LWT - Food Science and Technology 160 (2022) 113301

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LWT
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The influence of fermented buckwheat, quinoa and amaranth flour on

gluten-free bread quality
Saliha Yeşil a, Hacer Levent b, *
a
Karamanoğlu Mehmetbey University, Faculty of Engineering, Department of Food Engineering, 70100, Karaman, Turkey
b
Karamanoğlu Mehmetbey University, Faculty of Health Sciences, Department of Nutrition and Dietetics, 70100, Karaman, Turkey

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

Keywords: In this study, buckwheat, quinoa and amaranth flours were fermented by two different methods (spontaneous
Buckwheat fermentation and yeast fermentation) and fermented dough of pseudocereals (PFD) was used (0, 15, 30 and 45%)
Quinoa in gluten-free bread formulation. The effects of PFD and fermentation methods of pseudocereals on bread quality
Amaranth
were investigated. The increase in the PFD ratio enhanced the amounts of ash, crude protein, total phenolic
Bread
Fermentation
content (TPC) and antioxidant activity (AA) of breads samples. Among the pseudocereals, quinoa revealed the
highest TPC and AA values in the bread samples. The high ratio of PFD (45%) increased the Ca, P, K, Fe, Mg and
Zn content of gluten-free breads by 1.3–4.3 times. Although breads with fermented dough of amaranth received
high scores in terms of symmetry, pore structure and appearance, they were less appreciated by consumers in
terms of taste-odor. Fermentation of pseudocereals using bakers’ yeast yielded in softer crumb in the bread
samples. Breads containing PFD produced by spontaneous fermentation scored less in sensory analyses. It was
concluded that acceptable quality gluten-free bread can be produced with the use of up to 30% PFD.

1. Introduction than their gluten-containing counterparts (Kupper, 2005; Fasano &

Celiac disease (CD) is a chronic inflammatory disorder of small in­
testine triggered by the intake of gluten proteins present in wheat, rye
and barley in genetically susceptible individuals (Fasano & Catassi,
2012). Ingestion of gluten in patients with CD leads to an enteropathy
with disruption of the mucosal surface followed by abnormal absorption
of nutrients (Parzanese et al., 2017). CD has emerged as an important
public health problem in the last 20 years (Singh et al., 2018). CD is
associated with various diseases such as enamel hypoplasia, dermatitis
herpetiformis, iron deficiency anemia, short stature, osteoporosis,
recurrent oral aphthae, various neurological and endocrine diseases,
arthritis, various types of cancer, female infertility, persistent chronic
hypertransmia-nasaemia of unknown origin, and other autoimmune
disorders (Guandalini & Assiri, 2014; Rodrigo, 2006).
For CD patients, the only currently available and effective treatment
is a strict, lifelong gluten-free diet (Ludvigsson & Murray, 2019). In
addition to the mandatory gluten-free diet due to CD, the consumption
of gluten-free products has increased significantly in recent years. Un­
fortunately, gluten-free foods contain less B vitamins, iron, zinc, mag­
nesium, D vitamin and dietary fiber and have lower nutritional quality
Catassi, 2012). Results from studies show that celiac patients have
insufficient intake of some essential nutrients, as well as an unbalanced
intake of carbohydrates, fats and proteins, compared to controls
(Alvarez-Jubete, Arendt, & Gallagher, 2010). Therefore, gluten-free diet
may cause some health problems due to inadequate and unbalanced
nutrition.
The pseudocereals buckwheat, quinoa and amaranth have received a
lot of attention in recent years due to their excellent nutrient profiles and
potential health benefits (Alvarez-Jubete, Wijngaard, Arendt, & Gal­
lagher, 2010). Pseudocereals contain high protein with balanced amino
acid composition, and are also an important source of phytochemicals,
dietary fibers, vitamins and minerals (Yaver & Bilgiçli, 2020). In addi­
tion, buckwheat, quinoa and amaranth seeds are naturally gluten-free
and are considered as healthy ingredients for fortifying gluten-free
foods (Kupper, 2005; Alvarez-Jubete, Wijngaard, et al., 2010).
Gluten-free breads, which are generally produced from gluten-free
flours and starches that lack the gluten proteins, have poor sensory,
textural properties and nutritional values (Ua-Arak, Jakob, & Vogel,
2017). Buckwheat, quinoa and amaranth have been used in many
studies in the literature to ensure adequate nutritional intake in

* Corresponding author. Department of Nutrition and Dietetics, Karamanoğlu Mehmetbey University, Ibrahim Oktem Street, 70100, Karaman, Turkey.
E-mail addresses: salihayesil12@gmail.com (S. Yeşil), hacerlevent@hotmail.com (H. Levent).

