Environmental Technology

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Bioethanol production from pistachio (pistacia

vera L.) shells applying ozone pretreatment and
subsequent enzymatic hydrolysis

Betül Göncü, Hakki Gülşen & Emir Zafer Hoşgün

To cite this article: Betül Göncü, Hakki Gülşen & Emir Zafer Hoşgün (2021): Bioethanol production
from pistachio (pistacia�vera L.) shells applying ozone pretreatment and subsequent enzymatic
hydrolysis, Environmental Technology, DOI: 10.1080/09593330.2021.1903565

To link to this article: https://doi.org/10.1080/09593330.2021.1903565

Published online: 25 Mar 2021.

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ENVIRONMENTAL TECHNOLOGY
https://doi.org/10.1080/09593330.2021.1903565

Bioethanol production from pistachio (pistacia vera L.) shells applying ozone
pretreatment and subsequent enzymatic hydrolysis
a a b
Betül Göncü , Hakki Gülşen and Emir Zafer Hoşgün
a
Department of Environmental Engineering, Harran University, Şanlıurfa, Turkey; bDepartment of Chemical Engineering, Eskişehir Technical
University, Eskişehir, Turkey

ABSTRACT ARTICLE HISTORY

Pistachio (pistacia vera L.) is a lignocellulosic raw material. One of the most pistachio produced Received 7 January 2021
three countries in the World is Turkey and Şanlıurfa is the city that most pistachio production in Accepted 6 March 2021
Turkey. As a result of this production, a large amount of pistachio waste is generated. Therefore,
KEYWORDS
this study was conducted considering the abundant pistachio waste and furthermore, the Lignocellulosic biomass;
effects of ozone and combined (ozone and hot water) pretreatments for bioethanol production pistachio shells;
from pistachio shells were investigated. Initially, the ozone and combined pretreatments were fermentation; ozone
applied to the pistachio shells. It has been observed that applying the combined pretreatment pretreatment; bioethanol
provides better lignin removal than only ozone pretreatment and on the other hand, the ozone
pretreatment provides better lignin removal than the hot water pretreatment. Scanning
electron microscopy (SEM) images of pretreated and untreated pistachio shells were compared.
Enzyme activity was measured, and 30–60 FPU enzyme loading was applied in an enzymatic
hydrolysis. The enzymatic hydrolysis was applied to obtain fermentable sugar from the pistachio
shells after pretreatments. As a result of enzymatic hydrolysis, 2.34–8.24 g/L reducing sugar was
obtained. On the other hand, 1.21–2.33 g/L ethanol concentration was obtained end of the
fermentation process. Fermentation efficiency was calculated between 42% and 55%. As a
result, this study showed that combined pretreatment was more effective than the single
pretreatment in the ethanol production process.

Introduction fuels are not enough to meet this growing energy

demand. In addition to being non-renewable, fossil
Fossil fuels are the main source of energy used in the fuels are not sustainable and have many environ-
world [1]. The world’s energy demand is increasing by mental hazards, including greenhouse gas (GHG)
30% per year and the reserves of non-renewable fossil and carcinogenic exhaust emissions [2]. For these

