Journal of Cereal Science 102 (2021) 103315

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Journal of Cereal Science

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Dietary fibre, protein profile and technological characteristics of durum

spaghetti enriched with refined / whole grain hull-less barley flour
Dönüş Ermişer a, b, Erkan Yalçın b, *
a
Field Crops Central Research Institute, Ministry of Agriculture and Forestry, Ankara, Turkey
b
Department of Food Engineering, Bolu Abant İzzet Baysal University, Bolu, Turkey

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

Keywords: Durum wheat semolina (DWS) was substituted with refined or whole grain hull-less barley flour (RHBF or WHBF,
Spaghetti respectively) for producing nutritionally enriched spaghetti. The substitution levels were between 10 and 50%,
Hull-less barley and spaghetti made of DWS was used as control. Then, ash and protein contents, dietary fibre changes, protein
Dietary fibre
profile and technological characteristics were assessed. Total dietary fibre contents of DWS, RHBF and WHBF
SDS-PAGE
were 6.3, 7.6 and 20.8%, respectively. β-glucan contents of RHBF and WHBF were 2.40% and 5.0%, respectively.
Ash, protein, total-, soluble- and insoluble dietary fibres and β-glucan contents of uncooked spaghetti products
increased according to the control after hull-less barley flour (HBF) substitutions. Resistant starch (RS3) contents
of the control and HBF substituted spaghetti products considerably increased after cooking, the increment was
more noticeable in RHBF substituted ones. Brightness (L) and yellowness (b) values decreased and redness (a)
values increased while increasing of WHBF substitution. The total extractable proteins of uncooked spaghetti
products were investigated on sodium dodecyl sulphate polyacrylamide gel electrophoresis in order to monitor
the changes on polymeric protein distributions in HBF substituted spaghetti products. The HBF substitutions
could be an effective way to increase the dietary fibre intake, especially β-glucan and resistant starch.

1. Introduction
Pasta is a traditional cereal-based food product that is very popular
worldwide because of its convenience in terms of preparation in
different shapes and moods, nutritional quality, palatability and high
shelf-life duration. Durum wheat (Triticum durum) semolina is the raw
material of pasta products due to its unique bright yellow colour, high
protein, flavour and superior cooking quality, such as firmness, resistant
to surface disintegration and stickiness (Aalami et al., 2007; Sinesio
et al., 2008; Lamacchia et al., 2011).
The growing awareness of the relationship between diet and health
has led to the worldwide market trend for products with health benefits.
Whole grain cereal products are playing a key role in this trend and
constitute a growing focus for research to develop new value added
products (Ciccoritti et al., 2019). The epidemiological studies have
demonstrated that the consumption of whole grain foods could
contribute to an adequate intake of bioactive compounds, including
dietary fibres and antioxidant (Ciccoritti et al., 2019). Arndt (2006)
announced that incorporation of whole grain barley as part of a balanced
diet offers a variety of benefits, such as reducing the coronary heart
disease, type II diabets, certain cansers, and weight maintenance, due to
provide healthy and valuable phytonutrients.
Dietary fibres, which are polysaccharides of plant organs such as
cellulose, hemicellulose, lignin, β-glucan, are not digested and absorbed
in the human small intestine. They are threatened as complete or partial
fermentation in the large intestine. Dietary fibres have beneficial phys­
iological functions including laxation and improving bowel health by
stimulating growth of beneficial gut micro-flora, lowering blood
cholesterol and glucose levels, prevention of obesity, coronary heart
diseases, diabetes, blood pressure, lowering energy intake (Izydorczyk
et al., 2005; Sinesio et al., 2008; Aravind et al., 2012; Makhlouf et al.,
2019). These situations encourage people to consume more high fibre
cereal products, such as barley and oat. An interest in barley in human
nutrition is increasing due to be an excellent source of soluble and
insoluble dietary fibres, it is especially rich for β-glucan which is a sol­
uble dietary fibre (Sullivan et al., 2013). β-glucan is a nonstarch poly­
saccharide composed of linear chains of glucose with β-(1 → 3) and β-(1
→ 4) linkages found in the endosperm cell walls of cereals, mainly in
oats and barley (Tiwari and Cummins, 2009; De Paula et al., 2017).
Barley and oats, which are the primary sources of β-glucan in the human

* Corresponding author. Bolu Abant İzzet Baysal University Faculty of Engineering, Department of Food Engineering Gölköy Campus, 14030, Bolu, Turkey.
E-mail addresses: dermiser06@hotmail.com (D. Ermişer), yalcin_e@ibu.edu.tr (E. Yalçın).

