Arabian Journal for Science and Engineering (2022) 47:15705–15725

https://doi.org/10.1007/s13369-022-06659-y

RESEARCH ARTICLE-MECHANICAL ENGINEERING

Optimization of Electricity Generation Parameters with Microbial Fuel

Cell Using the Response Surface Method
Süleyman Neşeli1 · Kevser Dincer2 · Şakir Taşdemir3 · Mustafa Akram Hayder Hayder2

Received: 1 July 2021 / Accepted: 30 January 2022 / Published online: 26 March 2022
© King Fahd University of Petroleum & Minerals 2022

Abstract
Due to an ever-increasing population and developments in technology, the demand for energy has been increasing daily. In
order to meet this demand, renewable and alternative energy sources that do not harm the environment are often recommended.
One of these alternative sources can be obtained from human wastewater and is known as the Microbial Fuel Cell (MFC).
This study aimed to estimate the electricity production performance of MFC, using Nafion 117 membrane with 10 × 10 and
11 × 11 cm2 area by both active and sediment sludges. The responses obtained from 16 physical experiments performed
according to the L 16 (24 × 22 ) Taguchi orthogonal index were analyzed by the response surface method (RSM). According
to the analysis, it was determined that the quadratic polynomial equations created for the estimation of the reaction values of
the active and sediment sludge had validity over 90%. The ANOVA analysis applied to determine parameter ethicalities also
showed that the most effective parameter on the reactions from the sludge types considered was the resistance at different
rates. Thus, the results suggest that the orthogonal design-based RSM model is an effective tool that provides key information
for optimizing energy harvesting from MFC technology and saves time concerning experimental work.

Keywords Microbial fuel cells · Response surface methodology · Orthogonal design · Active and sediment sludge

1 Introduction these resources vary regionally. In addition to popular alter-

native energy sources, Microbial Fuel Cells (MFCs) come
Technology is advancing rapidly in today’s world. Conse- to the forefront as an environmentally friendly but non-
quently, the need for energy is also increasing. Although regional energy source used in electrical energy production
fossil fuels have been the primary source of inexpensive by making use of organic materials in wastewater, an ele-
energy to power industrial applications, they are harmful to ment found in every environment in which people live.
the environment due to the waste they produce and have a lim- MFCs are bio-devices that can recover the energy stored in
ited amount for use. Therefore, researchers have been looking organic bonds in the form of electrical energy thanks to the
for a promising renewable energy source as an alternative metabolism of microorganisms by simultaneously removing
to fossil fuels [1, 2]. Among the available energy sources, organic waste [3–9]. The working principle of an MFC is
wind, sea currents, and solar energy are the most used alterna- presented in Fig. 1. An MFC converts energy, available in
tive and environmentally friendly energy sources. However, a bio-convertible substrate, directly into electricity. This can
be achieved when bacteria switch from the natural electron
acceptor, such as oxygen or nitrate, to an insoluble accep-
tor, such as the MFC anode. This transfer can occur either
B Süleyman Neşeli
sneseli@selcuk.edu.tr via membrane-associated components, or soluble electron
shuttles. The electrons then flow through a resistor to a cath-
1 Mechanical Engineering Department, Faculty of Technology, ode, at which the electron acceptor is reduced. In contrast
Selcuk University, Selçuk, Turkey
to anaerobic digestion, an MFC leads to electrical current
2 Mechanical Engineering Department, Faculty of Engineering and an off-gas containing mainly carbon dioxide [10, 11].
and Natural Science, Konya Teknik University, Konya,
To best understand how MFCs produce electricity, it is first
Turkey
necessary to state how bacteria absorb and process energy.
3 Computer Engineering Department, Faculty of Technology,
Selcuk University, Selçuk, Turkey

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different wastewaters in MFCs: potato wastewater and ani-

mal wastewater. Both types generated power in MFCs, with
the potato wastewater generating 217 mW/m2 of power
and the animal wastewater 189 mW/m2 . Xin et al. [20]
investigated electricity generation and the microbial com-
munity of single-chamber microbial fuel cells in response
to Cu2 O nanoparticles/reduced graphene oxide as a cathode
catalyst. They noted that the Cu2 O nanoparticles decorated
reduced graphene oxide (Cu2 O/rGO) as the cathode catalyst
of simple-chamber microbial fuel cells could produce higher
the maximum output voltage and coulombic efficiency when
compared with a commercial Pt/C (20 wt%) catalyst. The
results indicated that an MFC with a Cu2 O/rGO cathode cata-
lyst produced higher output voltage (0.223 V) and coulombic
efficiency (92.5%) when compared with a commercial Pt/C
(0.206 V, 90.3%). Mohan et al. [21] introduced critical fac-
tors regulating the bio-catalyzed electrochemical processes
Fig. 1 The working principle of an MFC and recent advancements in MFCs.
The effects of these parameters can determine the bio-
logical activity of microorganism and the type of current
Bacteria transform and store energy into the form of adeno- extraction [22]. Recently suggested numerical and statistical
sine triphosphate (ATP) by catheterizing chemical reactions. models have considered biological, chemical, and opera-
In some bacteria, the reduced substrates are oxidized and tional factors [23, 24]. When these factors are coupled with
transferred to respiratory enzymes with NADH, the reduced the geometry and architecture of the MFC reactors [25], and
form of nicotinamide adenine dinucleotide (NAD). These the response and performance of MFCs can be tuned and
electrons flow through a series of enzymes that function for regulated.
the passage of respiratory chain enzymes into the inner mem- As an efficient and practical approach to determine the
brane and form a proton gradient. The protons are sent back effects of these parameters, the response surface methodol-
into the cell with the enzyme ADP, and 1 ADP molecule ogy (RSM) can be proposed. The use of RSM has already
is created from 1 adenosine diosate molecule for every 3–4 been applied in some biotechnology processes [26–29] as
protons. Electrons are finally delivered to the soluble end well as in MFCs and experimental modeling and design using
electron acceptor, such as nitrate, sulfate, or oxygen [12, 13]. the RSM have been used for maximizing power output [30].
Electric current generation is made possible by keeping Despite this, more studies are necessary to consolidate this
microbes separated from oxygen or any other end terminal method to enhance the understanding of empirical phenom-
acceptor other than the anode, and this requires an anaerobic ena involved in MFCs and to predict and manipulate the
anodic chamber. Typical electrode reactions are shown in performance of MFCs [31–36]. These kinds of studies can
Eqs. (1) and (2) using acetate as an example substrate [14]. contribute to increasing the market share of this technology
and to realizing scaled-up systems [37]. Thus, some of the
Anodic reaction : CH3 COO− + 2H2 O results reported in the literature are compared in Table 1.
This study aimed to estimate the electricity production
→microbes 2CO2 + 7H+ + 8e− (1)
performance of MFCs, using Nafion 117 membrane with 10
× 10 and 11 × 11 cm2 area by both active and sediment
Cathodic reaction : O2 + 4e− + 4H+ → 2H2 O (2) sludges. For this purpose, the response surface methodology
(RSM) was used. As RSM involves a Taguchi orthogonal
There are many factors that affect the performance of design, quadratic regression analysis was used to develop
MFCs. The most important of which are the genus of bac- empirical models for the response parameters. Thus, the
teria, bacterial metabolism, electron transport, membrane parameter combination to be created for the most econom-
performance, the internal resistance of electrolytes, temper- ical electricity generation was estimated. At the end of this
ature, and geometric structure of the MFC [15–17]. Kim study, the authors examined a novel approach for electricity
et al. [18] investigated challenges in microbial fuel cell produce prediction using MFCs. This research will be benefi-
development and operation and identified the main limit- cial for future real-world applications such as manufacturing
ing factors in MFC operation, providing recommendations plants.
to improve performance. Patrick et al. [19] examined two