CONTACT Betül Göncü betulgoncu@harran.edu.tr Department of Environmental Engineering, Faculty of Engineering, Harran University, Şanlıurfa,
Turkey
This article has been corrected with minor changes. These changes do not impact the academic content of the article.
© 2021 Informa UK Limited, trading as Taylor & Francis Group
2 B. GÖNCÜ ET AL.
reasons, the energy researches start to focus on the
renewable energy sources that can be used instead
of fossil fuels [3]. The increasing world population,
global warming, economic issues, and disadvantages
arising from the use of fossil fuels have encouraged
a roadmap for biofuel production in an environmen-
tally friendly manner [4,5]. Biofuels such as bioetha-
nol, biodiesel, biomethane and bio-oil are alternative
clean energy sources to the fossil fuels. Currently, bio-
fuels contribute ∼ 50 EJ to the world’s energy
demand [6].
Bioethanol is the most important biofuel, accounting
for ∼65% of the world’s biofuel production to date [7].
Basically, bioethanol has a high octane number, so it
can be used both as an octane enhancer and a fuel in
gasoline-powered vehicles [8]. Bioethanol can be pro-
duced from inedible sources such as agro-industrial
and wood-cellulosic waste [9]. Most of bioethanol pro-
duced is obtained from food products such as corn,
sugar beet and potatoes [10]. The use of food raw
materials and agricultural land to obtain biofuels has
increased the food prices and led to a much larger
problem in developing countries, namely the food
crisis [11]. Therefore, lignocellulosic raw material
obtained from agricultural, industrial and forestry resi-
dues has become an important source for the biofuel
production by making a high contribution to the
concept of circular economy and is saved from being
waste [12]. The main components of lignocellulosic
biomass are cellulose, hemicellulose and lignin, which
provides structural integrity [13]. Lignocellulosic raw
materials are highly resistant to enzymatic and biological
processes; therefore, a pretreatment process is required
to break down the complex structure and release cellu-
lose fibres for hydrolysis and fermentation [14]. Physical,
chemical and biological pretreatment methods or a
combination of two or more pretreatment processes
can be used to produce bioethanol from the lignocellu-
losic materials [15]. However, bad reaction conditions
and eliminate the possibility of inhibitor formation can
lead to high costs in pretreatment processes. Hence,
using the minimum amount of energy and chemicals,
preventing the formation of degradation products and
toxic substances during the process will reduce pretreat-
ment costs [16]. The hydrolysis process is carried out
after the pretreatment stage. The conversion of lignocel-
lulosic raw material to monomeric sugar by hydrolysis is
currently the most challenging stage in bioethanol pro-
duction. The difficulty in the hydrolysis phase of ligno-
cellulosic biomass is that it is desired to obtain a
higher amount of alcohol at a lower investment cost
by reducing the use of chemicals and energy in the pre-
treatment process [17].
Ozone and hot water pretreatments applied to ligno-
cellulosic wastes were used in this study. The main
advantage of the ozone is that it is a powerful oxidant
and can dissolve easily in water [18]. The ozone pretreat-
ment causes a high amount of lignin removal by increas-
ing the reaction rate of ozone. It also does not create
toxic materials and can be carried out at room tempera-
ture, which is a significant advantage as it reduces the
pretreatment cost [19]. Since the hot water pretreatment
does not require the use of expensive chemicals, it
reduces the cost of pretreatment and does not create
environmental pollution. In this respect, it is an impor-
tant alternative method [20].
Season 2019/2020 was an ‘off year’ for US and Turkish
pistachios, while Iran experienced an ‘on year’ after over-
coming the short crop of the prior season. Consequently,
world production added up to 655,200 metric tons
(inshell basis), 15% down from the previous year, yet
12% above 2017/18, the last predominantly offseason.
The U.S.A. and Iran led global production, accounting
for 51% and 31% of the world share, respectively.
Along with Turkey, they represent 93% of the pistachio
world supply [21]. Recent statistics showed that one of
the three countries with the highest pistachio pro-
duction in the world is Turkey as 2,40,000 tons. Şanlıurfa
is the city where highest pistachio production with
1,00,107 tons, and also 55,686 tons of pistachio waste
is occurred in Turkey and it shows that 42% of Turkey
pistachio production made in Şanlıurfa [22,23]. Free
and abundant raw material, easy supply, sustainable,
minimum transport cost, etc. are reasons for using pista-
chio shell in this study in Şanlıurfa. In addition, there is
no study on the bioethanol production of the pistachio
shell. All these reasons inspired bioethanol production
using the pistachio shell in this study.
Materials and methods
Materials
Cellic Ctec2, an enzyme mixture of cellulase, hemicellu-
lase and ß-glucosidase, was used. All chemicals and
enzymes used in the study were procured from Sigma-
Aldrich (U.S.A.).
Methods
Pistachio shell was collected in Şanlıurfa province,
Turkey. It was dried at 105°C before using in the study
and was grinded in a mill to reduce particle size. A par-
ticle size smaller than 2 mm was used. It was stored at
room temperature in a sealed glass bottle. The compo-
sition of feedstock (lignin, cellulose, and hemicellulose)