https://doi.org/10.1016/j.jcs.2021.103315
Received 24 May 2021; Received in revised form 22 August 2021; Accepted 24 August 2021
Available online 27 August 2021
0733-5210/© 2021 Elsevier Ltd. All rights reserved.
D. Ermişer and E. Yalçın
diet, contain β-glucan in the ranges of 5–11% and 3–7%, respectively
(De Paula et al., 2017). The physiological functions of cereal β-glucan
are associated with viscosity, which in turn depends on the amount and
molecular weight of β-glucan solubilized in the intestine (De Paula et al.,
2017). The soluble fiber in whole-grains may help increase satiety by
delaying gastric emptying and slowing ingredient absorption in the
small intestine (Arndt, 2006). Izydorczyk et al. (2005) incorporated
fibre-enrich fractions of waxy and high amylose hull-less barleys into
Asian noodles for enrichment. They found that nutritional, cooking and
texture properties of the noodles could be improved or comparable to
noodles produced with wheat flours except for darkness of the colour.
Pasta is of nutritive importance for its non-cholesterol and low fat
content (Makhlouf et al., 2019). Pasta is also regarded as a product with
low glycaemic index, prompting a low postprandial blood glucose and
insulin response. Since, the compact structure of pasta can lead to a
dense protein network that causes reducing the starch hydrolysis
(Gelencser et al., 2008; Makhlouf et al., 2019). In addition, pasta is also
including resistant starch (RS) that occurs in cooking and then cooling
process (retrograde starch, RS3 type). RS, which is defined as starch
fractions resisting digestion in the small intestine, can be fermented by
gut micro-biota aiding short chain butyrate production that decreases
the faecal pH in the large intestine. RS acts like a dietary fibre which can
reduce the risk of the development of chronic diseases mentioned above
(Gelencser et al., 2008).
Pasta is a convenient food product for enriching with different food
sources in order to enhance nutritional quality and health benefits.
Therefore, the utilization of hull-less barley as an ingredient in pasta
products is preferred for its valuable sources of high protein, vitamins,
minerals, insoluble and soluble (β-glucan) dietary fibres and cherished
phytochemicals. Hull-less barley is harvested as naked and easy to mill
to fine either refine or whole grain flours (Yalçın et al., 2007). Barley
β-glucan is exerting important physiological benefits, such as preventing
coronary heart disease while reducing serum cholesterol level and
controllable blood glucose level (Sinesio et al., 2008; Aravind et al.,
2012; Montalbano et al., 2016; De Paula et al., 2017). Malcolmson et al.
(2014) stated that according to European Food Safety Assosciation
(EFSA), at least 3 g of barley β-glucans should be consumed per day to
obtain the health claimed effects. Previous studies showed that barley
flour or its β-glucan rich fractions or β-glucan concentrate could be
incorporated into pasta (Bourdon et al., 1999; Cleary and Brennan,
2006; Chillo et al., 2011; Aravind et al., 2012; Montalbano et al., 2016).
The main points of those studies were to investigate the carbohydrate
digestion which was slower in fibre-enriched pasta than that of pasta
made of durum semolina and also cholesterol lowering effect of barley
containing products. The authors also indicated that the incorporation
of fibre rich products into the protein-starch texture of wheat flour/se­
molina could alter the sensory and cooking properties of the final
product (Montalbano et al., 2016).
Therefore, the aim of the current research was mainly to study the
replacement of durum wheat semolina with increasing amount (from
10% to 50%) of refined or whole grain hull-less barley flour and to
determine the ash and protein contents, changes in the dietary fibre
composition, protein profiles and technological characteristics of the
spaghetti products. This research comes to the forefront with the study
of RS contents in uncooked and cooked spaghettis and also to examine
the protein profiles of uncooked spaghettis enriched with hull-less
barley flour on gel electrophoresis.
2. Material and methods
2.1. Materials
The native, two-rowed and spring type hull-less barley cultivar (cv.Ö
zen) and native durum wheat (cv. Eminbey) were provided from the Field
Crops Central Research Institute (Ankara, Turkey), were harvested from
Haymana (Ankara, Turkey) location in 2016. Durum wheat, which was
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Journal of Cereal Science 102 (2021) 103315
first conditioned to 16.5% moisture content, was roller-milled using
pilot scale mill (MLU 202, Buhler, Uzwil, Switzerland) in order to obtain
durum wheat semolina (DWS). Hull-less barley was first cleaned, then
conditioned to 15.5% moisture content and was milled to refined hull-
less barley flour (RHBF) using roller-mill (CD1 Mill, Chopin, Cedex,
France). The yield of RHBF was 30.1%. Wholegrain hull-less barley flour
(WHBF) was prepared using ultra-centrifuge mill (ZM 200, Retsch,
Germany) having 500 μ sieve.
2.2. Physical analysis
Thousand kernel weight (g) analyses was carried out first deter­
mining the number of kernels in 20 g clean sample, and then converted
to 1000 kernels, the results expressed on dry weight basis. Kernel
hardness (%) and test weight (kg/hL) was determined according to
Approved Methods of Analysis Method No. 55–31.01 and 55–10.01,
respectively, (AACCI, 2010); and vitreousness (%) was determined ac­
cording to ICC Standard Method No.129 (ICC, 2002). Particle size
measurements of DWS, RHBF and WHBF samples were carried out in the
equipment of Mastersizer, 2000 Version 5.60 (Malvern Instruments Ltd.,
UK).
2.3. Physicochemical analysis of DWS
Wet gluten and gluten index values were determined according to
Approved Methods of Analysis Method No: 38–12.02 (AACCI, 2010).
After drying the wet gluten in glutork (Perten, Switzerland) equipment,
dry gluten content was determined. Wet gluten was also tested in
glutograph-E (Brabender, Duisburg, Germany) equipment in order to
determine the “stretching capability” and “extensibility time” values.
Determination of sedimentation value in sodium dodecyl sulphate (SDS)
solution and modified (delayed) SDS sedimentation value was done
according to Williams et al. (1986).
2.4. Chemical analysis
Moisture, crude protein (%N*5.7 for DWS, %N*6.25 for hull-less
barley flours) and ash contents in raw materials and spaghetti were
determined according to Approved Methods of Analysis Methods No.
44–15.02, 46–30.01, and 08–01.01 (AACCI, 2010), respectively. The
determination of β-carotene contents in DWS and hull-less barley flours
were accomplished following the Approved Methods of Analysis Method
No. 14–50.01 (AACCI, 2010).
2.5. Blending of DWS and hull-less barley flour
Durum wheat semolina (DWS) was substituted with RHBF or WHBF
at the levels of 10, 20, 30, 40 and 50% (w/w). The blending of DWS and
RHBF or WHBF was first carried out in a covered container. Then each
blend was homogenized in dough kneader (Namad, Italy) without water
for 5 min. After homogenization, the blends were kept in a covered
container until mixing with water.
2.6. Farinograph analysis
The farinograph properties of raw materials, like DWS and hull-less
barley flour (HBF) blends were determined according to Approved
Methods of Analysis Method No: 54–21.01 (AACCI, 2010) using Bra­
bender Farinograph-E (Brabender OHG, Duisburg, Germany). For the
production of control spaghetti from DWS, the water content in dough
was considered as 31%. An amount of water (mL), which was added into
DWS or blends, was calculated according to their water absorption
values detected in farinograph analysis, the calculation shown below:
(31 × F(%)) 1000
Amount of added water (mL) = × (1)
A (%) 100
D. Ermişer and E. Yalçın
F: Water absorption values of hull-less barley flour blends (%)
A: Water absorption value of DWS (53.7%)
1000: 1000 g of each control or hull-less barley flour incorporated
blend.
2.6.1. Spaghetti production
The spaghetti was produced according to the method of D’Egidio
et al. (1982) by using pilot-scale pasta making machine (Namad, Rome,
Italy). First, the DWS and RHBF or WHBF incorporated blends (10, 20,
30, 40, 50%) were mixed with water in dough kneader in order to obtain
homogenized pasta dough. For this, the calculated water amount was
added into DWS or HBF blends at 40 ◦ C. The control dough and blends
were mixed in a vacuum (700 mm-Hg) for 15 min. Then, the spaghetti
dough was pressed into the extruder at 600 mm-Hg vacuum pressure
and at 45 ◦ C temperature. The control and HBF incorporated spaghetti
samples were collected from the extruder die (1.7 mm diameter) and
then the low temperature (40 ◦ C) drying process was applied at 60%
relative humidity. After lowering the moisture content of spaghetti
samples below 12%, they were cooled to room temperature and were
kept in plastic bags. Some of spaghetti products were milled into the fine
flour by using ultra-centrifuge mill for analysis. All spaghetti samples
were prepared at three batches and analysed separately.
2.7. Determination of total, soluble and insoluble dietary fibres
Total dietary fibre (TDF), soluble and insoluble dietary fibre (SDF,
IDF, respectively) contents of DWS, RHBF, WHBF and uncooked spa­
ghetti products were determined by the enzymatic-gravimetric method
using Megazyme Total Dietary Fibre Assay Kit (Megazyme International
Ireland Limited, Wicklow, Ireland). The average values (%) of trplicate
analysis were reported on dry weight basis.
2.8. Determination of β-glucan
β-glucan contents of raw materials and uncooked spaghetti products
were determined by using Megazyme Mixed-Linkage β-glucan Assay Kit.
β-glucan analysis was performed as triplicate analysis and the average
value (%) was reported on a dry weight basis.
2.9. Determination of resistant starch
Resistant starch contents of uncooked and cooked spaghetti products
were determined by using Megazyme Resistant Starch Assay Kit. All
cooked spaghetti samples were prepared at their optimum cooking time,
and then they were frozen to − 18 ◦ C in order to freeze-dry and then they
were kept at +4 ◦ C until analysis. Resistant starch analysis was per­
formed as triplicate analysis and the average value (%) was reported on
dry weight basis.
2.10. Colour measurements
Colour properties of uncooked dry spaghetti samples were measured
using a bench-top colour spectrophotometer (Gardner Color View, BYK-
Instruments, Germany). Uncooked spaghetti sticks were filled into the
place of equipment without any empty space. Colour properties were
expressed according to Hunter-Lab system, like “L” value [0–100:
darkness to brightness]; “a” value [-a: greenness, + a: redness]; “b”
value [-b: blueness, + b: yellowness]. The average value of the triplicate
analysis was reported for each sample with standard deviation.
2.11. Cooking properties
Optimum cooking time of spaghetti (the time that is necessary to
obtain complete gelatinization of starch) was determined due to
Approved Methods of Analysis Method No. 16–50.01 (AACCI, 2010).
For cooking process, 25 g of spaghetti was put into 300 mL of boiling tap
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Journal of Cereal Science 102 (2021) 103315
water with no salt addition. Then, the optimal cooking time of spaghetti
was determined until disappearance of the continuous white line visible
at the centre of a crushed piece of spaghetti using a Plexiglas crushing
plate. The cooking loss (%), water absorption (%), volume increase (%)
and total organic matter (g starch/100 g) were determined according to
the method of D’Egidio et al. (1982).
2.12. Textural properties
The analysis of the textural characteristics of the spaghetti was
performed using a Stable Micro System (SMS) (TA-XT2i, Surrey, UK).
The optimum cooked spaghetti was drained and subsequently used for
instrumental texture analysis. Five replicates of each test were done in
order to calculate the average value. The following parameters were
determined: Breaking force and Breaking distance: These parameters were
determined using a flexure rig (code: A/SFR) of SMS equipment. The
uncooked (dry) spaghetti samples had a length of 20 cm and then they
were placed between load cell (5 kg) and lower plate of the device. The
pre-test speed, test speed and post-test speed were adjusted to 0.5 mm/s,
2.5 mm/s and 10 mm/s, respectively. The breaking force (g) and
breaking distance (mm) of the spaghetti were measured while the upper
head of the device was moving towards down. Firmness: The test was
done according to Approved Methods of Analysis Method No. 66–52.01
(AACCI, 2010) and Sinesio et al. (2008). Two strands of cooked spa­
ghetti (the length of each was 5 cm) were aligned side by side on the
plate of the SMS instrument. The instrument was equipped with a
Perspex blade (1 mm thickness; code A/LKB-F) to cut the spaghetti
strands. The blade was lowered into the sample until 0.5 mm distance
left to lower plate at a crosshead speed of 0.17 mm/s. The speed of
post-experiment was 10 mm/s. The area under the curve was used as a
measure of firmness (g.cm). Adhesiveness: Five strands of cooked spa­
ghetti (the length of each was 10 cm) were aligned side by side on the
plate of the SMS instrument. The instrument was equipped with a
plunger (38 × 50 mm2 contact surface; code HDP/PFS). A Plexiglas plate
with an opening (40 × 52 mm2) for plunger-to-sample contact was
placed on the spaghetti as a sample retainer. The weight of the plate was
sufficient to prevent changes in the spaghetti alignment. The plunger
moved down at a speed of 0.5 mm/s. Once a compression force of 1000 g
was reached, the plunger stopped for 2 s and then lifted from the surface
at a speed of 10 mm/s. The maximum tensile force (g.s) recorded during
the separation of the plunger was taken as a measure of adhesiveness
(Sinesio et al., 2008). Elasticity: One end of a strand of cooked spaghetti
(the length of each was 25 cm) was fixed at the crosshead and the other
fixed to the instrument platform using tensile grips (code A/SPR). The
test was carried out at a crosshead speed of 3 mm/s. The elastic modulus,
the force at the strand rupture (tensile strength, g), and the elongation at
the strand rupture (g) were measured (Sinesio et al., 2008).
2.13. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-
PAGE)
SDS-PAGE was performed based on the original procedure of
Laemmli (1970) modified by Fu and Sapirstein (1996). In order to get
equal concentrations of proteins, DWS, RHBF, WHBF and uncooked
spaghetti were weighed on the basis of their dry weight protein contents
and mixed with 1 mL of buffer solution (pH 6.8) containing 0.063 mol/L
Tris-HCl, 2% (w/v) SDS, 20% (w/v) glycerol (Merck, Germany) and
0.01% (w/v) pyronine Y (Sigma-Aldrich, USA). The reduced condition
was created by adding 5% (v/v) β-mercaptoethanol (2-β-ME,
Sigma-Aldrich, USA) into buffer solution. The blend was vortexed (Reax
Top model, Heidolph, Germany) for 1 min in every 10 min during 2 h.
Extracted and dissolved proteins were heated on dry block heating
thermostat (Bio TDB-120 model, BIOSAN, Latvia) for 3 min in order to
denature proteins before applying (10 μL) to the stacking gel. SDS-PAGE
was carried out in a cooled (15 ◦ C) slab gel unit (Protean II xi Cell,
Bio-Rad, CA, USA). The acrylamide concentrations of stacking gel and
D. Ermişer and E. Yalçın Journal of Cereal Science 102 (2021) 103315