https://doi.org/10.1016/j.lwt.2022.113301
Received 15 September 2021; Received in revised form 27 January 2022; Accepted 26 February 2022
Available online 7 March 2022
0023-6438/© 2022 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
S. Yeşil and H. Levent LWT 160 (2022) 113301

individuals with CD (Alvarez-Jubete, Wijngaard, et al., 2010; Chlopicka, Table 1

Pasko, Gorinstein, Jedryas, & Zagrodzki, 2012; Demir & Bilgiçli, 2021). Gluten-free bread ingredients.
In a study conducted by Bilgiçli & İbanoğlu (2015), buckwheat and Ingredients Control Enriched breads
quinoa flour replaced wheat flour up to 20% in the bread formulation
15% 30% 45%
and it was reported that Fe, K, Mg, Mn, and Zn contents of the breads
increased with increasing levels of quinoa and buckwheat flour. Chlo­ GF flour mix (RF/CS, 50:50) (g) 100 85 70 55
Fresh bakers’ yeast (g) 3 3 3 3
picka et al. (2012) reported that the addition of buckwheat flour (15% Salt (g) 1.5 1.5 1.5 1.5
and 30%) to bread improves the antioxidant properties of bread, as well Sugar (g) 6 6 6 6
as its sensory properties. Baking powder (g) 2 2 2 2
There are several studies in the literature that gluten-free breads Whole milk powder (g) 5 5 5 5
Sunflower oil (ml) 5 5 5 5
produced with gluten-free flours and/or starches are not only nutri­
Guar gum (g) 1 1 1 1
tionally but also very poor in terms of sensory and textural aspects. It is PFD 0 15 30 45
reported that it is difficult for consumers to find gluten-free breads with Water (ml) 180 150–170 130–160 120–150
a desirable appearance, texture, taste and mouthfeel properties (Capriles
GF: Gluten-free, RF: Rice flour, CS: Corn starch, PFD:Fermented dough of
& Arêas, 2014; Foschia, Horstmann, Arendt, & Zannini, 2016; Gal­ pseudocereals.
lagher, Gormley, & Arendt, 2004; Moroni, Dal Bello, & Arendt, 2009).
Nowadays, due to consumer demand for clean labels, the importance
(2007). In the spontaneous fermentation method, 420 g of pseudocereal
of technologies that allow high quality gluten-free breads without the
flour was mixed with 670 g of tap water and the mixture was fermented
need for additives has increased (Bender & Schönlechner, 2020).
at 30 ◦ C for 20 h. In sourdoughs prepared using bakers’ yeast, 4.2 g of
Fermentation can be defined as a desirable biochemical modification
fresh bakers’ yeast was added to the mixture and fermented at the same
process of the primary food matrix produced by microorganisms and
temperature and time.
their enzymes (Karovičová & Kohajdova, 2007). Fermentation of cereals
or sourdough fermentation positively improves the taste and aroma,
2.2.2. Gluten-free bread preparation
texture, nutritional value and shelf-life properties of bakery products
Ingredients of gluten-free bread samples are given in Table 1. For
(Carnevali, Ciati, Leporati, & Paese, 2007; Gobbetti, Rizzello, Di Cagno,
control gluten-free bread (0% PFD) production, 100 g gluten-free flour
& De Angelis, 2014). The development of fermentation technology for
mix (rice flour/corn starch, 50:50), 3 g fresh bakers’s yeast, 1.5 g salt, 6 g
flour alternatives to wheat can be considered as an important opportu­
sugar, 2 g baking powder, 5 g whole milk powder, 5 ml sunflower oil, 1 g
nity to meet the demand for more natural and healthy food (Coda, Di
guar gum and 180 ml water were used. The amount of water used in
Cagno, Gobbetti, & Rizzello, 2014). Bakers’ yeast, which is used for
making bread was determined by preliminary experiments according to
bread fermentation all over the world, is very important for bread
dough properties. The bread ingredients were mixed to a fluid homo­
quality (Birch, Petersen, Arneborg, & Hansen, 2013). Yeast plays a vital
geneous dough consistency in laboratory type mixer (Kitchen-aid,