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 Table 1 Additional literature survey on MFC studies

References Voltage Rout Current/Current Peak Power/Power Remarks

density density

This study (For 0.340 V 500  0.227 A/m3 77.067 W/m3 According to the analyses conducted within this study, the quadratic polynomial
active sludge) equations created for the estimation of the reaction values of the active and sediment
sludge have validities over 90%. The ANOVA analyses applied to determine
parameter ethicalities also determined that the most effective parameter on the
reactions from the sludge types considered was the resistance at different rates
This study (For 0.310 V 500  0.213 A/m3 71.333 W/m3
sediment sludge)
38 4.5 A/m2 for 42 A/m2 for 28 mL In MFCs, increasing the cathode specific surface area is the most important factor for
28-mL wastewater. The cell voltage as well as the volumetric power density increases
39 0.15 0.5 k − 44 mA/m2 − 14 mW/m2 The impact of applied external resistance during the formation of anodic bacterial
communities and wastewater to MFCs had no significant effect on the power
performance of MFCs
40 205 mV 470  − 40 mW/m2 Maximum voltage decreased in proportion to the acetate concentration
Arabian Journal for Science and Engineering (2022) 47:15705–15725

41 1k 0.2 mA Current generation increased slightly with cyanide at low concentrations (up to
1.5 mM)
42 − 820 (mA/m2 ) ∼480 mW/m2 for The power transformation of the response revealed that the transformed response no
pH  5.80 longer suffered from lack of fit. P-value of less than 0.0001 implied that the
transformed response for power density was highly significant
43 − 240 mV − 17 mA − 36 mW/m2 for Although higher buffer concentrations improves power production and COD reduction
optimization test in MFC as a result of higher conductivity and constant pH, very high buffer
concentrations might lead to power density reduction because of precipitation of salt
on electrodes and membranes
44 58.19 mW/m2 At low and medium levels of aeration, the power density was low. In addition, when Pt
loading was less than 0.3 mg/m2 , the amount of power density produced was not
significant
45 400–600 mV 1000  0.4–0.6 mA 888.9 ± 10.5 Both power outputs and total sulfate removal efficiencies increased first and then
mW/m2 decreased with the increase of COD/sulfate ratio and HRT, respectively. RSM was
then performed to optimize the system, with the target of maximizing power outputs
and total sulfate removal efficiencies, respectively
46 − 25 12.88 (mW/m3 ) The power generation capacity of the MFC increases with an increased concentration
of COD, but concentration beyond 30.000 mg/L showed a downward trend in power
generation
47 1.233 V − 1100 mA/m2 374.4 mW/m2 In terms of high glucose concentration and low G/Y ratios, when the glucose/yeast
ratio is very low, the OCV of mediatorless yeast-MFCs had a high value of 0.7 V and
then slightly decreased to 0.6 V as the glucose concentration increased

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Table 2 Properties of active and

sediment sludge [39] Active Sediment

Range Typical value Range Typical value

Total solid matter (%) 0.4–1.5 1 2.0–7.0 5.0

Volatile solid matter (%) 60–80 75 60–80 65
Specific weight 1.01 1.02
Solvable in ether (%) 5.0–12 6.0–30
Extract in ether (%) 7.0–35
Protein (%) 32–41 20–30 25
Azote (N, %) 2.4–5.0 1.5–4.0 2.5
Phosphorus (P2O5, %) 2.8–11.0 0.8–2.8 1.6
Potassium oxide (K2O, %) 0.5–0.7 7 0–1.0 0.4
Cellulose (%) 7 9.0–13 10
Iron (not sulfur, %) 2.0–4.0 2.5
Silica (SiO2, %) 8 15–20
ph 6.5–8.0 7 5.0–8.0 6.0
Alkalinity (mg/L CaCO3) 580–1100 500–1500 600
Organic acid (mg Hac/L) 1100–1700 200–2000 500
Energy content: kJ/Kg 23,300 23,300 18,600
Btu/lb 10,000 10,000 8000

Fig. 2 Schematic diagram of the experimental system of MFC a section of anode and cathode b Measuring system

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Fig. 3 a Copper weave (for anode) b Copper rod (for cathode)