resolving gel were 5% and 12.5%, respectively. After concluding the
electrophoresis, the gels were rinsed in rinsing solution (57% (v/v)
water+ 33% (v/v) methanol+ 10% (v/v) trichloroacetic acid (100%
w/v)) overnight. Then, the gels were stained overnight with Coomassie
brillant blue G-250 (Merck, Darmstadt, Germany) according to Ng and
Bushuk (1987). Apparent molecular weights were determined using the
wide range molecular weight protein markers (Sigma S8445, MO, USA).
The determination of the molecular weights of the protein bands were
carried out by using Bio-Rad Image Lab 5.0 software after scanning from
the gel imager (ChemiDoc™ MP Imaging System, Bio-Rad, CA, USA).
2.14. Statistics
All data, which were presented as means with standard deviations,
were analysed using JMP 13 statistical software (SAS Institute Inc., USA)
and were evaluated according to factorial order (2x5 factorial) in ran­
domized blocks experimental design. The analysis of variance (ANOVA)
was applied and the difference among means were compared using the
Least Significance Difference (LSD) test with the confidential levels of
99% (significance level of p < 0.01) or 95% (significance level of p <
0.05). The coefficients of variances (CV) were also reported.
3. Results and discussion
The test weight, thousand kernel weight, kernel hardness and vit­
reousness of hull-less barley were 76.8 kg/hL, 31.9 g, 40.0% and 2.0%,
respectively. They were 78.4 kg/hL, 36.2 g, 70.0%, and 99.0% for the
durum wheat, respectively. The average particle size (d(0.90)) results of
DWS, RHBF and WHBF were 472 μ, 168 μ, 476 μ, respectively. The wet
gluten, dry gluten, gluten index, stretching capability and extensibility
time of DWS were found as an average of 31.7%, 10.2%, 98.1%, 244 BU
and 125 s, respectively. The SDS and modified SDS sedimentation values
of DWS were 17 mL and 18 mL, respectively, on 14% moisture basis.
Water absorption (%) value determined in farinograph and calcu­
lated added water amount (mL) of DWS were found as 53.7% and 310
mL, respectively. Water absorption values determined in farinograph
and calculated added water amounts (mL) of RHBF incorporated
(10–50%) blends were in the ranges of 53.3–54.7% and 308–316 mL,
respectively. Besides, these values in WHBF incorporated (10–50%)
blends were in the ranges of 55–64.8% and 318–374 mL, respectively.
The final percentages of water content in the RHBF or WHBF incorpo­
rated spaghetti dough were in the ranges of 30.8–31.6% and
31.8–37.4%, respectively.
Some chemical properties of DWS, RHBF and WHBF were shown in
Table 1. The highest protein content was found in the WHBF (18.2%).
The protein content of DWS (12.4%) was slightly higher than that of the
RHBF (12.1%). Durum wheat semolina had the highest β-carotene
content (7.7 mg/kg) among the samples. The WHBF had the highest
TDF, IDF and SDF contents. It can be indicated that the IDF content of
Table 1
Some chemical properties of durum wheat semolina (DWS), refined hull-less
barley flour (RHBF) and whole grain hull-less barley flour (WHBF).
Chemical properties DWS RHBF WHBF
Moisture (%) 12.3 ± 0.10 13.2 ± 0.05 9.5 ± 0.07
Ash (%) 0.63 ± 0,006 0.93 ± 0,018 2.00 ± 0,000
Protein (%) 12.4 ± 0,48 12.1 ± 0,44 18.2 ± 0,35
β-carotene (mg/kg) 7.7 ± 0.05 2.6 ± 0.06 5.0 ± 0.39
TDF (%) 6.3 ± 0.09 7.6 ± 0.30 20.8 ± 0.23
IDF (%) 3.1 ± 0.15 2.5 ± 0.23 11.8 ± 0.37
SDF (%) 3.2 ± 0.18 5.1 ± 0.07 9.0 ± 0.53
β-Glucan (%) 0.01 ± 0.011 2.40 ± 0.051 5.00 ± 0.040
Resistant Starch (%) 0.67 ± 0.038 0.26 ± 0.045 0.40 ± 0.030
Each parameter was analysed triplicate (n = 3). The results were given as mean
± standard deviation on dry weight basis. TDF: Total dietary fibre, IDF: Insoluble
dietary fibre, SDF: Soluble dietary fibre.
DWS (3.1%) was higher than that of the RHBF (2.5%), but lower than
that of the WHBF (11.8%). The β-glucan contents of the RHBF and
WHBF flours were found as 2.4% and 5.0%, respectively. Besides, the
highest resistant starch (RS) content was found in the sample of DWS as
0.67%.
Some chemical properties of uncooked spaghetti products were
shown in Table 2. The final moisture contents of the spaghetti products,
which did not change considerably, were between 9.3% and 9.9%. The
ash contents of the RHBF substituted (from 10 to 50%) uncooked spa­
ghettis were between 0.67 and 0.73% (p < 0.01). Besides, the ash con­
tents of the WHBF substituted ones were higher than that of the control
and RHBF substituted spaghetti samples (p < 0.01). The ash contents
increased with the increasing of the WHBF substitution from 10%
(0.80%) to 50% (1.30%, p < 0.01). The ash contents of the RHBF or
WHBF substituted spaghetti products were within Italian legal limits for
whole grain pasta (1.80 g/100 g) as indicated by Ciccoritti et al. (2019).
The protein contents of WHBF substituted spaghetti products were
higher than that of the control and the RHBF substituted spaghetti
samples (p < 0.01), the protein contents changed between 12.3% and
13.0% in the WHBF substituted samples where the proteins slightly
increased with an increasing of substitution level. The protein contents
did not change significantly in both the control and RHBF incorporated
ones (p > 0.01).
The colour properties of uncooked spaghetti products were also
shown in Table 2. The brightness (L) values of the control and RHBF
replaced spaghetti samples were mostly similar to each other (p > 0.01)
and they were higher than that of the WHBF substituted ones whose
brightness values did not significantly change while increasing the
substitution level (p > 0.01) except for 50% WHBF substituted spaghetti
which had the lowest brightness value (p < 0.01). Makhlouf et al. (2019)
also found that an increasing incorporation level of whole grain barley
flour into fusilli pasta caused a decrease in brightness value. The redness
(a) values of the WHBF substituted spaghetti samples, which increased
while increasing the substitution level, were higher (p < 0.01) than that
of the control and RHBF replaced spaghetti products whose redness
values were similar to each other (p > 0.01). The yellowness (b) value of
the control spaghetti was 24.2 which was the highest value among all
samples (p < 0.01). The yellowness values decreased while increasing
the both types of hull-less barley flour substitution levels and that
reduction was more noticeable in WHBF substituted uncooked spaghetti
products (p < 0.01). Izydorczyk et al. (2005) specified that discoloration
of food products containing barley occurs due to the complex oxidative
polymerization of the several phenolic compounds present in this grain.
The dietary fibre contents of uncooked spaghetti products were
shown in Table 3. IDF, SDF and TDF contents of RHBF substituted spa­
ghetti products did not change significantly (p > 0.01) while increasing
the substitution level from 10% to 50%, whereas IDF, SDF and TDF
contents of WHBF substituted spaghetti products significantly increased
with the increasing substitution levels (p < 0.01). The IDF, SDF and TDF
contents in all barley flour substituted spaghetti products were in the
ranges of 3.1–7.2%, 3.3–6.2% and 6.3–13.5%, respectively. The control
spaghetti and 50% WHBF substituted spaghetti had the lowest and
highest dietary fibre contents, respectively (p < 0.01). The β-glucan
contents of both RHBF and WHBF substituted spaghetti samples
increased while increasing the substitution level from 10% to 50%, be­
sides their values changed in the ranges of 0.40–1.02% and 0.47–2.38%,
respectively. The WHBF substituted spaghetti samples had higher con­
tents of β-glucan than that of the RHBF substituted ones. The highest and
the lowest β-glucan contents were seen in the spaghetti samples of 50%
WHBF substituted spaghetti (2.38%) and the control sample (0.20%),
respectively (p < 0.01). De Paula et al. (2017) found the β-glucan con­
tents in dried uncooked pastas produced with waxy hull-less barley flour
at the levels of 30%, 50% and 100% as 2.93%, 5.0% and 9.72%,
respectively.
The resistant starch (RS) contents of uncooked and cooked spaghetti
products prepared by replacing DWS with different levels (10–50%) of