role in bread production due to the production of CO2, ethanol, organic
Artisan Series, Greenville, OH, USA) for 5 min, followed by mass
acids, glycerol and aroma compounds and other metabolites that have
fermentation at 30 ◦ C at 85% relative humidity for 30 min. The fer­
an impact on dough rheology, bread texture, volume and taste (Struyf
mented dough was baked at 175 ◦ C for 25 min (Bosch HGD52D120T,
et al., 2017).
Istanbul, Turkey). For pseudocereal enriched breads, the fermented
To the best of our knowledge, the use of buckwheat, quinoa and
dough of buckwheat, quinoa and amaranth replaced the gluten-free
amaranth sourdoughs produced by spontaneous and yeast fermentation
flour mix in the formulation. The bread experiments were carried out
to improve the chemical, textural and sensory quality of gluten-free
by applying a 3 (pseudocereal type) × 2 (fermentation method) × 4(PFD
bread did not investigated. Therefore, the aims of this study were (1)
ratio) × 2 factorial design. The gluten-free bread production was per­
to enrich gluten-free breads with pseudocereals to meet the nutritional
formed in duplicate and mean values were reported. After baking, the
needs of individuals following a gluten-free diet, (2) to improve the
breads were allowed to cool for 1 h and then packed in polyethylene
sensory and physical properties of gluten-free breads by fermentation as
bags.
well as nutritional enrichment with pseudocereals, and (3) to determine
the impacts of pseudocereal type, fermentation method and pseudo­
2.2.3. Chemical analyses
cereal fermented dough (PFD) ratio on gluten-free bread quality.
AACC methods were used to determine the moisture (method
44–19), ash (method 08–01), crude protein (method 46–12) and crude
2. Materials and methods
fat (method 30–25) content of rice flour, corn starch, buckwheat,
quinoa, amaranth and bread samples (AACC, 1990). Phytic acid was
2.1. Materials
measured colorimetrically as described by Haug and Lantzsch (1983).
For phytic acid analysis, the samples were extracted with hydrochloric
Rice flour, corn starch, bakers’ yeast, salt, granulated sugar, baking
acid solution, and the phytic acid in the sample was precipitated with Fe
powder and sunflower oil were procured from local markets in Karaman,
III solution. The amount of iron in the extract was determined spectro­
Turkey. Whole milk powder was supplied from Ova Dairy Products Inc.,
photometrically. The amount of phytic acid was calculated and given as
Konya, Turkey. Buckwheat, quinoa and amaranth were procured from
mg/100 g.
Yayla Agro Food Industry & Transport Inc., Mersin, Turkey. Guar gum
Total phenolic content (TPC) was determined colorimetrically using
was obtained from Kimbiotek Chemical Substances Inc., Istanbul,
the Folin-Ciocalteu reagent. Ground samples (1 g) were extracted in 10
Turkey. Pseudocereals were ground with a lab scale mill (Bosch,
ml of solvent (methanol/hydrochloric acid/distilled water, 8:1:1, v/v/v)
MKM600) and sieved passed through a 500 μm sieve to obtain whole
for 2 h at room temperature. Then, the extract was centrifuged at 3000
flour.
rpm for 10 min (Beta, Nam, Dexter, & Sapirstein, 2005; Gao, Wang,
Oomah, & Mazza, 2002). The extract (0.1 ml), Folin-ciocalteu reagent
2.2. Methods
(0.5 ml, 10%, v/v, water) and sodium carbonate solution (1.5 ml, 20%,
w/v) were mixed in the test tube and filled to 10 ml with distilled water.
2.2.1. Sourdough preparation
After mixing the contents of the tube, it was kept at room temperature
Sourdoughs of buckwheat, quinoa and amaranth flours were pre­
and dark place for 2 h and absorbance values were measured at 760 nm
pared in 2 different ways (spontaneous fermentation and yeast
with spectrophotometer (Shimadzu UV1800, Japan). TPC was expressed
fermentation) according to the method suggested by Katina et al.