2 Materials and Methods

In this study, active and sediment sludges were used for

electricity generation with MFCs. Active sludge is the most
widely used biological wastewater treatment process. As a
system, it is based on aerobic, mesophilic, and hanging cul-
ture and contains different types of microorganisms. The
types of microorganisms in active sludge also vary depend-
ing on the type of organic matter, its concentration, and
environmental factors (such as pH, temperature, oxygen con-
centration and a toxic substance) [13, 38]. Sediment sludge
is formed by separating inorganic and organic suspended
solids from the water by gravity. Sediments in wastewater
are defined as sediment sludge. Since sediment sludges are
rich in organic matter, stabilization should be done after the
condensation process [13]. The general characteristics of the Fig. 4 Modeling and optimization process with RSM
active and sediment sludge are given in Table 2.
placed in the anode section and a copper rod was placed in
the cathode section. Airflow velocity was measured with a
2.1 MFC configuration and measurements rotameter [13] Fig. 3.
For each sample in the experiment, the output parame-
The physical and schematic view of the experimental setup
ters were performed using different resistances, temperature
created to obtain electricity from active and sediment sludges
changes, volumetric flow rates, and area input parameters
is presented in Fig. 2.
were obtained by measuring current, voltage, and power. Cur-
(Fig. 2.a and b), the system is marked from 1 to 10,
rent density and power density were calculated using Eqs. (3)
Temperature sensor (thermal couple) (1), #ASMFC’s glass
and (4) equations, respectively.
containers (2), Copper braid (anode) and copper rod (3), The
magnetic stirrer heater (4), The table (5), The flow meter
(6), Multimeters (7), 10 × 10 and 11 × 11 cm2 Nafion I
Current density  (3)
membrane (8), External resistance-Potentiometer (Rout ) (9), V
Air motors (10).
In this study, an MFC was produced consisting of two I 2 x R0
Power density  (4)
glass chambers of 15 cm × 15 cm × 15 cm3 in dimension. V
Water was placed in the cathode section of the chambers and
sludge in the anode. 10 × 10 and 11 × 11 cm2 Nafion 117 In Eqs. (3) and (4), I is current, V is volume, and Ro is
membranes were placed in the middle of the glass chambers. outer resistance.
Due to its high ionic conductivity (10–2 S.cm−1 ), Nafion The procedure for the measurements performed in the
membrane was preferred. Because of copper’s ability to react experimental setup given in Fig. 2 and its related calcula-
and supply good electrical conductivity, copper weave was tions are listed below [13]:

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Fig. 5 Flow diagram of RSM

Table 3 The defined values for

each level of studied parameters Symbol Parameter Unit Level 1 Level 2 Level 3 Level 4

R Resistance Ohm 2 500 900 10,000

ΔT Temperature change °C 8 10 12 14
V̇ Volumetric flow rate L/h 1 1.5 – –
A Area cm2 100 121 – –

• Firstly, water was added to the anode section of the MFC • After the experimental system came to a stable regime
and active sludge / sediment sludge was added to the cath- for the active and sediment sludge, the data obtained by
ode section and made ready. measuring the voltage and current values of the desired
• The copper rod was immersed in the anode part, and the input values were recorded.
copper weave was immersed in the cathode part. • The current density (A/m3 ) was calculated using Eq. (3)
• Three magnetic stirrers were actively operated for the and the power density (W/m3 ) using Eq. (4).
homogeneous distribution of the sludge in the anode
section. ΔT exchanges were also performed with the same
device. 3 Response Surface Methodology (RSM)
• Temperature sensors were immersed in both the sludge
and water. RSM is the most common mathematical modeling and sta-
tistical evaluation method used for the optimization of input

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Table 4 Experimental parameters and measured response values for active sludge

Exp. no R (Ohm) ΔT (°C) V̇ (L/h) A (cm2 ) I (mA) I d (A/m3 ) V (V) P (mW) Pd (W/m3 )

1 2 8 1 100 1.24 0.413 0.002 3.075 1.025

2 2 10 1 100 1.560 0.520 0.003 4.867 1.622
3 2 12 1.5 121 1.150 0.383 0.002 2.645 0.882
4 2 14 1.5 121 1.040 0.383 0.002 2.163 4.033
5 500 8 1 121 0.556 0.185 0.270 154.568 51.523
6 500 10 1 121 0.563 0.188 0.300 158.484 52.828
7 500 12 1.5 100 0.680 0.227 0.340 231.200 77.067
8 500 14 1.5 100 0.610 0.203 0.330 186.050 66.667
9 900 8 1.5 100 0.440 0.147 0.380 174.240 60.000
10 900 10 1.5 100 0.480 0.160 0.400 207.360 73.333
11 900 12 1 121 0.403 0.134 0.388 146.168 48.723
12 900 14 1 121 0.411 0.145 0.430 152.029 64.533
13 10,000 8 1.5 121 0.052 0.017 0.534 27.040 9.013
14 10,000 10 1.5 121 0.056 0.019 0.573 31.360 9.013
15 10,000 12 1 100 0.056 0.019 0.581 31.360 10.453
16 10,000 14 1 100 0.057 0.019 0.593 32.490 10.830

Table 5 Experimental parameters and measured response values for settling sludge

Exp. no R (Ohm) ΔT (°C) V̇ (L/h) A (cm2 ) I (mA) I d (A/m3 ) V (V) P (mW) Pd (W/m3 )