4
D. Ermişer and E. Yalçın Journal of Cereal Science 102 (2021) 103315

Table 2
Some chemical and colour properties of uncooked (dry) spaghetti products.
SL (%) Moisture (%) Ash* (%) Protein* (%) L, brightness a, redness b, yellowness

Control 0 9.3 ± 0.35 0.60 ± 0.000h 11.8 ± 0.26bc 51.7 ± 0.63 ab 2.60 ± 0.083e 24.2 ± 0.26a
RHBF 10 9.3 ± 0.12 0.67 ± 0.058g 11.9 ± 0.29bc 52.5 ± 1.14 ab 2.58 ± 0.277e 22.8 ± 0.25b
20 9.4 ± 0.00 0.70 ± 0.000 fg 11.7 ± 0.44bc 50.0 ± 1.73bc 2.27 ± 0.125e 21.7 ± 0.16c
30 9.5 ± 0.58 0.70 ± 0.000 fg 11.3 ± 0.06cd 51.9 ± 1.16 ab 2.44 ± 0.180e 20.8 ± 0.25d
40 9.6 ± 0.06 0.73 ± 0.058f 11.2 ± 0.35cd 54.2 ± 0.91a 2.43 ± 0.244e 20.4 ± 0.09d
50 9.9 ± 0.06 0.70 ± 0.000 fg 10.7 ± 0.10d 53.5 ± 0.34a 2.63 ± 0.309e 19.3 ± 0.07e
WHBF 10 9.9 ± 0.00 0.80 ± 0.000e 12.3 ± 0.25 ab 48.3 ± 1.05cd 3.72 ± 0.439d 21.1 ± 0.07cd
20 9.9 ± 0.10 0.90 ± 0.000d 12.8 ± 0.15a 47.9 ± 1.81cd 4.74 ± 0.101c 19.3 ± 1.04e
30 9.8 ± 0.00 1.00 ± 0.000c 12.7 ± 0.68a 46.1 ± 0.98d 5.37 ± 0.248b 18.4 ± 0.41f
40 9.6 ± 0.06 1.13 ± 0.058b 13.0 ± 0.40a 46.4 ± 0.68d 5.96 ± 0.373b 18.6 ± 0.21ef
50 9.8 ± 0.06 1.30 ± 0.000a 13.0 ± 0.31a 42.5 ± 0.74e 6.78 ± 0.406a 16.5 ± 0.17g
LSD 0.066 0.77 2.585 0.612 0.841
CV 0.02 0.035 0.33 0.023 0.073 0.018

Each parameter was analysed triplicate (n = 3). The results were given as mean ± standard deviation. *Reported on dry weight basis.
Values followed by the different letters in the same column are significantly different (p < 0.01).
SL: Substitution Level; RHBF: Refined Hull-less Barley Flour; WHBF: Whole grain Hull-less Barley Flour.
LSD: Least Significance Difference, CV: Coefficient of Variance.