was determined using NREL methods. During this study,
analyses were repeated two times and an average value
was used. Lignin, cellulose, and hemicellulose content of
the pistachio shell were determined as 32.34%, 20.74%
and 36.92% on a dry basis, respectively.
In literature, Hesam et al. determined on a dry basis
that lignin, cellulose, hemicellulose, and ash contents
of pistachio shell as 20.77 ± 1.10%, 31.13 ± 0.42%,
42.27 ± 1.62% and 0.89 ± 0.02%, respectively [24]. On
the other hand, Çelik et al. determined on a dry basis
that lignin, cellulose, hemicellulose, and ash contents
of pistachio shell as 37%, 21.41%, 14.31% and 3.52%,
respectively [25]. It is thought that the content of the pis-
tachio shell differs due to different environment con-
ditions in which raw material is grown, harvest time
and analytical method.
Pretreatments
Pistachio shells were pretreated using an ozone genera-
tor (Ozoneks, TR) that produces ozone using oxygen
from the air. Four pretreatment times (5, 15, 30 and
60 min), 8 mg/min ozone dosage concentration and 1/
10 solid/liquid ratio (S/L) were applied at room tempera-
ture. During the pretreatment period, there was no need
for extra mixing as ozone gas would form bubbles in the
water and provide mixing by itself. After pretreatment,
the samples were filtered. The solid fraction was col-
lected and dried for use in the lignin and enzymatic
hydrolysis step.
Ozone and liquid hot water sequential pretreatments
were with an ozone generator followed by the hot water
pretreatment in an autoclave. First, pistachio shells, 1/10
S/L ratio in a screwed borosilicate glass bottle, were
treated with 8 mg/min ozone concentration at a room
temperature for 5, 15, 30 and 60 min, respectively. After-
wards, the bottles were closed, and pretreatment of hot
water was applied at 100°C in the autoclave for 5, 15, 30
and 60 min, respectively. The effects of these two
different combined pretreatments on the lignin
removal, reducing sugar concentration and bioethanol
production compared to single ozone pretreatment
were investigated. As a result of these pretreatments,
the samples were filtered. The solid fractions were col-
lected and were dried using lignin analyses and enzy-
matic hydrolysis process.
Determination of lignin amount
Lignin content of the pistachio shell was determined
using a NREL method. First, gooch crucible was dried
for a minimum of six hours at 105°C and was put in a
desiccator. After cooling the crucible was weighed out.
ENVIRONMENTAL TECHNOLOGY 3
0.3 g (c1) of pistachio shell and 3 mL H2SO4 (72%) sol-
ution was added to the bottle. The bottle was stirred
for one minute and incubated at 30°C for 60 min.
About 84 mL water was added for diluting the solution
to 4% and the bottle was autoclaved at 121°C for
60 min. The solution was filtered with a crucible after
the solution has cooled down. The crucible was dried
for a minimum of 6 h at 105°C and was put in a desicca-
tor. The crucible was weighed out after it has cooled
down (c2). At the end of these processes, the crucible
was put in a furnace for minimum 24 h at 600°C. The cru-
cible was taken from the furnace and was weighed out
(c3) [26]. Percentage of lignin was calculated according
to Equation (1) [27]:


c2 − c3
Lignin(%) = × 100. (1)
c1
Delignification was calculated according to Equation
(2) [27]:
Delignification(%) =
⎛Lignin of untreated biomass ⎞
⎜ − Lignin of pretreated biomass⎟
⎝ ⎠ × 100.
Lignin of untreated biomass
(2)
Scanning electron microscopy (SEM)
Zeiss Evo 50 system SEM was used to observe the surface
morphology of the pistachio shells. This device that
including EDX spectrometer was set at 20 kV to use
the SEM analysis. A maximum delignification condition
was selected to observe on the pistachio shell.
Measurement of enzyme activities and enzymatic
hydrolysis
The NREL (NREL/TP-510-42628) method was used for the
determination of enzyme activity. The enzyme activity of
Cellic CTec2 which is used in the enzymatic hydrolysis
was determined. About 1416 mL of deionized water,
10.6 g of 3,5-dinitrosalicylic acid (C7H4N2O7), 19.8 g of
sodium hydroxide (NaOH), 306 g of potassium sodium
tartrate (KNaC4H4O6·4H2O), 7.6 mL of phenol (C6H5OH)
and 8.3 g of sodium metabisulphite (Na2S2O5) were
added to prepare DNS (dinitrosalicylic acid) solution.
10 mg/mL glucose stock solution and a citrate buffer sol-
ution were used to prepare four different glucose
dilution solutions. Glucose solutions at different concen-
trations were prepared by adding 0.5, 1, 2 and 4 mL of
buffer solution separately to 1 mL of glucose stock
4 B. GÖNCÜ ET AL.
solution in different tubes. Then 0.5 mL of each solution
was taken, and 1 mL of buffer solution was added to
them. A 1.5 mL citrate buffer solution was added to
the control tube, 0.5 mL of the enzyme solution was pre-
pared at different concentrations and 1 mL of the buffer
solution was added to each one. Different concen-
trations of 0.5 mL enzyme solution, 1 mL of buffer sol-
ution and 50 mg of Whatman No. 1 filter paper were
placed in enzyme control tubes. All glucose standard
tubes, control tubes and enzyme determination tubes
were incubated at 50°C for 60 min. Then 3 mL of DNS
solution was added to all tubes and were boiled for
exactly 5 min and suddenly cooled in cold water bath.
UV–vis Spectrophotometer measurement were carried
out at 540 nm. The glucose solutions were diluted by
adjusting the absorbance values to be between 0.1
and 1. A glucose calibration curve was obtained. The
amount of glucose in the enzyme tubes was calculated
after subtracting the absorbance in the control tube
from each enzyme ratio absorbance using the cali-
bration curve. The enzyme concentration that will
produce 2 mg of glucose was determined. A filter
paper unit was determined according to the amount
of enzyme that produced 2 mg of reducing sugar in
60min from 50 mg of filter paper [26,27].
Enzymatic hydrolysis processes were carried out
using a citrate buffer and an enzyme to ozone and com-
g 
bined (ozone + hot water) pretreated pistachio shells. 30
and 60 FPU enzyme were applied for the processes.
About 5% (w/v) substrate was loaded using the citrate
buffer. An orbital shaker incubator was used to keep
the temperature of reactors at 50°C, for 72 h [28,29].
Reducing sugar concentration
Dinitrosalicylic acid (DNS) method was used to deter-
mine reducing sugar concentration. About 1 mL 0.05 M
of buffer solution (pH = 4.8) and 0.5 mL hydrolyzate
were put into each tube. Blank was selected as 1.5 mL
buffer solution. Incubator was used to keep the temp-
erature of tubes at 50°C for 60 min. 3 mL DNS was
added to the tubes and boiled for 5 min. Then suddenly
cooled in an ice bath and absorbance was measured at
540 nm. The amount of reducing sugar was calculated
using the absorbance values [30,31].
Fermentation of pistachio shells
The hydrolyzates obtained end of the enzymatic
hydrolysis were filtered and were centrifuged (4°C,
10000 rpm, 5 min) to remove the unhydrolyzed
residue. Saccharomyces cerevisiae, the most widely
used and cheapest yeast, was used as a microorganism.
Yeast growth was carried out before fermentation. 3 g/L
yeast extract, 3.5 g/L peptone, 1 g/L ammonium sul-
phate ((NH4)2SO4), 0.5 g/L calcium chloride (CaCl2), 2 g/
L potassium phosphate (KH2PO4), 1 g/L magnesium sul-
phate tetrahydrate (MgSO4·7H2O), were used for it. The
prepared solution and glasswares were sterilized at
121°C in autoclave for 15 min. The yeast was growth at
30°C and 100 rpm for 24 h. 10% (v/v) S. cerevisiae and
the hydrolyzate were mixed and fermentation was
carried out at pH 4.8 for 48 h. HPLC (Agilent 1100,
Germany) was used to determine ethanol concentration.
Refractive index detector and Bio-Rad Aminex HPX-87H
(U.S.A.) column (300 mm × 7.8 mm) were used in a
HPLC system. The column was operated at 60°C,
0.005M H2SO4 solution was used as the mobile phase
at a flow rate of 0.6 mL/min. The fermentation
efficiency was calculated using Equation (3) [32]:
Fermentation efficiency (%)


ethanol produced
= × 100 . (3)
theoritic ethanol
Theoretical ethanol assumes that all initial glucose is
converted to ethanol during fermentation and that
conversion results in 0.51 g ethanol/g glucose. The
productivity of ethanol was determined to using
Equation (4) [32]:
Ethanol productivity
L×h