2
S. Yeşil and H. Levent LWT 160 (2022) 113301

Table 2 determined. Ground sample (1 g) was mixed with of pure nitric acid (15
Chemical properties of raw materials.a. ml) in a burning cup and kept at room temperature for 1 night. Next, 4
Rice Corn Buckwheat Quinoa Amaranth ml of perchloric acid was added and the samples were gently heated for
flour starch 5–6 h. After the samples were cooled to room temperature, 5 ml of
Moisture (g/ 11.5 ± 8.5 ± 9.4 ± 0.14b 8.6 ± 7.7 ± hydrogen peroxide was added and reheated until discoloration was
100 g) 0.18a 0.21c 0.14c 0.28d observed. Mineral contents of filtrates were determined by the ICP-OES
Ash (g/100 0.90 ± 0.10 ± 2.24 ± 2.02 ± 2.73 ± (Inductively Coupled Plasma Optical Emission Spectrometry, Agilent
g) 0.30c 0.03d 0.07a 0.05b 0.29a 720, USA) (Levent, Koyuncu, Bilgiçli, Adıgüzel, & Dedeoğlu, 2020).
Crude 9.50 ± 0.61 ± 12.73 ± 14.93 ± 15.03 ±
protein (g/ 0.75c 0.01d 0.61b 0.88a 0.12a
100 g) 2.2.4. Hardness of gluten-free breads
Crude fat (g/ 0.80 ± 0.43 ± 3.98 ± 6.07 ± 5.79 ± Hardness values of bread samples were measured with texture
100 g) 0.15c 0.08c 0.18b 0.22a 0.12a analyzer TA-XT plus (Stable Micro Systems, Surrey, UK) equipped with a
TPC (mg 1.39 ± 1.17 ± 4.98 ± 5.22 ± 2.55 ±
5 kg load cell. Hardness measurements were made on the middle slices
GAE/g)b 0.13c 0.08c 0.11a 0.17a 0.20b
AA 3.52 ± 2.80 ± 53.08 ± 60 .14 ± 18.46 ± of bread with a width of 25 mm, which were kept at room temperature
(Inhibition 1.02d 0.65d 0.82b 0.76a 0.93c for 24 h. An aluminum 36-mm diameter cylindrical probe was used in
%)c analysis. The sample was compressed to 25% of its original height and
Phytic acid 329.40 87.64 ± 1461.56 ± 1293.59 1748.79 ± hardness was determined as the maximum force during compression.
(mg/100 ± 6.51d 9.86e 9.3b ± 11.6c 7.48a
g)
The optimal test conditions in this study were: Trigger force 5 g, the
Ca (mg/100 5.97 ± 2.71 ± 28.04 ± 36.81 ± 157.54 ± pretest, test, and posttest speeds were 1.0 mm/s, 5.0 mm/s, and 5 mm/s,
g) 0.37d 0.21e 0.98c 0.57b 0.94a respectively.
P (mg/100 g) 261.05 151.63 432.10 ± 334.94 ± 395.00 ±
± 4.84d ± 3.5e 6.19a 4.62c 3.25b
2.2.5. Sensory analyses
K (mg/100 116.24 9.32 ± 438.10 ± 641.96 ± 459.21 ±
g) ± 6.22d 1.36e 3.54c 4.59a 4.04b The sensory characteristics of gluten-free bread samples were eval­
Fe (mg/100 0.46 ± 0.21 ± 2.26 ± 3.28 ± 6.36 ± uated for symmetry, pore structure, taste-odor, appearance, chewiness,
g) 0.06e 0.08d 0.04c 0.05b 0.06a and overall acceptability by 15 semi-trained panelists (8 females and 7
Mg (mg/100 36.12 ± 2.07 ± 228.77 ± 199.55 ± 237.69 ± males, aged 22 to 48) from Karamanoğlu Mehmetbey University
g) 2.02d 1.26e 2.74b 3.43c 2.76a
Department of Food Engineering and Nutrition and Dietetics. Panelists
Zn (mg/100 0.97 ± 0.28 ± 2.51 ± 3.03 ± 2.93 ±
g) 0.13c 0.11d 0.20b 0.16a 0.22ab were selected from regular bread consumers according to the criteria of
a
being healthy, not smoking, not allergic to gluten/wheat products.
Means followed by the different letter within a raw are significantl’y
Sensory analyzes of breads were made on the same day of baking. Breads
different (P < 0.05). Values are the average of triplicate measurements on the
were served to the panelists on plastic plates at ambient temperature.
duplicate samples. Chemical properties except moisture are based on dry matter.
b
TPC: Total phenolic content. Between assessments, water was provided for mouth rinse. Panelists
c
AA: Antioxidant activity. were asked to rate the bread samples using a 1–7 scale (1: dislike very
much, 7: like very much) (Epler, Iv, & Kemp, 1998).
as gallic acid equivalent (GAE) per g of sample on dry matter basis.
The antioxidant activities (AA) of the samples were determined ac­ 2.2.6. Statistical analyses
cording to the DPPH (2-2-diphenyl-2-picrylhydrazyl) method (Beta All analyses were the average of triplicate measurements on the
et al., 2005; Gyamfi, Yonamine, & Aniya, 1999). For analysis, samples duplicate samples. For statistical analysis, SPSS statistical program
were extracted as in total phenolic analysis and treated with DPPH. (Version 22.0. Armonk, NY: IBM Corp.) was used. The data were ana­
Absorbance was measured at 517 nm at 0 and 30 min. Percentage of lysed for its statistical significance using Duncan multiple comparision
inhibition was calculated according to the formula below. test. Results are given as mean ± standard deviation and the averages
were compared at p < 0.05 level.
Inhibition %= (1 – [A of samplet = 30/A of controlt = 0]) × 100

Mineral contents of raw material and bread samples were

Table 3
Chemical properties of gluten-free bread samples.a.
n Moisture (g/100 Ash (g/100 g) Crude protein (g/100 Crude fat (g/100 g) Phytic acid (mg/100 TPCb (mg GAE/ AA (Inhibition
g) g) g) g) %)c