Table 3
Dietary fibre compositions of uncooked (dry) and resistant starch contents of uncooked and cooked spaghetti products.
SL (%) IDF* (%) SDF* (%) TDF* (%) β-glucan* (%) RS**,a (%) RS3**,b (%)

Control 0 2.5 ± 0.25g 3.2 ± 0.17d 5.8 ± 0.06ı 0.20 ± 0.015h 0.44 ± 0.040c 2.61 ± 0.04c
RHBF 10 3.1 ± 0.12f 3.3 ± 0.32cd 6.4 ± 0.26gh 0.40 ± 0.020g 0.45 ± 0.020c 2.80 ± 0.042b
20 3.0 ± 0.17 fg 3.3 ± 0.17cd 6.3 ± 0.25h 0.61 ± 0.030f 0.60 ± 0.032b 2.79 ± 0.037b
30 3.0 ± 0.06 fg 3.7 ± 0.15cd 6.6 ± 0.17fgh 0.70 ± 0.045ef 1.39 ± 0.047a 3.40 ± 0.037a
40 3.2 ± 0.15ef 3.8 ± 0.10c 7.0 ± 0.06f 0.81 ± 0.110e 0.64 ± 0.053b 2.51 ± 0.042d
50 3.0 ± 0.12 fg 3.8 ± 0.10c 6.8 ± 0.06 fg 1.02 ± 0.010d 0.32 ± 0.030d 2.45 ± 0.024d
WHBF 10 3.6 ± 0.32e 3.8 ± 0.06c 7.4 ± 0.23e 0.47 ± 0.046g 0.66 ± 0.021b 2.75 ± 0.052b
20 4.3 ± 0.44d 4.9 ± 0.23b 9.2 ± 0.15d 0.95 ± 0.040d 0.63 ± 0.036b 2.45 ± 0.024d
30 5.4 ± 0.26c 5.3 ± 0.42b 10.6 ± 0.12c 1.38 ± 0.060c 0.46 ± 0.031c 1.95 ± 0.034e
40 6.2 ± 0.00b 5.2 ± 0.17b 11.4 ± 0.17b 1.83 ± 0.067b 0.37 ± 0.036d 1.80 ± 0.026f
50 7.2 ± 0.12a 6.2 ± 0.32a 13.5 ± 0.21a 2.38 ± 0.119a 0.36 ± 0.031d 1.36 ± 0.056g
LSD 0.50 0.51 0.38 0.135 0.042 0.067
CV 0.10 0.05 0.02 0.064 0.063 0.016

Each parameter was analysed triplicate (n = 3). The results were given as mean ± standard deviation on dry weight basis. Values followed by the different letters in the
same column are significantly different *(p < 0.01); **(p < 0.05). TDF: Total dietary fibre; IDF: Insoluble dietary fibre; SDF: Soluble dietary fibre.
SL: Substitution Level; RHBF: Refined Hull-less Barley Flour; WHBF: Whole grain Hull-less Barley Flour.
LSD: Least Significance Difference; CV: Coefficient of Variance.
a
Uncooked spaghetti.
b
Cooked spaghetti.

RHBF or WHBF were also shown in Table 3. The RS contents of all un­
cooked spaghetti products were below 1% except for 30% RHBF
substituted spaghetti which had the highest (1.39%, p < 0.05) among
the uncooked samples. There was first an increment in the RS contents of
RHBF substituted spaghetti products and then RS contents decreased to
0.32% (p < 0.05). Besides, the RS contents of the uncooked spaghetti
samples including WHBF decreased consistently from the substitution
levels of 10% (0.66%) to 50% (0.36%). Gelencsér et al. (2008) reported
that RS3 is a retrograded starch (mainly amylose) and can occur in
cooked and cooled spaghetti samples. After cooking, the RS3 contents of
cooked spaghetti samples increased considerably at each substitution
level, this increment was also observed in the control spaghetti (2.61%).
The highest RS3 content was also seen in the cooked sample of 30%
RHBF substituted spaghetti as 3.40% (p < 0.05). The RS3 contents of
cooked RHBF substituted spaghetti samples were changed in the range
of 2.45–3.40%. It was an interesting that the RS3 contents of cooked
WHBF substituted spaghetti samples decreased from 2.75% to 1.36%
while increasing the substitution level (p < 0.05). The increments of RS3
contents in the RHBF (10–30%) substituted spaghetti products after
cooking was higher than that of the corresponding WHBF substituted
ones.
Tiwari and Cummins (2009) stated that the Food and Drug Admin­
istration (FDA) of US reported health claims for β-glucan-rich cereal
grains, for oat grains, at least 0.75 g β-glucan/per serving can make the
health claim that they may reduce the risk of heart disease and FDA also
recommended minumum 10% TDF content of milled hull-less barley
allowable health claim. In this study, the spaghetti produced with 10%
RHBF substitution provided 2.8 g SDF, 0.34 g β-glucan, 5.4 g TDF and
0.38 g RS/per serving size of 85 g dry spaghetti and 2.38 g RS3/per
serving size of 85 gr cooked spaghetti. The SDF content meets the
criteria of the FDA and all amounts of per serving size were higher than
that of the control spaghetti. Besides, the spaghetti produced with 10%
WHBF substitution provided 3.2 g SDF, 0.40 g β-glucan, 6.3 g TDF and
0.56 g RS/per serving size of 85 g dry spaghetti and 2.34 g RS3/per
serving size of 85 gr cooked spaghetti. Malcolmson et al. (2014) also
indicated that the ability to make a barley health claim would be
influenced by the product type and level of barley flour used, the variety
of barley and the country where the claim would be made.
The cooking properties of spaghetti products were shown in Table 4.
The highest volume increase (240%) was obtained with the 40% RHBF
substituted spaghetti followed by the control (233%, p > 0.01). The
volume increases of the RHBF substituted spaghetti samples raised while
increasing the substitution level, however it decreased at the 50% sub­
stitution level. An increasing of RHBF incorporation up to 50% may
cause an increase in fibre solubilisation that may form viscous network
restricting volume increase (Izydorczyk et al., 2005). Aravind et al.
(2012) also explained that the preferential hydration of β-glucan over
starch and gluten is limiting both the development of the gluten network

5
D. Ermişer and E. Yalçın Journal of Cereal Science 102 (2021) 103315

Table 4
Cooking properties of hull-less barley flour substituted spaghetti products.
SL (%) Volume Increase** (%) Water Absorption* (%) Optimum Cooking Time** (min) Cooking Loss (%) TOM** (g starch/100g)

