Title


Indonesian Journal of Environmental
Management and Sustainability
e-ISSN:2598-6279 p-ISSN:2598-6260

Research Paper

Tofu Wastewater (TWW) Treatment and Hydrogen (H2) Production by Using A

Microbial Electrolysis Cell (MEC) System

Ibdal Satar1*, Muhammad Mar’ie Sirajuddin1, Adi Permadi2, Sukma Latifatunnajib1

1Department of Food Technology, Faculty of Industrial Technology, Universitas Ahmad Dahlan, Bantul, Yogyakarta, 55191, Indonesia
2Department of Chemical Engineering, Faculty of Industrial Technology, Universitas Ahmad Dahlan, Bantul, Yogyakarta, 55191, Indonesia

*Corresponding author e-mail: ibdal@tp.uad.ac.id

Abstract
High organic pollutant in tofu wastewater (TWW) raises a negative impact on environmental sustainability and health.
Therefore, the TWW must be treated before it is discharged into the environment. Microbial electrolysis cell (MEC) is one of
the green technologies that can be used to treat wastewater and generate hydrogen as well. This work tries to investigate
the performance of MEC based on the decrement of organic pollutants in TWW. Some important parameters of organic
pollutants in TWW such as chemical oxygen demand (COD), biological oxygen demand (BOD), total suspended solids (TSS),
total dissolved solids (TDS), total solid (TS), and pH were evaluated before and after MEC operation. The results showed
that the COD and BOD levels decreased around 56% and 35% while pH increased from 7.90 to 7.16. Additionally, the TSS,
TDS, and TS decreased by around 35.0%, 45.5%, and 33.2%. In addition, the optimum hydrogen yield (YH2) and hydrogen
production rate (QH2) were obtained at 114 ± 0.1 mL H2/g COD 360 ± 20 mL H2/L/d. Overall, the MEC system could be
used to reduce the level of organic pollutants in TWW and generated H2 at the same time.

Keywords
TWW, Organic Pollutants, pH, COD, MEC

Received: 14 November 2022, Accepted: 21 February 2023

https://doi.org/10.26554/ijems.2023.7.1.13-19

1. INTRODUCTION

Nowadays, environmental sustainability become the main
problem that has to solve immediately. There are two
environmental issues namely excess of pollutant gases (i.e.,
CO2, CO, SOx, and NOx) in the atmosphere and high
amount of wastewater in the environment (Manisalidis et al.,
2020; Igwe et al., 2017). Excess gases and high amounts of
wastewater are caused by human activities, population and
economic growth (Lin et al., 2022). It is well known that the
use of fossil fuels in industry activities and vehicles increases
air pollution over time (Lelieveld et al., 2019). In Indonesia
(represented by Jakarta city), according to the air quality life
index (AQLI) data that the quality of air in Indonesia since
2016 is getting worst (Greenstone and Fan, 2019; Istiqomah
and Marleni, 2020). In fact, air quality index (AQI) in
Indonesia is ranked 17th worst from 118 countries in the
world for the period of 2021. Whereas, approximately 70%
of wastewater is generated by households and the rest is
produced by other sectors (industries, public facilities, office
buildings, etc) (Igwe et al., 2017; Wulan et al., 2022).

As presented above, pollutant gasses and wastewater are
two main issues currently focused on by researchers. The

number of pollutant gasses can be eliminated by replac-
ing fossil fuel with alternative fuel (i.e., H2) (Cloete et al.,
2022). In addition, the wastewater issue can be solved by
performing chemical, physical, and/or biological approaches
(Ye and Li, 2023; Gautam et al., 2022). Based on these
environmental contexts, the authors suggest choosing the
biological method to solve both gasses and wastewater is-
sues. There are two biological methods such as fermentation
and microbial electrosynthesis technologies (METs) which
generally can be applied to treat wastewater and to generate
biogas and/or biohydrogen and electricity (Son et al., 2021).
Among these biological approaches, the METs are more
interesting compared to the fermentation due to the METs
have good efficiency and generate biogas.

One of the METs technologies is microbial electrolysis
cells (MEC). Principally, MEC consists of two parts which
are the anode and cathode side. The anode side converts
the organic substrate to volatile fatty acids (VFAs) and
electrons (e−) (see Equation 1), while the cathode side
generates hydrogen (H2) from protons (H

+) and electrons
(see Equation 2). Due to the hydrogen production being
unspontaneous reactions so the additional energy must be

https://doi.org/10.26554/ijems.2023.7.1.13-19


Satar et. al. Indonesian Journal of Environmental Management and Sustainability, 7 (2023) 13-19

supplied into the MEC system. Generally, the amount of
additional energy in the MEC systems is required in the
range of 0.20 - 0.80 V (Call and Logan, 2008).
Anode: Organic substrate CH3COO

− + 4H2O → 2HCO3−
+ 9H+

Cathode: 8H+ + 8e− → 4H2
Overall: CH3COO

− + 4H2O → 2HCO3− + H+ + 4H2
Theoretically, the performance of MEC depends on some

parameters such as anode and cathode performance, type
and characteristics of substrates, and amount of additional
energy input into the system (Apostolopoulos et al., 2021).
The substrate with a high simple organic acid, rich nutrient,
and neutral pH are very appropriate for generating H2. In
general, the additional energy at the optimum condition is
obtained at approximately 0.80 V (Meshram and Jadhav,
2017). This work tries to investigate the performance of f
MEC system based on the TWW quality and hydrogen pro-
duction when using TWW as substrate. Some parameters
such as the change of pH, COD, BOD, TSS, H2 yield, and
H2 production rate are evaluated.

