 ± 0.10 f 90.83 ± 0.06a 8.00 ± 0.08b 19.14 ± 0.02b
c c e b b
F8 76.21 ± 0.08 7.16 ± 0.08 24.14 ± 0.09 90.17 ± 0.03 7.93 ± 0.02 18.91 ± 0.21c
F9 76.09 ± 0.05cd 9.11 ± 0.05a 26.11 ± 0.07d 91.51 ± 0.05a 6.82 ± 0.04c 19.18 ± 0.17b
d c a a b
F10 75.73 ± 0.17 7.69 ± 0.02 28.91 ± 0.02 91.00 ± 0.19 8.00 ± 0.01 19.02 ± 0.02b
F11 79.13 ± 0.06b 6.15 ± 0.02d 26.21 ± 0.01d 83.12 ± 0.11f 7.21 ± 0.00 c 17.00 ± 0.08d
b d c e c
F12 79.00 ± 0.05 6.32 ± 0.05 27.14 ± 0.02 85.17 ± 0.28 6.95 ± 0.03 17.23 ± 0.04d
F13 78.94 ± 0.10 b 4.18 ± 0.01f 28.00 ± 0.08b 87.99 ± 0.23d 7.63 ± 0.04b 18.62 ± 0.09c
b c b d
F14 80.01 ± 0.01 7.09 ± 0.07 28.23 ± 0.06 88.54 ± 0.04 6.12 ± 0.01 18.14 ± 0.11c
F15 81.12 ± 0.13ab 7.38 ± 0.03c 29.11 ± 0.03a 89.15 ± 0.10 c 7.79 ± 0.06b 20.34 ± 0.05a
a e g d d
F16 83.56 ± 0.11 5.88 ± 0.05 19.82 ± 0.01 87.19 ± 0.07 5.36 ± 0.05 18.83 ± 0.08c

Note: Data are reported as average standard error. Values with different lowercase letters according to Tukey's test are significantly different in each
column (p < .05).
 |
8      MOLLAKHALILI-­MEYBODI et al.
Sensory characteristics were evaluated at four criteria of texture,
flavor, color, and overall acceptability. As depicted, all samples are
higher than mediate at all evaluated criteria. Generally, the highest
score considering flavor, color, texture, and overall acceptability
has been gained by F5 (containing S-­t ype inulin at IRWF supple-
mented level of 20% w/w). As the specific volume and crust color
are the main indicators to meet the expectative of consumers (Di
Monaco et al., 2015; Pasqualone et al., 2019), the optimization is
found in F5.
Considering the flavor sensation, the highest score is found in
samples containing S-­t ype inulin which can be attributed to higher
metabolites of Millard reaction as described by (Mohammadi
et al., 2021). The overall acceptability is well matched with texture
characteristics (r = 0.87, p < .05).
3.5  |  Nutritional value
Nutritional value of wheat breads supplemented with IRWF
as Total Sugar (S), Resistant Starch (RS), Digestible Starch (DS),
Hydrolysis Index (HI), Predicted Glycemic Index (PGI), and fructan
loss is presented in Table 5. As demonstrated, TS, RS and DS con-
tents in the products are in the range of 64.13%–­70.17%, 5.03%–­
10.19%, and 57.13%–­
68.09% on dry weight basis, respectively.
Considering the control sample, incorporation of IRWF enhanced
the RS and decreased the DS depending on its supplementation
level and the polymerization degree of inulin incorporated in IRWF
formulations. The decrease in TS seems to be more pronounced at
formulations containing lower DP inulin. As the lower DP inulin is
more susceptible to be hydrolyzed through bread making process,
it will be more remarkably lost by Millard reaction through bak-
ing by Millard reaction (Morreale et al., 2019). However, this de-
crease will be reversely influenced by increasing the level of IRWF
incorporation. The increased Millard reaction ratio through S-­t ype
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F I G U R E 1  Radar plot for sensory
evaluation of wheat breads supplemented
with inulin-­containing reconstituted
wheat flour
inulin incorporation is also obvious in crust color consideration.
The highest RS value is found in F15 containing the highest IRWF
supplementation level provided by higher DP inulin with amount
at 10.19% which simultaneously has the lowest DS of 58.12%.
Despite similar inulin incorporation level at different formulations,
the difference in the rate of digestion seems to be induced by the
differences in their nature and impacts on starch gelatinization. As
the inulin gel formed at lower temperature, it prohibits the water
accessibility for the amorphous region of starch and consequently
restricts its gelatinization (Luo et al., 2017; Reshmi et al., 2017).
Protein inclusion is also reported to reduce the starch susceptibility
to enzymatic attack by decreasing the starch granule swelling and
its gelatinization (Graça et al., 2021). In this regard, increased RS
has been found in all formulations by increasing the level of IRWF.
Hydrolysis indices (HI) which is calculated by comparison of
glucose released from sample and standard glucose through the
digestion process and its corresponding pGI is considerably in-
fluenced by the supplementation process. As depicted in Table 4,
the highest and lowest HI is found at F16 and F15 with values
at 57.08% and 22.6%, respectively, with simultaneously the high-
est and lowest pGI. In other words, increasing the polymerization
degree of incorporated inulin significantly (p < .05) decreased the
HI which are also decreased more sharply at formulations con-
taining higher IRWF supplementation level. The decreased HI in-
duced by increasing the level of IRWF and polymerization degree
of incorporated inulin is attributed to decreased starch gelatini-
zation ratio and its preventative impacts on hydrolyzing enzymes
(Englyst et al., 2007; Garsetti et al., 2005; Gourineni et al., 2017).
Considering the pGI, Foods with GI lower than 55, 56–­69, and
higher than 70 are categorized as low, middle, and high GI foods,
respectively. The pGI of formulations prepared in this study ranged
from 51.93 to 71.05 with all IRWF-­supplemented formulations as
medium GI and F14 and F15 samples recognized as low GI ones
(Reshmi et al., 2017). This observation is believed to be induced by
 |