Control 0 233.3 ± 14.43 ab 185.7 ± 8.08 ab 12.4 ± 0.17de 6.5 ± 0.47 1.73 ± 0.058d
RHBF 10 208.7 ± 7.51bcd 178.7 ± 6.66abc 13.1 ± 0.06cde 6.9 ± 0.40 2.00 ± 0.100cd
20 215.0 ± 8.66bcd 182.3 ± 4.04 ab 14.2 ± 0.71 ab 6.4 ± 0.15 2.43 ± 0.289bc
30 225.0 ± 0.00abc 186.3 ± 4.73 ab 15.0 ± 0.444a 7.1 ± 0.30 2.36 ± 0.404bc
40 240.0 ± 0.00a 192.0 ± 12.12a 13.2 ± 0.58cd 7.8 ± 0.06 2.66 ± 0.153 ab
50 225.0 ± 0.00abc 188.0 ± 4.36 ab 12.1 ± 0.47ef 7.9 ± 0.36 3.06 ± 0.058a
WHBF 10 208.3 ± 14.43cd 176.0 ± 4.36bcd 14.3 ± 0.00 ab 6.7 ± 0.21 2.23 ± 0.503bcd
20 173.3 ± 12.50e 165.0 ± 14.80de 14.9 ± 0.40a 7.2 ± 0.57 2.00 ± 0.100cd
30 147.3 ± 20.98f 168.3 ± 6.51cde 13.5 ± 0.10bc 7.4 ± 0.25 1.90 ± 0.000cd
40 114.3 ± 7.51g 167.3 ± 4.04cde 12.3 ± 0.06def 7.1 ± 0.55 2.10 ± 0.265bcd
50 200.0 ± 0.00d 158.0 ± 10.58e 11.3 ± 0.85f 7.9 ± 0.29 1.66 ± 0.416d
LSD 24.75 13.66 0.98 0.588
CV 0.05 0.05 0.03 0.05 0.119

Each parameter was analysed triplicate (n = 3). The results were given as mean ± standard deviation. Values followed by the different letters in the same column are
significantly different (*p < 0.05; **p < 0.01).
SL: Substitution Level; RHBF: Refined Hull-less Barley Flour; WHBF: Whole grain Hull-less Barley Flour; TOM: Total Organic Matter.
LSD: Least Significance Difference; CV: Coefficient of Variance.

via minimizing water absorption and finally starch swelling. Besides, the
counter effect was seen with the WHBF substituted spaghetti samples.
The control and RHBF substituted spaghetti samples had similar (p >
0.05) water absorption (%) values, however that parameter was lower in
the samples of WHBF substituted spaghetti samples than that of those (p
< 0.05). The optimum cooking time of the control spaghetti was 12.4
min. It changed in the range of 11.3–15.0 min in the hull-less barley
flour substituted spaghetti products. Lamacchia et al. (2011) explained
that the optimal cooking time increased as protein and β-glucan contents
increased, due to their reactivity with water. Furthermore, the addition
of higher amount of WHBF disrupted the continuity of the protein matrix
of DWS, which probably resulted in faster heat/moisture penetration
and shorter cooking time (Izydorczyk et al., 2005). The cooking loss of
the control (6.5%) was generally lower than the hull-less barley flour
substituted spaghetti products. Montalbano et al. (2016) declared that if
the cooking loss value is lower than 7%, pasta is good quality. The
cooking loss slightly increased with the increasing of both types of
hull-less barley flour substitution. The cooking loss (7.9%) of the 50%
hull-less barley flour substituted products was found as the highest.
However, the cooking loss values of all spaghetti products were much
lower than that of the values determined in the study of Makhlouf et al.
(2019) where whole grain barley flour incorporation (5, 10, 15%) into
fusilli pasta performed. The high cooking loss of the durum wheat pasta
could be explained by the disruption of the protein network through the
cooking process and releasing exudates during starch gelatinization,
which causes an increase in cohesiveness and stickiness on the surface of
the cooked pasta (Gelencsér et al., 2008; Montalbano et al., 2016). The
total organic matter (TOM) value of the control (1.73 g starch/100 g)
was lower than that of the hull-less barley flour substituted spaghetti
samples except for the 50% WHBF substituted spaghetti whose TOM
value (1.66 g starch/100 g, p > 0.01) was found as the lowest among all
samples. Generally, while the TOM values of the RHBF substituted
spaghetti products slightly increased with the increasing of substitution
level. The 50% RHBF substituted spaghetti had the highest TOM value as
3.06 g starch/100 g among all samples. Contrary, the TOM values of the
WHBF substituted samples generally followed decreasing trend in this
study. Lamacchia et al. (2011) clarified that partial or complete substi­
tution of durum wheat semolina with fibre material can result in nega­
tive changes for pasta quality, like increased cooking loss and softening.
They found that increasing the level of β-glucan (up to 10%) in the pasta
was able to prevent excessive cooking loss due to polysaccharide
network, capable of encapsulating the protein-starch matrix and
strengthening the structural integrity of the pasta. In addition, the high
water-binding capacity of β-glucan caused a restriction of starch
swelling and gelatinization, an increase of the gelatinization tempera­
ture that resulted in an increase in optimum cooking time. Malcolmson
et al. (2014) explained for their study that barley flours typically have a
much greater water absorption capacity than wheat flours due to their
higher levels of soluble fibre, mainly β-glucans, which could affect
baking performance.
The textural properties of spaghetti samples were shown in Table 5.
Breaking force, firmness and elasticity values of the control spaghetti
were higher than that of hull-less barley flour substituted ones. Breaking
force, breaking distance, firmness and elasticity values of RHBF
substituted spaghetti products decreased, conversely the adhesiveness
slightly increased while increasing the substitution level. Reduced
firmness of the spaghetti matrix caused possibly due to disruption of the
gluten matrix by inclusion of fibre rich hull-less barley flour (Izydorczyk
et al., 2005). The textural properties of WHBF substituted spaghetti
samples did not generally follow regular trend except for breaking dis­
tance and firmness values which slightly increased and decreased,
respectively, while increasing of the substitution level. The breaking
distance and adhesiveness values of the WHBF substituted spaghetti
products were obviously lower than that of the RHBF substituted ones (p
< 0.01). The other textural properties of the spaghetti samples produced
with hull-less barley flours were closed to each other. Makhlouf et al.
(2019) also found that an increasing levels of whole grain barley flour in
fusilli pasta caused significant decrease in pasta firmness and adhe­
siveness. Lamacchia et al. (2011) indicated that pasta firmness was
positively affected by total protein content and negatively affected by
β-glucans and large polymeric proteins. The large aggregates of poly­
meric proteins and the new bonds formed between durum wheat
semolina and barley proteins may form a stronger gluten network due to
S–S bonds, allowing starch to absorb less water, preventing pasta
stickiness and increasing firmness.
Total extractable proteins of DWS, RHBF, WHBF and uncooked
spaghetti products in non-reducing and reducing conditions were shown
in Fig. 1A and B, respectively. In this electrophoregrams, groups of
wheat and barley proteins were termed as described in Tatham et al.
(2000). Graziano et al. (2019) and Visioli et al. (2018) stated that the
technological and nutritional properties of durum wheat semolina
depend mainly on the protein content and gluten characteristics, which
are affected by genotype, environment and processing conditions.
Durum wheat gluten, which consists of gliadin (prolamin type) and
glutenin (glutelin type) proteins, influences dough strength, extensi­
bility (gliadin), elasticity (glutenin) and stability. The amino acid
composition of gluten proteins also makes them highly resistant to
gastric and pancreatic proteases (Graziano et al., 2019). Therefore, in
this study, the changes on protein profiles of hull-less barley substituted
spaghetti products were examined.
The electrophoregram of the non-reduced samples showed that the
high molecular weight (HMW, more than 200 kDa) proteins could not

6
D. Ermişer and E. Yalçın Journal of Cereal Science 102 (2021) 103315

Table 5
Textural properties of uncooked (dry) and cooked spaghetti products.
SL (%) Breaking Force** (g) Breaking Distance** (mm) Firmness* (g.cm) Adhesiveness** (g.s) Elasticity** (g)