1 2 8 1 100 0.930 0.360 0.001 1.730 0.577

2 2 10 1 100 0.830 0.320 0.002 1.378 0.459
3 2 12 1.5 121 1.067 0.356 0.001 2.277 0.759
4 2 14 1.5 121 0.410 0.002 0.026 1.009 1.009
5 500 8 1 121 0.463 0.154 0.310 107.185 41.543
6 500 10 1 121 0.480 0.160 0.280 115.200 44.367
7 500 12 1.5 100 0.611 0.204 0.305 186.660 62.220
8 500 14 1.5 100 0.640 0.213 0.310 204.800 71.333
9 900 8 1.5 100 0.400 0.133 0.380 144.000 50.000
10 900 10 1.5 100 0.424 0.141 0.390 161.798 56.667
11 900 12 1 121 0.375 0.125 0.372 126.563 44.367
12 900 14 1 121 0.420 0.140 0.380 158.760 68.970
13 10,000 8 1.5 121 0.051 0.017 0.528 26.010 8.670
14 10,000 10 1.5 121 0.052 0.017 0.537 27.040 9.013
15 10,000 12 1 100 0.055 0.018 0.571 30.250 10.083
16 10,000 14 1 100 0.057 0.019 0.588 32.490 10.830

Table 6 Design summary for responses

Response Name Units Obs Mean Min Max Std. Dev Model

I Current (Active Sludge) mA 16 0.5846 0.0520 1.5600 0.4572 Quadratic

Id Current density (Active Sludge) A/m3 16 0.1976 0.0173 0.5200 0.1549 Quadratic
Pd Power density (Active Sludge) W/m3 16 33.8466 0.8817 77.0667 29.9172 Quadratic
V Voltage (Sediment Sludge) V 16 33.8466 0.8817 77.0667 29.9172 Quadratic
P Power (Sediment Sludge) mW 16 82.9468 1.0086 204.8000 74.3814 Quadratic

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Table 7 Equation of regression

models and values of R2 Regression models for active sludge R2 (%)

I  9.62–0.000313R + 0.603T-9.39V̇-0.0969A + 99.49

0.000000R*R-0.00847T*T-0.000002R*T +
0.000285R*V̇-0.000005R*A-0.1180T*V̇-0.00227T*A + 0.0924V̇*A
I d  3.46–0.000016R + 0.1514ΔT-3.15V̇-0.0329A + 99.33
0.000000R*R-0.00213ΔT*ΔT-0.000001R*ΔT +
0.000159R*V̇-0.000003R*A-0.0323ΔT*V̇-0.000490ΔT*A + 0.0298V̇*A
V  -5.999 + 0.000086R-0.0239ΔT + 5.421V̇ + 0.05270A-0.000000R*R-0.000725ΔT*ΔT 99.93
+ 0.000001R*ΔT-0.000030R*V̇ + 0.000001R*A-0.0091ΔT*V̇ +
0.000521ΔT*A-0.04803V̇*A
P  -5570–0.113R + 19.4ΔT + 4704V̇ + 48.7A + 0.000007R*R-1.28ΔT*ΔT + 98.98
0.00020R*ΔT-0.029R*V̇ + 0.00071R*A-5.2ΔT*V̇ + 0.135ΔT*A-41.4V̇*A
P d  -1844–0.0048R-2.9ΔT + 1601V̇ + 16.38A + 98.49
0.000002R*R-0.098ΔT*ΔT-0.000158R*ΔT + 0.0136R*V̇-0.00029R*A-3.00ΔT*V̇ +
0.097ΔT*A-14.20V̇*A

Table 8 Equation of regression models and values of R2 the last stage, surface graphics were created in which the
maximum and minimum points could be determined. In this
Regression models for sediment sludge R2 (%)
way, parameters and levels were decided for the optimum
I  1.0–0.00247R + 0.658ΔT-1.4V̇-0.036A + 91.25 point. At the same stage, ANOVA analysis determined the
0.000000R*R-0.0082ΔT*ΔT + most effective parameters on responses.
0.000007R*ΔT-0.00146R*V̇ + In RSM problems, the mathematical relationship between
0.000034R*A-0.142ΔT*V̇-0.00327ΔT*A + input parameters and the corresponding responses is usually
0.038V̇*A
expressed through the first-order or second-order polynomial
I d  2.72–0.000445R + 0.0104ΔT-1.61V̇-0.0242A + 98.97
0.000000R*R-0.00007ΔT*ΔT +
equations as given in Eqs. (5) and (6) [41–43].
0.000001R*ΔT-0.000218R*V̇ +
0.000005R*A-0.0047ΔT*V̇-0.000077ΔT*A + η  β0 + β1 x1 + β2 x2 + ....... + βn xn (5)
0.0168V̇*A
V  -3.952 + 0.000314R-0.0237ΔT + 3.160V̇ + 99.94
0.03907A-0.000000R*R + 0.001014ΔT*ΔT + 
k 
k 
0.000001R*ΔT + 0.000022R*V̇-0.000001R*A + η  β0 + βi xi + βii xi2 + βi j xi x j + ε (6)
0.0132ΔT*V̇-0.000126ΔT*A-0.02990V̇*A i1 i1 i j
P  -4015–0.122R-1.7ΔT + 3524V̇ + 35.89A + 99.52
0.000003R*R + where, η represents predicted responses (current, current
0.388ΔT*ΔT-0.000478R*ΔT-0.0654R*V̇ + density, voltage, power, power density), β 0 is the constant
0.00168R*A-1.6ΔT*V̇ + 0.013ΔT*A-31.10V̇*A coefficient, and where β i , β ii and β ij represent the coeffi-
P d  -1558–0.0187R-9.0ΔT + 1394V̇ + 14.13A + 98.48 cients of first- and second-order coded input parameters and
0.000002R*R + 0.391ΔT*ΔT-0.000298R*ΔT +
0.0020R*V̇ + 0.00003R*A-2.92ΔT*V̇ +
parameter interactions, respectively.
0.069ΔT*A-12.37V̇*A The method used in the RSM optimization study is shown
schematically in Fig. 5.