Pseudocereal type
Buckwheat 16 51.27 ± 4.19b 3.48 ± 0.36a 6.18 ± 0.63b 3.24 ± 0.51b 196.12 ± 78.62b 1.70 ± 0.24b 9.55 ± 4.50b
Quinoa 16 53.99 ± 3.38a 3.50 ± 0.37a 6.69 ± 1.04a 3.77 ± 0.64a 190.38 ± 72.93b 1.99 ± 0.35a 10.95 ± 5.18a
Amaranth 16 53.78 ± 3.58a 3.55 ± 0.44a 6.96 ± 1.09a 3.76 ± 0.81a 210.67 ± 87.01a 1.64 ± 0.18b 6.68 ± 2.45c

Fermentation method
Spontaneous 24 53.36 ± 3.82a 3.51 ± 0.41a 6.62 ± 1.00a 3.62 ± 0.82a 200.66 ± 80.08a 1.77 ± 0.30a 9.12 ± 4.46a
Bakers’ yeast 24 52.65 ± 3.95a 3.51 ± 0.37a 6.60 ± 0.98a 3.56 ± 0.56a 197.45 ± 78.56a 1.78 ± 0.32a 9.01 ± 4.62a

PFD(%) ratiod
0 12 51.49 ± 1.22c 3.05 ± 0.03d 5.63 ± 0.24d 3.24 ± 0.05b 78.41 ± 4.77d 1.46 ± 0.09d 4.46 ± 0.41d
15 12 57.60 ± 2.35a 3.33 ± 0.14c 6.18 ± 0.43c 3.33 ± 0.53b 195.39 ± 13.99c 1.70 ± 0.25c 6.81 ± 1.84c
30 12 53.20 ± 2.77b 3.62 ± 0.07b 7.04 ± 0.70b 3.45 ± 0.60b 236.37 ± 13.28b 1.88 ± 0.23b 10.40 ± 2.40b
45 12 49.75 ± 3.44d 4.04 ± 0.14a 7.58 ± 0.96a 4.35 ± 0.75a 286.05 ± 16.06a 2.06 ± 0.24a 14.59 ± 3.36a
a
Means followed by the different letter within a column are significantl’y different (P < 0.05). Values are the average of triplicate measurements on the duplicate
samples. Chemical properties except moisture are based on dry matter.
b
TPC: Total phenolic content.
c
AA: Antioxidant activity.
d
PFD: Pseudocereal fermented dough.

3
S. Yeşil and H. Levent LWT 160 (2022) 113301

Table 4
Mineral contents of gluten-free bread samples.a.
n Ca P K Fe Mg Zn

Pseudocereal type
Buckwheat 16 54.54 ± 4.88c 384.58 ± 51.69a 231.73 ± 53.23b 0.67 ± 0.18c 35.19 ± 16.11a 0.81 ± 0.25b
Quinoa 16 55.95 ± 5.63b 360.54 ± 29.97b 260.64 ± 77.07a 0.85 ± 0.28b 35.56 ± 16.88a 0.97 ± 0.35a
Amaranth 16 75.73 ± 21.21a 379.82 ± 41.23a 228.39 ± 50.76b 1.29 ± 0.56a 35.86 ± 16.76a 0.96 ± 0.32a

Fermentation method
Spontaneous 24 61.98 ± 15.97a 375.54 ± 42.29a 239.21 ± 63.97a 0.936 ± 0.45a 35.230 ± 16.20a 0.914 ± 0.31a
Bakers’ yeast 24 62.16 ± 16.42a 374.43 ± 43.28a 241.30 ± 61.20a 0.939 ± 0.46a 35.840 ± 16.61a 0.912 ± 0.32a

PFD(%) ratiob
0 16 48.46 ± 1.54d 326.33 ± 16.56d 162.33 ± 10.25d 0.53 ± 0.12d 13.17 ± 1.26d 0.52 ± 0.12d
15 16 57.77 ± 6.55c 351.48 ± 8.13c 213.85 ± 12.47c 0.75 ± 0.18c 29.98 ± 1.53c 0.84 ± 0.14c
30 16 66.07 ± 13.16b 395.17 ± 18.09b 266.64 ± 20.40b 1.08 ± 0.36b 42.32 ± 1.51b 1.04 ± 0.14b
45 16 75.97 ± 20.37a 426.95 ± 20.37a 318.19 ± 30.71a 1.39 ± 0.50a 56.67 ± 1.69a 1.26 ± 0.19a
a
Means followed by the different letter within a column are significantl’y different (P < 0.05). Values are the average of triplicate measurements on the duplicate
samples, and are based on dry matter.
b
PFD: Pseudocereal fermented dough.