Control 0 52.7 ± 0.69a 27.9 ± 0.82b 8.4 ± 0.35a 121.6 ± 3.16c 31.3 ± 1.37a
RHBF 10 50.3 ± 0.91bc 35.4 ± 4.88a 7.8 ± 0.31bc 133.3 ± 1.24b 27.1 ± 1.02bc
20 47.3 ± 0.07ef 23.8 ± 0.55bc 7.7 ± 0.04bc 134.9 ± 0.30b 24.7 ± 1.04cd
30 48.1 ± 0.16de 30.1 ± 4.53 ab 6.8 ± 0.48d 134.5 ± 2.46b 21.8 ± 0.66de
40 45.6 ± 1.02 fg 16.5 ± 4.42cd 6.2 ± 0.33e 138.4 ± 0.49b 19.8 ± 3.111e
50 45.3 ± 1.94g 15.5 ± 2.74de 6.0 ± 0.17e 147.4 ± 1.34a nd
WHBF 10 49.1 ± 0.31cd 9.1 ± 2.28e 8.1 ± 0.36 ab 93.7 ± 8.31d 26.4 ± 1.13bc
20 49.8 ± 0.37bcd 8.7 ± 1.87e 7.7 ± 0.33bc 83.8 ± 2.19e 29.5 ± 0.83 ab
30 51.4 ± 0.42 ab 9.6 ± 1.22de 7.4 ± 0.34c 82.4 ± 2.94e 30.3 ± 1.70a
40 49.7 ± 0.11bcd 9.8 ± 6.29de 7.2 ± 0.21cd 86.8 ± 4.23de 31.1 ± 0.26a
50 47.2 ± 0.68ef 11.4 ± 0.33de 6.8 ± 0.16d 83.9 ± 0.69e 30.6 ± 0.27a
LSD 1.81 7.3 0.51 7.54 3.18
CV 0.02 0.17 0.04 0.03 0.05

Each parameter was analysed five replicates (n = 5). The results were given as mean ± standard deviation.
Values followed by the different letters in the same column are significantly different (*p < 0.05; **p < 0.01).
SL: Substitution Level; RHBF: Refined Hull-less Barley Flour; WHBF: Whole grain Hull-less Barley Flour; nd: Not detected.
LSD: Least Significance Difference; CV: Coefficient of Variance.

Fig. 1. SDS-PAGE patterns of the total extractable proteins of DWS, WHBF, RHBF and uncooked spaghetti products in non-reducing (A) and reducing (with 5%
β-mercaptoethanol, B) conditions. Lane 1: DWS, Lane 2: Control spaghetti, Lane 3–7: 10–50% WHBF substituted spaghetti products, Lane 8: WHBF, Lane M: Wide-
range protein markers, Lane 9: RHBF, Lane 10–14: 10–50% RHBF substituted spaghetti products. MW: Molecular weight; HMW: High molecular weight.

enter the gel and they accumulated in the beginning of the resolving gel
(Fig. 1A). The HMW-glutenins at the molecular weight of 115 and 103
kDa could be clearly seen in the reducing conditions of the DWS and
control spaghetti (Fig. 1B, Lane 1–2). Their intensity decreased towards
to 50% hull-less barley flour (HBF) substitution levels (Fig. 1B, Lane 7
and 14). Visioli et al. (2018) also described the HMW- and low molecular
weight (LMW) glutenins between 80-140 kDa and 31–51 kDa, respec­
tively, each are joined by disulphide bonds. The sulphur-rich and
sulphur-poor prolamins of DWS (ω-, γ-, β-, α-gliadins) had molecular
weight between 28 and 60 kDa and their intensity decreased while
increasing of the HBF substitutions from 10% to 50% (Fig. 1A and
Fig. 1B, Lane 3–7 and Lane 10–14). Visioli et al. (2018) also indicated
the molecular weights of gliadins between 30 and 66 kDa in their study.
Gluten composition and the relative amounts of sub-units are known to
contribute to the technological quality of semolina (Visioli et al., 2018).
Barley hordeins are separated into four groups based on their elec­
trophoretic mobilities and amino acid compositions: the polymeric D
hordeins, monomeric C hordeins, monomeric γ hordeins, polymeric B
hordeins (Tatham et al., 2000). The main storage proteins of HBF were
D, C, γ and B hordeins and albumins and globulins (also called as A
hordeins) were indicated on Fig. 1B and their concentration increased in
the HBF substituted spaghetti samples while increasing the substitution
levels. The polymeric D hordeins could not be seen in the non-reducing
conditions of HBF substituted spaghetti samples (Fig. 1A, Lane 3–7 and
Lane 10–14) and HBF samples (Fig. 1A, Lane 8 and 9) as well.
Conversely, D hordeins could be seen at the molecular weight of 86 kDa
in the reduced condition of HBF substituted spaghetti samples (Fig. 1B,
Lane 3–7 and Lane 10–14) and also in the HBF samples (Fig. 1B, Lane 8
and 9, respectively). The monomeric C hordeins, which were much
visible on both electrophoregrams, were marked at the molecular
weight of 52 and 63 kDa in Fig. 1A, and they were marked at the mo­
lecular weight of 49 and 61 kDa in Fig. 1B. The samples in reducing
conditions showed that γ and B hordeins located at the molecular
weights between 29 and 45 kDa and there might be some polymerization
between barley and durum wheat proteins in this region and this was
more obvious towards the 50% substitution levels of both types of flours
(Fig. 1B, Lane 3–7 and Lane 10–14). Polymeric protein formation has
been described as one of the most important episodes initiated during
industrial pasta making in order to achieve high quality pasta (Padalino
et al., 2019).
4. Conclusions
The partial substitutions of RHBF and WHBF instead of DWS in
spaghetti were evaluated. The chemical, dietary fibre composition and
technological characteristics of HBF substituted spaghetti products were