3.1 Experimental Design

parameters (independent variables) of any system. In addi-
tion, RSM can easily be used to identify and interpret the In fact, there are many factors that may affect the system,
relationships of input parameter combinations with responses considering the conditions of physical testing. However, a
(dependent variables) [40]. In general, the RSM method is small number of experiments can be performed using the
carried out in three steps (Fig. 4). Latin square array, depending on the reduced factor num-
In the first stage, physical experiments were carried out bers, which represent the effects of all factors and levels.
using the designed experimental parameter combinations for Thus, saving considerable time and costs. The parameters
obtaining response values. Meaning that fewer experiments and levels used in this study are shown in Table 3.
were carried out compared to traditional methods, thus reduc- The response values corresponding to the L 16 (24 × 22 )
ing costs. In the second stage, the model coefficients were orthogonal design of the input parameters are given in
determined by analyzing the obtained experimental data. In Tables 4 and 5.

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0,6
1,6 R2 =99.49% R2 =99.33%

Predicted, Id (A/m3)
Predicted, I (mA)

1,2 0,4

0,8
(a) 0,2 (b)
0,4
0,0
0,0
0,0 0,2 0,4 0,6
0,0 0,4 0,8 1,2 1,6
Observed, I (mA) Observed, Id (A/m3 )

0,6 R =99.94%
2
225
R2 =99.52%
Predicted, V (V)

Predicted, P (mW)
0,4 150

0,2 (c) 75 (d)

0,0 0

0,0 0,2 0,4 0,6 0 75 150 225

Observed, V (V) Observed, P (mW)

80
R2=98.49%
Predicted, Pd (W/m3)

60

40
(e)
20

0 20 40 60 80

Observed, Pd (W/m3)
Fig. 6 Comparison of predicted and real values. a For current parameter (Active sludge). b For current density parameter (Active sludge). c For
voltage parameter (Sediment sludge). d For power parameter (Sediment sludge) e For power parameter (Active sludge)

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Table 9 ANOVA for active

sludge, I Source DF Seq SS Adj MS F P PC%

Model 12 3.1192 0.2599 49.61 0.004 * 99.50

Linear 4 1.7803 0.2071 39.53 0.006 * 56.79
R 1 1.7137 0.7905 150.85 0.001 * 54.67
T 1 0.0097 0.0009 0.17 0.704 0.31
V̇ 1 0.0071 0.0011 0.22 0.674 0.23
A 1 0.0497 0.0021 0.4 0.573 1.59
Square 2 1.1934 0.0128 2.45 0.234 38.07
R*R 1 1.1751 0.0073 1.39 0.324 37.48
T *T 1 0.0184 0.0184 3.5 0.158 0.59
2-Way Interaction 6 0.1455 0.0242 4.63 0.118 4.64
R*T 1 0.0202 0.0009 0.18 0.7 0.64
R*V̇ 1 0.0189 0.0018 0.34 0.601 0.60
R*A 1 0.0010 0.0010 0.2 0.686 0.03
T *V̇ 1 0.0797 0.0139 2.66 0.202 2.54
T *A 1 0.0014 0.0091 1.74 0.279 0.04
V̇ *A 1 0.0243 0.0243 4.64 0.12 0.78
Error 3 0.0157 0.0052
Total 15 3.1349
* Expressions in bold are effective in the 95% confidence interval on the response
df: Degrees of freedom, Seq SS: Sequential sums of squares, Adj MS: Adjusted mean squares F:F-test, P:
probability PC: Percentage contribution

Table 10 ANOVA for active

sludge, I d Source DF Seq SS Adj MS F P PC%

Model 12 0.3573 0.02978 37.28 0.006 * 99.33

Linear 4 0.2015 0.02217 27.75 0.011 * 56.01
R 1 0.1967 0.08444 105.7 0.002 * 54.69
T 1 0.0003 0.00028 0.35 0.596 0.09
V̇ 1 0.0004 0.00044 0.55 0.512 0.12
A 1 0.004 0.00061 0.76 0.447 1.11
Square 2 0.1393 0.00103 1.29 0.394 38.71
R*R 1 0.1381 0.0009 1.13 0.366 38.39
T *T 1 0.0012 0.00116 1.45 0.314 0.32
2-Way Interaction 6 0.0166 0.00276 3.46 0.168 4.61
R*T 1 0.0022 0.00031 0.38 0.579 0.62
R*V̇ 1 0.0016 0.00056 0.69 0.466 0.45
R*A 1 0.0004 0.00043 0.54 0.517 0.12
T *V̇ 1 0.009 0.00105 1.31 0.336 2.49
T *A 1 0.0008 0.00042 0.53 0.519 0.22
V̇ *A 1 0.0025 0.00254 3.18 0.173 0.71
Error 3 0.0024 0.0008
Total 15 0.3597

*Expressions in bold are effective in the 95% confidence interval on the response
See Table 5 for abbreviations

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Table 11 ANOVA for active

sludge, Pd Source DF Seq SS Adj MS F P PC%

Model 12 13,223.2 1101.93 16.33 0.021 * 98.49

Linear 4 2870.8 20.13 0.30 0.863 21.38
R 1 2360.6 32.59 0.48 0.537 17.58
T 1 68.1 17.25 0.26 0.648 0.51
V̇ 1 213.7 5.63 0.08 0.791 1.59
A 1 228.4 6.41 0.09 0.778 1.70
Square 2 8576.4 13.47 0.20 0.829 63.88
R*R 1 8573.9 24.46 0.36 0.59 63.86
T*T 1 2.5 2.48 0.04 0.86 0.02
2-Way Interaction 6 1776 296.01 4.39 0.126 13.23
R*T 1 18.5 6.89 0.10 0.77 0.14
R*V̇ 1 3.7 4.02 0.06 0.823 0.03
R*A 1 3.5 3.50 0.05 0.835 0.03
T*V̇ 1 687 9.01 0.13 0.739 5.12
T*A 1 489.3 16.73 0.25 0.653 3.64
V̇ *A 1 574 574.05 8.51 0.062 * 4.28
Error 3 202.4 67.47
Total 15 13,425.6

*Expressions in bold are effective in the 95% confidence interval on the response
See Table 5 for abbreviations