production of ethanol (g/L)
= . (4)
time (h)
Results and discussion
Pretreatments
Ozone pretreatment applied that 1/10 (w/v) solid/liquid
ratio, pretreatment time of 5, 15, 30 and 60 min and at
room temperature, for delignification of the pistachio
shells. Applying ozone pretreatment was showed that
the delignification yield was ranged from 23.28% to
37.43%. Combined (ozone + hot water) pretreatment
applied that 1/10 (w/v) solid/liquid ratio, pretreatment
time of 5 min ozone + 5 min hot water, 15 min ozone
+ 15 min hot water, 30 min ozone + 30 min hot water,
60 min ozone + 60 min hot water, for the delignification
of the pistachio shells. Hot water pretreatment was
applied at 100°C, although ozone pretreatment was
applied at room temperature for combined pretreat-
ment. Applying combined pretreatment showed that
the delignification yield ranged from 26.22% to
41.22%. These results showed that maximum lignin
removal obtained by 60 min ozone + 60 min hot water

pretreatment and combined pretreatment provided
approximately 4% more lignin removal than ozone pre-
treatment. The delignification of ozone and combined
pretreatments are shown in Figure 1.
Garcia-Cubero et al. studied ozone pretreatment at
ozonation time of 2.5 h and reported that the
maximum lignin removal as 44% for grain straw [33].
Ortega et al. studied ozone pretreatment at an ozona-
tion time of 60 min and reported that the maximum
lignin removal as 47% for sugarcane straw [34].
Wobiwo et al. studied the hot water pretreatment
between 100°C and 120°C and reported that the
maximum lignin removal as 16% for banana bulb [20].
Xia et al. studied the hot water pretreatment between
130°C and 170°C and reported that the maximum
lignin removal was 10% for reed [35]. These studies
showed that lignin removal by ozone pretreatment
may differ depending on the lignin content of the raw
material and the pretreatment time. On the other
hand, hot water pretreatment has no strong effect on
lignin removal.
Figure 1. Effect of ozone and combined pretreatment (1/10 S/L).
ENVIRONMENTAL TECHNOLOGY 5
As seen in this study that the ozone pretreatment has
a high lignin removal rate. Conversely, hot water pre-
treatment provided low lignin removal in this study.
However, as can be seen from other studies, the
applied temperature in hot water pretreatment is an
important factor. For this reason, it is foreseed that
better lignin removal can be obtained if a higher temp-
erature is applied for the hot water pretreatment.
Looking at the combined pretreatment, it was seen
that it provided better lignin removal than single
pretreatment.
SEM analysis
Morphological differences of untreated and ozone hot
water pretreated the pistachio shell was assessed by
using SEM. The pistachio shell was coated with gold. In
addition, an SEM image was used to observe the struc-
tural change that occurred after pretreatment. SEM
images are presented in Figure 2. The first image
Figure 2. SEM images (a) untreated and (b) pretreated.
6 B. GÖNCÜ ET AL.
showed that fibrils have a continuous structure. In case
pretreatment was applied, the structures were signifi-
cantly deformed or fragmented. Depolymerization of
lignin after ozone and hot water pretreatment can be
seen by the conversion of long structured fibres into
short and closed individual fibres.
Enzymatic hydrolysis
Pistachio shells were subjected to enzymatic hydrolysis
to produce reducing sugar. All enzymatic hydrolysis
process was realized 5% (w/v) substrate loading, using
citrate buffer, 50°C temperature, 30 or 60 FPU enzyme
loading per 1 g substrate, for 72 h. Enzyme activity of
Cellic Ctec2 was measured as 308 FPU/mL. Figure 3
shows the results of 30 FPU of enzyme loading of
Figure 3. Enzymatic hydrolysis of ozone and combined pre-
treated pistachio shells (30 FPU).
ozone and combined pretreated pistachio shell.
Maximum reducing sugar obtained from enzymatic
hydrolysis of ozone pretreated pistachio shell and com-
bined pretreated pistachio shell was 5.83 and 6.48 g/L
for 30 FPU enzyme loading, respectively.
Figure 4 shows the obtained reducing sugar concen-
tration after enzymatic hydrolysis (60 FPU per 1 g
enzyme loading) of the ozone and combined pretreated
pistachio shell. Maximum reducing sugar was obtained
as 7.31 and 8.24 g/L from enzymatic hydrolysis of
ozone pretreated pistachio shell and combined pre-
treated pistachio shell for 60 FPU enzyme loading,
respectively. As a result, combined pretreatment led to
an increase in reducing sugar obtained after enzymatic
hydrolysis for 30 and 60 FPU.
Subhedar et al. applied alkali and ultrasonic-assisted
alkali pretreatment to pistachio shell and obtained 8.1,
18.4 g/L reducing sugar, respectively [36]. In another
Figure 4. Enzymatic hydrolysis of ozone and combined pre-
treated pistachio shells (60 FPU).