7
D. Ermişer and E. Yalçın
compared to control spaghetti. This research demonstrated that RHBF or
WHBF could be incorporated into spaghetti. Both types of flours pro­
vided a concentrated source of dietary fibre in the forms of soluble and
insoluble fibres. An increasing of resistant starch (RS3) content after
cooking of spaghetti was first presented with HBF substitution in this
research. This study showed that ash, protein, total-, soluble- and
insoluble dietary fibres and β-glucan contents of spaghetti could be
increased with the HBF substitution and the nutritionally enriched
spaghetti products could be produced. Therefore, the partial substitution
of DWS with RHBF or WHBF could be an efficient way to increase the
soluble dietary fibre intake in human nutrition. The soluble and insol­
uble dietary fibres from HBF could affect the textural and cooking
properties of spaghetti. The results showed that up to 20% of HBF
substitution is promising for the production of spaghetti in industry. The
total extractable proteins of uncooked spaghetti products were investi­
gated on SDS-PAGE and the changes on main storage proteins of DWS
and hull-less barley due to HBF substitution level were first monitored.
Whole grain hull-less barley flour can be used to formulate health-
promoting spaghetti, however the modifications on formulation and
processing can be improved. The spaghetti produced with hull-less
barley flours can be a new value added product in the exporting cereal
based foods.
CRediT authorship contribution statement
Dönüş Ermişer: Project administration, Funding acquisition, Inves­
tigation, Methodology, Formal analysis, Statistics. Erkan Yalçın: Proj­
ect administration, Funding acquisition, Investigation, Methodology,
Formal analysis, Writing – original draft, Writing – review & editing.
Declaration of competing interest
The authors declare no conflict of interest.
Acknowledgement
This work was financially supported by the Republic of Turkey,
Ministry of Agriculture and Forestry, General Directorate of Agricultural
Research and Policies, Project # TAGEM/HSGYAD/17/A03/P01/122.
References
AACCI (American Association of Cereal Chemists International), 2010. Methods No: 08-
01.01, 14-50.01, 16-50.01, 38-12.02, 44-15.02, 46-30.01, 54-21.01, 55-10.01, 55-
31.01, 66.52.01. In: Approved Methods of Analysis, eleventh ed. The Association, St.
Paul, MN, USA.
Aalami, M., Prasado Rao, U.J.S., Leelavathi, K., 2007. Physicochemical and biochemical
characteristics of Indian durum wheat varieties: relationship to semolina milling and
spaghetti making quality. Food Chem. 102 (4), 993–1005.
Aravind, N., Sissons, M., Egan, N., Fellows, C.M., Blazek, J., Gilbert, E.P., 2012. Effect of
β-glucan on technological, sensory, and structural properties of durum wheat pasta.
Cereal Chem. 89 (2), 84–93.
Arndt, E.A., 2006. Whole-grain barley for today’s health and wellness needs. Cereal
Foods World 51 (1), 20–22.
Bourdon, I., Yokoyama, W., Davis, P., Hudson, C., Backus, R., Richter, D., Knuckles, B.,
Schneeman, B.O., 1999. Postprandial lipid, glucose, insulin, and cholecystokinin
responses in men fed barley pasta enriched with β-glucan. Am. J. Clin. Nutr. 69 (1),
55–63.
Journal of Cereal Science 102 (2021) 103315
Chillo, S., Ranawana, D.V., Henry, C.J.K., 2011. Effect of two barley β-glucan
concentrates on in vitro glycaemic impact and cooking quality of spaghetti. LWT-
Food Sci. Tech. 44, 940–948.
Ciccoritti, R., Nocente, F., Sgrulletta, D., Gazza, L., 2019. Cooking quality, biochemical
and technological characteristics of bran-enriched pasta obtained by a novel pasta-
making process. LWT - Food Sci. Technol. (Lebensmittel-Wissenschaft -Technol.)
101, 10–16.
Cleary, L., Brennan, C., 2006. The influence of a (1→3) (1→4)-β-D-glucan rich fraction
from barley on the physico-chemical properties and in vitro reducing sugars release
of durum wheat pasta. Int. J. Food Sci. Technol. 41, 910–918.
D’Egidio, M.G., De Stefanis, E., Fortini, S., Galterio, G., Nardi, S., Sgrulletta, D.,
Bozzini, A., 1982. Standardization of cooking quality analysis in macoroni and pasta
products. Cereal Foods World 27, 367–368.
De Paula, R., Abdel-Aal, E.-S.M., Messia, M.C., Rabalski, I., Marconi, E., 2017. Effect of
processing on the β-glucan physicochemical properties in barley and semolina pasta.
J. Cereal. Sci. 75, 124–131.
Fu, B.X., Sapirstein, H.D., 1996. Procedure for isolating monomeric proteins and
polymeric glutenin of wheat flour. Cereal Chem. 73, 143–152.
Gelencsér, T., Gál, V., Hódsági, M., Salgó, A., 2008. Evaluation of quality and
digestibility characteristics of resistant starch enriched pasta. Food Bioprocess
Technol. 1, 171–179.
Graziano, S., Marando, S., Prandi, B., Boukid, F., Marmiroli, N., Francia, E.,
Pecchioni, N., Sforza, S., Visioli, G., Gulli, M., 2019. Technological quality and
nutritional value of two durum wheat varieties depend on both genetic and
environmental factors. J. Agric. Food Chem. 67, 2384–2395.
ICC (International Association for Cereal Science and Techonology), 2002. Standard
Methods No.129.
Izydorczyk, M.S., Lagasse, S.L., Hatcher, D.W., Dexter, J.E., Rossnagel, B.G., 2005. The
enrichment of Asian noodles with fiber-rich fractions derived from roller milling of
hull-less barley. J. Sci. Food Agr. 85, 2094–2104.
Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of
Bacteriophage T 4. Nature 227, 680–685.
Lamacchia, C., Baiano, A., Lamparelli, S., Terracone, C., Trani, A., Di Luccia, A., 2011.
Polymeric protein formation during pasta-making with barley and semolina
mixtures, and prediction of its effect on cooking behaviour and acceptability. Food
Chem. 129, 319–328.
Makhlouf, S., Jones, S., Ye, S.-H., Sancho-Madriz, M., Burns-Whitmore, B., Li, Y.O., 2019.
Effect of selected dietary fibre sources and addition levels on physical and cooking
quality attributes of fibre-enhanced pasta. Food Qual. Saf. 3, 117–127.
Malcolmson, L., Lukie, C., Swallow, K., Sturzenegger, T., Han, J., 2014. Using barley
flour to formulate foods to meet health claims. Cereal Foods World 59 (5), 235–242.
Montalbano, A., Tesoriere, L., Diana, P., Barraja, P., Carbone, A., Spano, V., Parrino, B.,
Attanzio, A., Livrea, M.A., Cascioferro, S., Cirrincione, G., 2016. Quality
characteristics and in vitro digestibility study of barley flour enriched ditalini pasta.
LWT - Food Sci. Technol. (Lebensmittel-Wissenschaft -Technol.) 72, 223–228.
Ng, P.K.W., Bushuk, W., 1987. Glutenin of marquis wheat as a reference for estimating
molecular weights of glutenin subunits by sodium dodecyl sulfate-polyacrylamide
gel electrophoresis. Cereal Chem. 64, 324–327.
Padalino, L., Del Nobile, M.A., La Gatta, B., Rutigliano, M., Di Luccia, A., Conte, A., 2019.
Effects of microwave treatment of durum wheat kernels on quality characteristics of
flour and pasta. Food Chem. 283, 454–461.
Sinesio, F., Paoletti, F., D’Egidio, M.G., Moneta, E., Nardo, E., Peparaio, M.,
Comendador, F.J., 2008. Flavor and texture as critical sensory parameters of
consumer acceptance of barley pasta. Cereal Foods World 53 (4), 206–213.
Sullivan, P., Arendt, E., Gallagher, E., 2013. The increasing use of barley and barley by-
products in the production of healthier baked goods. Trends in Food Sci. Tech. 29,
124–134.
Tatham, A.S., Gilbert, S.M., Fido, R.J., Shewry, P.R., Marsh, Michael N., 2000.
Extraction, separation, and purification of wheat gluten proteins and related proteins
of barley, rye, and oats. In: Celiac Disease-Methods and Protocols. Humana Press,
Edt, pp. 55–73.
Tiwari, U., Cummins, E., 2009. Factors influencing β-Glucan levels and molecular weight
in cereal-based products. Cereal Chem. 86 (3), 290–301.
Visioli, G., Vamerali, T., Dal Cortivo, C., Trevisan, S., Simonato, B., Pasini, G., 2018.
Pasta-making properties of the new durum wheat variety Biensur suitable for the
Northern mediterranean environment. Ital. J. Food Sci. 30, 673–683.
Williams, P.C., El-Haramein, F.J., Nakkoul, H., Rihawi, S., 1986. Crop Quality Evaluation
Methods and Guidelines Technical Manual No: 14. International Center for
Agricultural Research in the Dry Areas, ICARDA, Aleppo, Syria, p. 145.
Yalçın, E., Çelik, S., Akar, T., Sayım, İ., Köksel, H., 2007. Effects of genotype and
environment on β-glucan and dietary fiber contents of hull-less barleys grown in
Turkey. Food Chem. 101, 171–176.