Table 12 ANOVA for sediment

sludge, V Source DF Seq SS Adj MS F P PC%

Model 12 0.631562 0.05263 425.79 0.000 * 99.94

Linear 4 0.367617 0.031096 251.58 0.000 * 58.17
R 1 0.365696 0.123866 1002.11 0.000 * 57.87
T 1 0.001087 0.000185 1.5 0.309 0.17
V̇ 1 0.000044 0.000015 0.12 0.751 0.01
A 1 0.000791 0.000018 0.15 0.728 0.13
Square 2 0.246385 0.001269 10.27 0.045 * 38.99
R*R 1 0.246122 0.002276 18.41 0.023 * 38.95
T*T 1 0.000263 0.000263 2.13 0.241 0.04
2-Way Interaction 6 0.01756 0.002927 23.68 0.013 * 2.78
R*T 1 0.00067 0.00007 0.57 0.506 0.11
R*V̇ 1 0.000177 0.00001 0.08 0.793 0.03
R*A 1 0.000017 0.000017 0.14 0.733 0.00
T*V̇ 1 0.009501 0.000175 1.41 0.32 1.50
T*A 1 0.004647 0.000028 0.23 0.668 0.74
V̇ *A 1 0.002547 0.002547 20.61 0.02 * 0.40
Error 3 0.000371 0.000124
Total 15 0.631933

*Expressions in bold are effective in the 95% confidence interval on the response
See Table 5 for abbreviations

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Table 13 ANOVA for sediment

sludge, P Source DF Seq SS Adj MS F P PC%

Model 12 82,593.9 6882.82 52.27 0.004 * 99.52

Linear 4 18,084.4 509.02 3.87 0.148 21.79
R 1 11,634.2 276.38 2.1 0.243 14.02
T 1 1947.7 185.34 1.41 0.321 2.35
V̇ 1 2025.9 83.42 0.63 0.484 2.44
A 1 2476.7 64.3 0.49 0.535 2.98
Square 2 51,279.9 34.96 0.27 0.783 61.79
R*R 1 51,241.4 31.42 0.24 0.659 61.74
T *T 1 38.5 38.49 0.29 0.626 0.05
2-Way Interaction 6 13,229.5 2204.92 16.74 0.021 * 15.94
R*T 1 928.7 62.58 0.48 0.54 1.12
R*V̇ 1 220.9 93.37 0.71 0.462 0.27
R*A 1 116.1 116.07 0.88 0.417 0.14
T*V̇ 1 5421 2.56 0.02 0.898 6.53
T*A 1 3787.6 0.29 0 0.966 4.56
V̇ *A 1 2755.3 2755.3 20.92 0.02 * 3.32
Error 3 395.1 131.68
Total 15 82,988.9

*Expressions in bold are effective in the 95% confidence interval on the response
See Table 5 for abbreviations

4 Results and Discussion Table 14 Model precision compares

R-sqr Adj R-sqr MSResidual

Obtained experimental response values were analyzed using
descriptive statistics and the results are given in Table 6. I (Active Sludge) 0.99499 0.97493 0.00524
Table 6 shows the means, minimum–maximum values, and I d (Active Sludge) 0.99334 0.96669 0.0008
standard deviations. The mean values of the responses were
Pd (Active Sludge) 0.98492 0.92462 67.46554
0.5846 mA, current density 0.1976 A/m3 , power density
V (Sediment Sludge) 0.99941 0.99707 0.00012
33.8466 W/m3 , voltage 33.8466 V, and power 82.9468 mW,
P (Sediment Sludge) 0.99524 0.9762 131.6849
respectively, while all models were chosen as quadratic mod-
els instead of linear models due to the likelihood of revealing
higher accuracy in predicting the behavior of two data sets.
When the R2 values of the calculated equations are examined,
it can clearly be seen that the responses are extremely suffi-
4.1 Formulation of Predictive Mathematical Model cient in predicting the reactions closest to reality. When the
R2 values of the experimental response values of the active
Second-order polynomial response prediction functions were and sediment sludge given in Tables 7 and 8 are examined, the
created using the parameters and response values given in output parameters are above 80% and show a strong correla-
Tables 4 and 5. The estimation equation and coefficient of tion between the input parameters and the output parameters.
determination for all responses are given in Tables 7 and 8. It can also be said from the same table that the estimation
The coefficient of determination is the square of the cor- validities of all output parameters are statistically sufficient.
relation between the predicted scores in a data set versus the In Fig. 6, the coherence between the experimental reac-
actual set of scores. It can also be expressed as the square tions and the estimation values produced by the regression
of the correlation between the independent variable and the equations for the prediction of the reactions are shown. It
dependent variable. An R2 value of 70% and above is desir- should be noted here that the most predictive response func-
able. Equation (7) was used to calculate the R2 values. tions for both the active and sediment sludge are shown. The
solid dots in the graphs show the predicted values. The prox-
Explained Variation imity of these dots to the linear lines in the graph shows the
R2  1 − (7) conformity of the estimated values to reality. In Fig. 6, the
Total Variation

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Arabian Journal for Science and Engineering (2022) 47:15705–15725 15717

99 Term
Not Significant 3,18
95 Significant A Factor Name
90 Factor Name CD A R
80 A R BB B ΔT
Percent

70 B ΔT BC C V
60 BD D A
50 C V
40 D A AA
30 D
20 AC
10 C
5 A AD
AB
1 B
-12 -8 -4 0 4
0 2 4 6 8 10 12
Standardized Effect
Standardized Effect
(a) (b)
99 Term 3,18
Effect Type
95 Not Significant A Factor Name
90 Significant CD A R
80 BB B ΔT
Factor Name
BC C V
Percent

70 A R
60 AA D A
50 B ΔT
40 C V D
30 D A AC
20 C
10 AD
5 A BD
AB
1 B
-12 -10 -8 -6 -4 -2 0 2 4
0 2 4 6 8 10
Standardized Effect
Standardized Effect
(c) (d)