3. Results and discussion
3.1. Chemical properties of raw materials
Chemical properties of raw materials used in bread samples are given
in Table 2. Buckwheat, quinoa and amaranth have higher ash, crude
protein, fat, TPC and AA compared to rice flour and corn starch. Among
the pseudocereals, quinoa was found remarkable with its high TPC
(5.22 mg GAE/g) and AA (60.14%). Phytic acid content of raw materials
varied between 87.64 and 1748.79 mg/100 g and amaranth was the raw
material in which the highest phytic acid content was obtained. Alvar­
ez-Jubete, Arendt, and Gallagher (2009) reported the ash, protein, and
fat values of buckwheat and amaranth as 2.1%, 12.5%, 2.1% and 2.8%,
16.5%, 5.7%, respectively. Turkut, Cakmak, Kumcuoglu, and Tavman
(2016) determined the moisture, ash and protein content of quinoa flour
as 9.13%, 2.35%, 15.54%, while the same properties of buckwheat were
reported as 9.95%, 2.21% and 14.72%. Demir and Bilgiçli (2021) re­
ported the phytic acid, TPC and AA of quinoa flour as 971 mg/100 g, 1.5
mg GAE/g and 35.5%, respectively.
Ca, P, K, Fe, Mg and Zn contents of buckwheat, quinoa and amaranth
were significantly (p < 0.05) higher than rice flour and corn starch.
Buckwheat, quinoa, and amaranth had 4.8, 6.2 and 26.4 times more Ca,
3.8, 5.5, and 4.0 times more K than rice flour, respectively. When
compared with corn starch, it was determined that buckwheat, quinoa
and amaranth had 10.3, 13.6 and 58.1 times more Ca, 47.0, 68.9 and
49.3 times more K, respectively. Öncel and Demir (2019) reported that
Ca, K, Fe, Mg and Zn content of buckwheat, quinoa and amaranth flour
were 34.76, 406.08, 2.44, 195.81 and 2.66 mg/100 g (buckwheat),
48.99, 672.79, 4.27, 195.26 and 4.67 mg/100 g (quinoa) and 164.405,
458.91, 7.02, 246.82 and 5.19 mg/100 g (amaranth), respectively. In
another study, Ca, P, K, Fe, Mg and Zn contents of buckwheat, quinoa
and amaranth flour were reported as 17.5, 414, 510, 2.88, 240 and 1.94
mg/100 g (buckwheat), 77.6, 436, 559, 4.29, 196 and 2.97 mg/100 g
(quinoa) and 200, 663, 552, 7.35, 328 ve 4.55 mg/100 g (amaranth),
respectively (Mota et al., 2016).
3.2. Chemical properties of gluten-free bread samples
Chemical properties of gluten-free bread samples are presented in
Table 3. In general, it was determined that the moisture content of
breads containing PFD was higher than the control bread. The use of
pseudocereals whole flour in the preparation of PFD and the increase in
dough water absorption due to dietary fibers may have been effective on
this result. The ash contents of breads were not significantly different
from each other according to the pseudocereal type and fermentation
method. As the ratio of PFD in the formulation increased, the ash content
of the control breads increased from 3.05 g/100g to 4.04 g/100g (at
45%). Buckwheat (2.24 g/100g), quinoa (2.02 g/100g) and amaranth
(2.73 g/100g) had higher ash content compared to rice flour (0.90 g/
100g) and corn starch (0.10 g/100g) (Table 2). The high ash content of
pseudocereals was also reflected in the final product. It was determined
that the crude protein and crude fat contents of breads containing fer­
mented dough of quinoa and amaranth flour were higher than the
breads produced using fermented dough of buckwheat flour. At the 45%
enrichment ratio, the crude protein and fat content of the gluten-free
control bread increased from 5.63 g/100g and 3.24 g/100g to 7.58 g/
100g and 4.35 g/100g, respectively. Similar observations were reported
by Levent and Bilgiçli (2011) and Bhatt, Kumari, Abhishek, and Gupta
(2020) in buckwheat (up to 20%) and amaranth (up to 50%) supple­
mented cakes and muffins, respectively. It was reported that bread
samples containing quinoa and amaranth presented more lipid, protein
and ash, thus improving the nutritional value (Alencar, Steel, Alvim, de
Morais, & Bolini, 2015). In quinoa and amaranth seeds, the percentage
of bran fraction consisting of seed coat and germ is higher than in
common cereals such as corn and wheat, which explains the higher
protein and fat content of these seeds (Bressani, 2003; Alvarez-Jubete,
Arendt, & Gallagher, 2010).
The phytic acid contents of the gluten-free bread samples ranged
from 190.38 mg/100 g to 210.67 mg/100 g depending on the pseudo­
cereal type. Breads formulated with fermented dough of amaranth flour
had higher phytic acid content than breads containing buckwheat and
quinoa. Amarant, which has the highest amount of phytic acid among
the raw materials, increased the phytic acid the most in the bread
samples (Table 2). Phytic acid content of gluten-free breads increased
significantly (p < 0.05) with the increase in PFD ratio in bread formu­
lation. Although there was a decrease in the phytic acid content of
breads containing pseudocereals fermented using bakers’ yeast,
compared to spontaneous fermentation, this decrease was not significant
(p > 0.05). Similarly, Çevik and Ertaş (2019) reported that the phytic
acid content of tarhana samples containing quinoa and buckwheat flour
(up to 40%) increased with the increasing amount of buckwheat and
quinoa flour in the formulation. Phytic acid is considered as an anti­
nutrient and decreases the bioavailability of minerals. With the use of
PFD in gluten-free bread formulation, a significant increase was
observed in the amounts of all investigated minerals, and it can be
thought that this partially reduced the negative effect of phytic acid in
bread samples.
According to pseudocereal type, the highest TPC and AA values were
determined in breads containing fermented dough of quinoa, followed
by breads with fermented dough of buckwheat and amaranth. However,
Chlopicka et al. (2012) found that buckwheat improved the antioxidant
status of bread more effectively than amaranth and quinoa. The TPC and
AA of the bread samples significantly increased (p < 0.05) with the in­
crease of the PFD ratio in the formulation. Alvarez-Jubete, Wijngaard,