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MOLLAKHALILI-­MEYBODI et al.       9

TA B L E 5  In vitro nutritional characteristics of wheat breads supplemented with inulin containing reconstituted wheat flour

Parameters

Samples Total sugar Resistant starch HI PGI Digestible starch Fructan loss
b e b b c
F1 68.01 ± 0.02 5.32 ± 0.09 42.59 ± 0.04 63.09 ± 0.01 63.13 ± 0.07 43.01 ± 0.09a
F2 66.09 ± 0.05d 6.03 ± 0.12d 41.38 ± 0.05c 62.43 ± 0.08c 60.17 ± 0.04d 43.18 ± 0.05a
f d d d e
F3 64.13 ± 0.07 6.32 ± 0.04 40.49 ± 0.07 61.94 ± 0.11 58.91 ± 0.11 43.09 ± 0.11a
F4 64.25 ± 0.03f 7.05 ± 0.06c 39.01 ± 0.09e 61.13 ± 0.06d 57.13 ± 0.09f 41.12 ± 0.06b
e c f e a
F5 65.37 ± 0.09 7.35 ± 0.03 37.26 ± 0.00 60.17 ± 0.11 68.09 ± 0.03 41.00 ± 0.07b
F6 67.18 ± 0.01c 7.00 ± 0.07c 35.52 ± 0.12g 59.21 ± 0.09 f 60.13 ± 0.16d 39.14 ± 0.01c
c b i h e
F7 67.03 ± 0.08 8.61 ± 0.13 31.69 ± 0.10 57.11 ± 0.06 59.21 ± 0.07 38.54 ± 0.09d
F8 67.92 ± 0.04b 9.07 ± 0.08b 31.38 ± 0.04i 56.94 ± 0.04h 58.03 ± 0.12f 38.00 ± 0.03d
b b i h f
F9 67.89 ± 0.07 9.21 ± 0.05 31.51 ± 0.09 57.01 ± 0.08 58.11 ± 0.09 37.19 ± 0.04e
F10 68.11 ± 0.04b 7.51 ± 0.02 30.01 ± 0.17j 56.19 ± 0.02i 61.03 ± 0.04d 36.71 ± 0.05f
b b h g e
F11 68.00 ± 0.02 8.83 ± 0.03 34.01 ± 0.05 58.38 ± 0.07 59.78 ± 0.06 30.12 ± 0.09g
F12 68.02 ± 0.04b 9.05 ± 0.12b 29.76 ± 0.11j 56.05 ± 0.05i 59.81 ± 0.09e 29.18 ± 0.13h
b a k j e
F13 68.31 ± 0.03 9.89 ± 0.07 26.96 ± 0.01 54.51 ± 0.01 59.00 ± 0.12 28.64 ± 0.04hi
F14 69.51 ± 0.04a 10.01 ± 0.04a 24.34 ± 0.08L 53.07 ± 0.09k 58.01 ± 0.05f 27.13 ± 0.07i
a a m k f
F15 69.87 ± 0.08 10.19 ± 0.15 22.26 ± 0.05 51.93 ± 0.1 58.12 ± 0.09 25.42 ± 0.05j
F16 70.17 ± 0.07a 5.03 ± 0.02e 57.08 ± 0.03a 71.05 ± 0.05a 65.14 ± 0.07 b -­