99 Term 3,182
Effect Type
95 Not Significant CD Factor Name
90 Significant A A R
B B ΔT
80 Factor Name AD C V
Percent

70 A R
60 AC D A
50 B ΔT C
40 C V D
30 D A
20 AB
BB
10
CD AA
5
BC
1 BD
-5 -4 -3 -2 -1 0 1 2 3 0 1 2 3 4 5
Standardized Effect Standardized Effect
(e) (f)
Fig. 7 a Normal probability chart for current parameter (Active sludge). density parameter (Active sludge). g Normal probability chart for volt-
b Pareto chart for current parameter (Active sludge). c Normal probabil- age parameter (Sediment sludge). h Pareto chart for voltage parameter
ity chart for current density parameter (Active sludge). d Pareto chart for (Sediment sludge). i Normal probability chart for power parameter (Sed-
current density parameter (Active sludge). e Normal probability chart iment sludge). j Pareto chart for power parameter (Sediment sludge)
for power density parameter (Active sludge). f Pareto chart for power

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15718 Arabian Journal for Science and Engineering (2022) 47:15705–15725

99 Term 3,18
Effect Type
95 A Not Significant A Factor Name
90 Significant CD A R
AA B ΔT
80 Factor Name BB C V
Percent

70 A R
60 B D A
50 B ΔT BC
40 C V
30 AB
D A BD
20 AA
10
D
CD AD
5
C
1 AC
-5 0 5 10 15 20 25 30 35 0 5 10 15 20 25 30 35
Standardized Effect Standardized Effect
(g) (h)
99 Term 3,182
Effect Type
95 Not Significant CD Factor Name
90 Significant A A R
AA B ΔT
80 Factor Name B C V
Percent

70 A R
60 BD D A
50 B ΔT
BC
40 C V
30 AB
D A
20 D
10 C
5 AC
AD
1 BB
-3 -2 -1 0 1 2 3
0,0 0,5 1,0 1,5 2,0 2,5 3,0 3,5
Standardized Effect Standardized Effect
(i) (j)
Fig. 7 continued

dots are very close to the linear line, this proving the validity considering the most valid models of active and sedimenta-
of the equations. tion sludge response values. Accordingly, the most effective
parameter on current and current density values concerning
the active sludge was the parameter of resistance at 54.67%
4.2 Analysis of Variance (ANOVA) and 54.69%, respectively (see Tables 9 and 10). In Table 11,
it can be seen that despite having a low impact rate, the most
Quadratic response surface models concerning the most effective parameter on power density was the area interaction
valid response parameters, major and minor effective input with a volumetric flow rate of about 4.28%. While the most
parameters, and interactions were determined using vari- effective parameter in Table 12 was resistance at 57.87%,
ance analysis (ANOVA) in this study. In ANOVA analysis, Table 13 shows that the most effective parameter of the set-
the value of P (significance/probability) indicates whether tling sludge power output parameter was quadratic resistance
the parameter or interaction has an effect on the response. at 61.74%. It can be determined from the ratios that param-
Considering the 95% confidence interval, the parameter is eters other than the specified effective parameters were also
considered to be effective on the response when P < 0.05. somewhat effective for each given ANOVA table. However,
The contribution of each factor on the total variation is indi- since these effects are relatively low, they were not included
cated in percent (PC%) in the last column of the ANOVA in the analyses.
charts. R2 , or the coefficient of determination is introduced, Furthermore, the reasonable agreement between the adj R2
which aids in appraising prediction accuracy, in this case, and pred R2 exemplifies the suitability of the model (Table
through the mathematical equations [44]. 14). The range of the predicted values at the design points
Tables 9, 10, 11, 12, and 13 show the ANOVA table val- were compared with adequate precision to the average pre-
ues calculated for current, current density, power density, diction error with a ratio of  4. The literature shows that
the voltage of sludge, and power responses, respectively, of for these models a ratio greater than 4 is desirable [43, 45].
activated sludge. The analysis given here was carried out by

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Arabian Journal for Science and Engineering (2022) 47:15705–15725 15719

Fig. 8 Effect of test parameters on current response for active sludge. a Surface plot reflecting the impact of the area and flow rate on current.
b Surface plot reflecting the impact of the flow rate and resistance on current. c Surface plot reflecting the impact of the area and resistance on
current

MSResidual was used for their F ratio; however, this may result are significant. The distance that points must be from zero to
in reduced df and less power than anticipated for some tests. be statistically significant depends on the significance level
(denoted by α or alpha). Furthermore, the normal probability
4.3 Normal Probability and Pareto Charts plot of the effects displays negative effects on the left side
and positive effects on the right side of the graph.
The normal probability plot of impacts can be used to deter- Accordingly, it can be seen that in the normal probability
mine the magnitude and direction of the actual impacts graphs given in Fig. 7a,c,e, the most effective parameters
calculated by ANOVA analyses. The normal probability plot and combinations are R for I and I d and V̇ xA for Pd . From
of impacts shows the standardized effects of residual val- the graph displayed in Fig. 7g for the sediment sludge, it
ues based on the distribution of values on a compliance line can be seen that the V output parameter was most highly
for all impacts where it is zero. Standardized effects are t- affected by A, RxR and V̇ xA, respectively. However, as can be
test statistics that test the null hypothesis that the effect is seen from the graphic provided in Fig. 7i, the most effective
zero. Each effect is assigned a point on the chart. The color input parameter e on the P output parameter for the sediment
and shape of the points differ between statistically significant sludge cannot be determined. Parameters assumed to have
and statistically insignificant effects. According to the nor- high effects can be listed as V̇ xA and R.
mal probability plots, the points located closer to the fitted As can be seen from Fig. 7a,c, the R parameter, which has
line are not considered significant, whereas points located an effect on I and I d , for the activated sludge, has a negative
far away from zero on the x-axis tend to be more signifi- effect since it is to the left of zero on the x-axis. To state
cant. All factors and interactions designated by circles are it more clearly, I and I d values decrease as the R parame-
not significant whereas the effects represented by squares ter increases. In parallel, from Fig. 7e, it can be concluded

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15720 Arabian Journal for Science and Engineering (2022) 47:15705–15725