4
S. Yeşil and H. Levent
Fig. 1. Hardness values of gluten-free breads samples according to a) fermentation methods and b) to fermented dough of pseudocereals (PFD) ratio ( : Buckwheat,
: Quinoa, : Amaranth).
et al. (2010) reported that the TPC of gluten-free breads prepared using
buckwheat, quinoa and amaranth flour was higher than the control
bread, and the lowest TPC was obtained in breads containing amaranth
flour. In the study carried out by Wronkowska, Zielinska,
Szawara-Nowak, Troszynska, and Soral-Smietana (2010), it was re­
ported that the antioxidant activity value of gluten-free breads produced
with buckwheat flour (up to 40%) increased with increasing buckwheat
ratio. There was no significant difference in TPC and AA values ac­
cording to the fermentation method. Pseudocereals are rich in phenolic
acids and flavonoids and therefore have relatively high antioxidant ac­
tivity. Bioactive compounds in pseudocereals increase the antioxidant
activity of bakery products where it is used (Alvarez-Jubete, Wijngaard,
et al., 2010; Vollmannova et al., 2013).
5
LWT 160 (2022) 113301
3.3. Mineral composition of gluten-free bread samples
Mineral contents of bread samples are reported in Table 4. Breads
containing fermented dough of amaranth were richer in Ca and Fe
content compared to breads containing fermented dough of buckwheat
and quinoa. K content was higher in breads containing fermented dough
of quinoa whereas Zn content was higher in breads containing quinoa
and amaranth fermented doughs. When the raw material mineral anal­
ysis results were examined, high Ca and Fe content of amaranth, K
content of quinoa, and Zn content of quinoa and amaranth were found
remarkable (Table 2). A significant increase (p < 0.05) was observed in
the amount of all mineral substances examined with the increasing PFD
ratio in breads compared to the gluten-free control bread. Compared the
control, usage of PFD at maximum enrichment ratio (45%) increased Ca,
P, K, Fe, Mg and Zn content by 1.6-, 1.3-, 2.0-, 2.6-, 4.3-, and 2.4-fold,
respectively. Since most of the minerals in pseudocereal grains are
S. Yeşil and H. Levent LWT 160 (2022) 113301

Fig. 2. Sensory analyses results of gluten-free breads samples according to a) pseudocereal type b) to fermentation method and c) to fermented dough of pseu­
docereals (PFD) ratio ( :Symmetry, :Pore structure, :Taste-odor, : Appearance, :Chewiness, :Overall acceptability).

located in the bran, whole pseudocereals are good sources of minerals Fe and Zn contents of bread samples increased with increasing amaranth
(Pongrac et al., 2013; Martínez-Villaluenga, Peñas, & Hernández-Le­ ratio (0, 10, 20, 30 and 40%) in the formulation. Çevik and Ertaş (2019)
desma, 2020). In the study conducted by Sanz-Penella, Wronkowska, used buckwheat, quinoa and lupine flour in tarhana production (up to
Soral-Smietana, and Haros (2013), it was determined that Ca, K, P, Mg, 40%) and reported that the Ca, P, K, Mg, Fe and Zn content of the final