Note: Data are reported as average standard error. Values with different lowercase letters according to Tukey's test are significantly different in each
column (p < .05).

increasing the level of RS and inulin and their inhibitory impacts
on enzymes participated in post-­p randial glucose response. GI is
a nutritional quality determinative parameter and diets with low
GI and high RS are believed to be therapeutic in diabetic patients,
reducing the insulin resistance and blood glucose adjustment, im-
proving the lipid metabolism, and preventing the cardiovascular
diseases (Chung et al., 2011; Reshmi et al., 2017).
Inulin is incorporated as a functional ingredient in this study
through its prebiotic role. Consequently, its stability through bread
making process is considerably important, as influensive prebiotics
needs to be entered intact to upper intestinal tract. As the incor-
poration level of inulin was different, monitoring the fructan loss
in IRWF-­
supplemented formulations indicate a progressive fruc-
tan loss through processing in the range of 25.42%–­43.01% w/w.
In other words, the highest and lowest fructan loss is found in F15
and F1, respectively, which indicate the enhanced prebiotic loss by
decreasing its incorporation level and polymerization degree. The
enhanced inulin loss by decreasing its polymerization degree is at-
tributed to its higher susceptibility to be used by yeast and take part
in Millard reaction. This finding is in accordance with (Mohammadi
et al., 2021).
4  |   CO N C LU S I O N
gluten dilution impacts, adverse impacts on technological charac-
teristics are still restricting in industrially production of them espe-
cially long-­chain inulin-­containing ones which are attributed to their
higher water absorption content, accelerated gluten network forma-
tion, and decreased starch gelatinization. However, long-­chain inulin
incorporation significantly improves the nutritional characteristics
of wheat considering resistant starch content, hydrolysis index, pre-
dicted Glycemic index, and fructan loss parameters. Consequently,
whole bread-­
making process as the type of fermentation, time:
temperature ratio of fermentation process and increasing ratio of
temperature through baking process should be optimized to formu-
late optimized formulation from both nutritional and technological
perspective.
AC K N OW L E D G M E N T
The financial support of this study is achieved by the Shahid
Sadoughi University of medical science, Yazd, Iran.
C O N FL I C T O F I N T E R E S T
None declared.
DATA AVA I L A B I L I T Y S TAT E M E N T
The authors confirm the availability of all data supporting the find-
ings within the article.

Inulin supplementation of wheat bread in the form of inulin recon- E T H I C S S TAT E M E N T

stituted wheat flour (IRWF) seems to be applicable from nutritional The Ethical aspects of this study are evaluated and approved by the
perspective, especially at formulations containing long-­chain inulin Institutional Review Board of Shahid Sadoughi University of Medical
type. However, despite gluten inclusion at formulations to conquer Sciences.
 |
10      MOLLAKHALILI-­MEYBODI et al.
ORCID
Neda Mollakhalili-­meybodi  https://orcid.
org/0000-0001-8473-7517
Bahador Hajimohammadi  https://orcid.
org/0000-0003-4463-5465
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How to cite this article: Mollakhalili-­meybodi, N.,
Ehrampoush, M. H., Hajimohammadi, B., & Mosaddegh, M. H.
(2022). Formulation optimization of functional wheat bread
with low glycemic index from technological and nutritional
perspective. Food Science & Nutrition, 00, 1–11. https://doi.
org/10.1002/fsn3.3060