study, Göncü et al. applied alkali pretreatment to pista-
chio shell and obtained 6.98 g/L reducing sugar [27].
These results show that the combined pretreatment is
more effective to produce reducing sugar than single
pretreatment.
Fermentation of pistachio shells
Enzymatic hydrolysis process was applied to pistachio
shells for 72 h after ozone and combined pretreatment.
End of these processes, hydrolysates were obtained to
use in fermentation. Fermentation was carried out
using S. cerevisiae yeast at 30°C for 48 h to obtain
bioethanol from hydrolyzates. Ethanol results were
showed in Figure 5. Ethanol concentration was
Figure 5. Ethanol results of ozone and combined pretreated pis-
tachio shells (60 FPU).
ENVIRONMENTAL TECHNOLOGY 7
Figure 6. Fermentation efficiency of ozone and combined pre-
treated pistachio shells (60 FPU).
calculated as 1.21, 1.36, 1.54 and 2.08 g/L, and ethanol
productivity was calculated as 0.05, 0.057, 0.064 and
0.086 from 5, 15, 30 and 60 min ozone pretreated hydro-
lyzate after fermentation, respectively. On the other
hand, 1.32, 1.49, 1.71 and 2.33 g/L ethanol was obtained,
and ethanol productivity was calculated as 0.055, 0.062,
0.071 and 0.097 from 10, 30, 60 and 120 min combined
pretreated hydrolyzate, respectively. Fermentation
efficiencies were showed in Figure 6. The fermentation
efficiency of ozone pretreated hydrolyzate was calcu-
lated between 42% and 52% and furthermore, fermenta-
tion efficiency of combined pretreated hydrolyzate was
calculated between 44% and 55%.
Conclusions
In this study, bioethanol production from the pistachio
shell applying ozone and combined (ozone + hot
water) pretreatment was investigated. Ozone pretreat-
ment was applied at 5, 15, 30, 60 min and combined pre-
treatment was applied at 5 + 5, 15 + 15, 30 + 30 and 60 +
60 min. The maximum lignin removal was obtained as
8 B. GÖNCÜ ET AL.
37.43% by ozone pretreatments and this was improved
to 41.22% by combined pretreatments. When ozone
and hot water pretreatment was applied sequentially,
it was observed that there was an increase in efficiency
in lignin removal compared to single ozone pretreat-
ment. All the pretreatment conditions were applied to
the pistachio shell to obtain maximum reducing sugar
after enzymatic hydrolysis. Enzymatic hydrolysis pro-
cesses were applied with 30 and 60 FPU enzyme
loading. Enzymatic hydrolysis of pistachio shells with
30 FPU enzyme loading, maximum reducing sugars
were obtained 5.83 and 6.48 g/L in ozone and combined
pretreatment, respectively. Further increase in the
enzyme loading from 30 to 60 FPU led to increase in
the maximum reducing sugar yields to 7.31 and
8.24 g/L in ozone and combined pretreatment, respect-
ively. The fermentation process was applied to hydroly-
zates obtained after enzymatic hydrolysis of 72 h.
Maximum ethanol yield was obtained 2.33 g/L from
combined pretreated pistachio shells.
As a result of this study, it was observed that com-
bined pretreatment increased reducing sugar concen-
tration and the delignification compared to single
pretreatment. Hence, the amount of produced bioetha-
nol from the combined pretreatment was also higher
than the single pretreatment. Different sequential pre-
treatments may be performed to the pistachio shell to
increase the delignification and thus obtained more
bioethanol in future works.
Disclosure statement
No potential conflict of interest was reported by the author(s).
ORCID
Betül Göncü http://orcid.org/0000-0003-0175-8078
Hakki Gülşen http://orcid.org/0000-0002-0726-555X
Emir Zafer Hoşgün http://orcid.org/0000-0002-3810-701X
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