6
S. Yeşil and H. Levent
product increased with increasing buckwheat and quinoa flour ratio.
3.4. Hardness of gluten-free bread samples
The hardness values of gluten-free breads are given in Fig. 1. It was
determined that the hardness value of the breads containing PFD pre­
pared by spontaneous fermentation method was higher than the breads
containing PFD prepared by yeast fermentation. The volume of breads
containing PFD produced by spontaneous fermentation was lower than
the volume of breads produced by yeast fermentation (data not shown).
This result may have been effective on the hardness values of the bread
samples. The hardness value increased as the PFD ratio increased in all
bread types (Fig. 1.). Similar results were reported by Sanz-Penella et al.
(2013), Bilgiçli & İbanoğlu (2015) and Azizi, Azizi, Moogouei, and
Rajaei (2020). The increase in crumb hardness is attributed to the lower
loaf volume and denser texture. It was determined that as the PFD ratio
increased in breads, the volume values decreased (data not shown). The
denser texture formed by the decrease in volume may have caused an
increase in the hardness value (Supplementary material).
3.5. Sensory analysis
Sensory properties of bread samples are presented in Fig. 2. Ac­
cording to pseudocereal type, symmetry, pore structure and appearance
scores were higher in breads prepared with fermented dough of buck­
wheat and amaranth compared to breads prepared with fermented
dough of quinoa. Quinoa provided the highest taste-odor score in
breads, followed by buckwheat and amaranth. Although fermented
dough of amaranth added breads received high scores in terms of sym­
metry, pore structure and appearance, it was noteworthy that the taste-
odor scores were low. Similarly, Chlopicka et al. (2012) reported that
adding amaranth flour to wheat bread does not seem to be a favourable
modification due to its negative effect on taste. The difference in sym­
metry, pore structure, taste-odor, appearance and overall acceptability
scores according to pseudocereal type may be due to the natural char­
acteristic sensory properties of pseudocereal or may be due to different
factors affecting microbial growth in fermented dough. The availability
of nitrogen sources, carbohydrates, lipids as well as the buffer capacity,
enzymatic activity, and the presence of growth factors such as vitamins
and minerals as in the substrate can affect the microbial growth in
sourdoughs (Bender & Schönlechner, 2020). In terms of overall
acceptability, it was determined that 30% PFD ratio gave acceptable
results in gluten-free breads samples. When the sensory analysis results
were evaluated according to the fermentation method, it was seen that
the breads with PFD produced by yeast fermentation got significantly
higher scores in terms of symmetry, pore structure, taste-odor and
overall acceptability compared to spontaneous fermentation. Sourdough
is mainly used in wheat breads to enhance flavor, but the use of sour­
dough also has a significant impact on dough and final bread quality
(Arendt, Ryan, & Dal Bello, 2007). It has been reported by Karovičová
and Kohajdova (2007) that fermentation provides the development of a
wide variety of flavors, aromas and textures in foods.
4. Conclusion
The results of this study revealed that usage of PFD in gluten-free
bread formulation enhanced the ash, crude protein, TPC, AA and min­
eral content of bread samples. Breads containing fermented dough of
quinoa revealed higher TPC and AA than that of breads prepared with
fermented dough of buckwheat and amaranth. Addition of 45% PFD
markedly enhanced Ca (1.6 times), P (1.3 times), K (2.0 times), Fe (2.6
times), Mg (4.3 times) and Zn (2.4 times) contents of bread samples
compared to bread containing 0% PFD. Moreover, breads containing
fermented dough of quinoa showed the highest K content, and breads
prepared with fermented dough of amaranth showed the highest Ca and
Fe content. Fermentation of pseudocereals using bakers’ yeast resulted
7
LWT 160 (2022) 113301
in lower phytic acid content in breads compared to spontaneous
fermentation, but this difference was not significant (p>0.05). The uti­
lization of yeast in fermentation gave softer crumb texture in breads
compared to spontaneous fermentation. In sensory analyses, breads
containing buckwheat and amaranth fermented dough had higher
symmetry, pore structure, and appearance scores than breads containing
quinoa. Breads containing fermented dough of amaranth were less
appreciated in terms of taste-odor. Fermentation of pseudocereals using
bakers’ yeast was found to be more successful in improving sensory
properties of bread than spontaneous fermentation. Gluten-free bread
with acceptable quality can be produced by using up to 30% PFD. As a
result, by using pseudocereals fermented with bakers’ yeast in gluten-
free bread production, besides nutritional enrichment, both sensory
and textural improvement has been achieved.
Funding
This study was supported by the Karamanoğlu Mehmetbey Univer­
sity Scientific Research Projects Commission with the project no ’19-YL-
19’.
Declaration of competing interestCOI
The authors declare no conflict of interest relevant to this article.
CRediT authorship contribution statement
Saliha Yeşil: Investigation, Resources, Formal analysis, Writing –
original draft. Hacer Levent: Methodology, Project administration,
Conceptualization, Supervision, Writing – review & editing.
Declaration of competing interest
The authors declare no conflict of interest relevant to this article.
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