Fig. 9 Effect of test parameters on current density response for active sludge. a Surface plot reflecting the impact of the area and flow rate on current
density. b Surface plot reflecting the impact of the flow rate and resistance on current density. c Surface plot reflecting the impact of the area and
resistance on current density

that the combination of parameters RxR and V̇ xA, which Accordingly, it is understood that all parameters and inter-
are effective on Pd for active sludge and are effective on V actions for the activated sludge in Fig. 7b, d and in Fig. 7h for
for sediment sludge, have a negative standard effect. How- the sediment sludge that cross the reference line are statis-
ever, the R parameter which on the right side of the x-axis in tically significant. Whereas the most effective parameter or
the graph given in Fig. 7g has a positive effect on response, combination on V cannot be understood from the sediment
indicating that increasing the R value increases the V output sludge normal probability graph, the Pareto chart in Fig. 7j
value. shows that this is V̇ xA as it is closest to the dashed line while
The Pareto chart shows the absolute values of the standard- not exceeding the reference line at 3.182.
ized effects from the largest effect to the smallest effect. The
significance of the factors and their interactions was obtained
by employing a Student’s t-test with a 95 percent confidence 4.4 Graphical Optimization
interval. The chart also plots a reference line to indicate
which effects are statistically significant. Values exceeding Graphic optimization results allow for visual inspection to
the reference line are considered significant. Because the select optimum values for the effective parameters. With the
Pareto chart displays the absolute value of the effects, large help of contour and 3D surface graphics created depend-
effects can be determined; however, which effects increase ing on the regression equations obtained, the effects of the
or decrease the response cannot be defined. experimental parameters on the reactions and the optimum
parameter combinations can be clearly understood. The opti-
mum point can be found visually at the intersection of the

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Arabian Journal for Science and Engineering (2022) 47:15705–15725 15721

Fig. 10 Effect of test parameters on power density response for active sludge. a Surface plot reflecting the impact of the area and flow rate on power
density. b Surface plot reflecting the impact of the flow rate and resistance on power density. c Surface plot reflecting the impact of the area and
resistance on power density

input parameter values that maximize the response parameter and parameter A should be kept at the lowest level for the
values. Contour and surface plots were created by consider- maximum value of the I parameter.
ing the regression models of the activated sludge (I, I d and Graphs showing the relation of the R input parameter (see
Pd ) and sediment sludge (V and P) output parameters in Fig. 7), which is known to be effective on the I d reaction
which these parameters have the highest estimation valid- parameter of the active sludge, with the other effective param-
ity. Exclusive of the input parameters used for each of the eters (V̇ xA) are shown in Fig. 9. According to this, it can be
graphs, parameters were accepted as fixed values by Sta- understood from Fig. 9b and c that the relationship between
tistica 10 statistical software. The contour and 3D graphics R-A and R-V̇ with the highest I d value change interval is R,
created by using the first two input parameters most effective which is the main factor affecting the I d value. It is possi-
on the output parameter found by ANOVA analysis are given ble to see the relationship between the V̇ and A parameters,
in Fig. 8. which have a limited effect, in Fig. 9a. Consequently, in order
When the graphics created after these assumptions are to reach the desired high I d value, the levels of the R and A
examined, the maximum value of I in Fig. 8b,c of the active parameters should be minimum with a maximum V̇ param-
sludge is possible with the lowest value of the R parameter. eter level. This situation coincides with the inference made
It may be misleading to use these graphs regarding the effec- for I.
tiveness of the given parameters associated with R. Inspection From the graphs for active sludge provided in Fig. 10, the
of Fig. 8a, which gives the relationship between V̇ and A, condition required to achieve the highest Pd parameter value
indicates that parameter V should be kept at the highest level is a maximum of R and V̇ parameters with a minimum of the
A parameter.

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15722 Arabian Journal for Science and Engineering (2022) 47:15705–15725

Fig. 11 Effect of test parameters on voltage response for sediment sludge. a Surface plot reflecting the impact of the area and flow rate on voltage.
b Surface plot reflecting the impact of the flow rate and resistance on voltage. c Surface plot reflecting the impact of the area and resistance on
voltage

From the graphics displayed in Fig. 11b,c for sediment 5 Conclusions

sludge, it can be easily seen that the most effective R input
parameter to maximize the V parameter is at 6000 Ohm lev- In this research, the performance of active and sediment
els. Figure 11a shows that V̇ and A values should be kept at sludge in electricity production in an MFC was predicted by
maximum levels to reach the highest V value. the optimization of operating conditions. Using response sur-
The sediment sludge graphs in Fig. 12 indicate that for face methodology, involving Taguchi orthogonal design and
maximum P, the parameters of R and A should be at the regression of analysis, quadratic mathematical models were
highest level and the V̇ parameter should be at the lowest derived for the prediction of current, current density, voltage,
level. power, and power density. The results suggest that the mod-
In addition, the entire surface charts with gutter struc- els were in sufficient agreement with the experimental data.
ture given above show that the input parameters quadratically Surface charts were created by considering the models of the
affect the response parameters. active sludge (I, I d and Pd ) and sediment sludge (V and P)
output parameters having the highest estimation validity. The
3D plots clearly demonstrate that the most effective param-
eter on all responses at optimized conditions was resistance.
Therefore, the implication is that the method applied in this

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Arabian Journal for Science and Engineering (2022) 47:15705–15725 15723

Fig. 12 Effect of test parameters on power response for sediment sludge. a Surface plot reflecting the impact of the area and flow rate on power.
b Surface plot reflecting the impact of the flow rate and resistance on power. c Surface plot reflecting the impact of the area and resistance on power

study can be safely used for similar studies by comparing 3. Zinadini, S.; Zinatizadeh, A.A.; Rahimi, M.; Vatanpour, V.;
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by Selcuk University Scientific Research Projects with the project num- 4. Mardanpour, M.M.; Esfahany, M.N.; Behzad, T.; Sedaqatvand, R.:
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