2. EXPERIMENTAL SECTION

2.1 Materials
Materials such as acrylic blocks and cation exchange mem-
brane (CEM, CMI 7000s) were obtained from Fuel Cell In-
stitute Laboratory (FCIL) University Kebangsaan Malaysia
(UKM). Whereas, all chemicals were purchased from online
shops. Some chemicals needed in this work were hydrochloric
acid (HCl, analytical grade), sodium acetate (CH3COO-Na,
99%), potassium chloride (KCl, analytical grade), ammo-
nium chloride (NH4Cl, analytical grade), potassium dihydro-
gen phosphate (KH2PO4, analytical grade) and di-potassium
hydrogen phosphate (K2HPO4, analytical grade), ethanol
(C2H5OH, 99.5%) and deionized water (DW).

The spectrophotometer (Thermo-scientific) was used to
determine the chemical oxygen demand (COD). The pH
tester (H198103, Hanna Instruments) was used to measure
the initial and final pH of TWW. The power supply (YIHUA
1502 DD, DC power supply) was used to supply the addi-
tional voltage to the MEC system. The scanning electron
microscope and electron dispersive X-ray (SEM-EDX, JEOL
JSM 5800) was used to characterize the electrode surfaces
before and after the MEC system run.

2.2 Methods
2.2.1 Tofu Wastewater (TWW) Preparation and

Identification
A total of 10 liters of TWW sample was collected from the
tofu home industry nearby Imogiri Barat. Some physico-
chemical parameters such as pH, chemical oxygen demand
(COD), biological oxygen demand (BOD) and total sus-
pended solid (TSS), total dissolved solids (TDS), and total
solid (TS) were measured by using APHA, Winkler, and
Spectroscopy methods, respectively. The characterizations
of fresh TWW were presented in Table 1. Initially, the

Table 1. Characteristic of TWW Sample

Parameters Average Values Unit

pH 7.90 -
TDS 4546.67 mg/L
TS 7920.00 mg/L
TSS 1903.33 mg/L
BOD 1400.10 mg/L
COD 1066.67 mg/L

TWW sample was filtered by using a filter cloth to sepa-
rate the solution and active sludge before it was used in
the MEC system. Then, a filtered solution of TWW was
kept and used as a substrate for further work. Meanwhile,
the active sludge of TWW was used as a mixed-culture of
electroactive bacteria (EAB) source in the anode before the
MEC system was operated. It was well known that the EAB
must previously be attached and enriched at the anode to
enhance the system performance (Satar et al., 2021).

2.2.2 Preparation of Electrodes
The preparation of the electrode was addressed to remove the
physical impurities on the material surface. The electrode
preparation was performed in acid and alkali solutions as
described by Satar et al. (2021). The Electrodes (both anode
and cathode) were fabricated from the graphite foam (GF)
materials with a configuration of 2.5 cm × 2.5 cm (Figure
1). Before it was used in the MEC system, the electrodes
were immersed in the 250 mL HCl (1 M) for 1 hour and
washed using deionized water (DW) three times. Then, the
electrodes were immersed in 250 mL KOH (1 M) for 1 hour
and washed using DW three times. Lastly, the electrode
materials were dried in a vacuum oven at 80 oC overnight
(Satar et al., 2018).

Figure 1. Graphite Foam Electrodes; Anode (Left) and
Cathode (Right) Materials

© 2023 The Authors. Page 14 of 19



Satar et. al. Indonesian Journal of Environmental Management and Sustainability, 7 (2023) 13-19

2.2.3 Dual Chamber MEC Set-up and Operation
The MEC system was designed in the dual chamber types
which were fabricated from acrylic block materials. The
MEC system was divided into the anode and cathode cham-
bers with an active volume of 50 mL, respectively (Satar
et al., 2020a). Both anode and cathode chambers were sep-
arated by CEM (CMI, 7000s) to prevent a short circuit in
the system. The anode and cathode materials were installed
with the iron sheet to facilitate the current transfer from a
power supply to the system. The schematic and photograph
of the MEC system were presented in Figure 2.

Figure 2. The Design of the MEC System; (a) Schematic
and (b) Photograph of Dual Chamber MEC System

The MEC system was operated at room temperature
and fully maintained in anaerobic condition to avoid contam-
ination on EAB at the anode and hydrogen gas (H2) release
from the cathode (Bora et al., 2022). Before the MEC was
operated, the anode was initially enriched with EAB using
active sludge of TWW while the cathode was without chem-
ical catalysts. During the enrichment process, the 1.0 g/L
CH3COONa solution was used as substrate at the anode
while 1 M KCl as anolyte at the cathode. A direct current
power supply (YIHUA 1502 DD) was needed to supply 1.0 V
of additional energy into the system because hydrogen pro-
duction is an unspontaneous reaction. An external resistor
(R = 1.0 ohm) was installed at the MEC system to convert
a voltage (V) from the power supply to current (I). Once
the H2 gas was produced from the system, the CH3COONa
substrate in the anode was replaced with TWW solution.
The operation of the MEC system was stopped when the
H2 was stably produced in three consecutive runs.

2.2.4 Gas Analysis and Calculation
The generated H2 gas was collected using a 25 mL syringe
for three consecutive cycles. The amount of generated H2
was calculated based on the purity of H2 production by using
dual chamber MEC (69–77%). The nH2 is a mole of H2 that
is calculated based on the ideal gas formula at standard
conditions (Equation 1). The MH2 is the molecular mass
of H2 (2 g/mol). The COD removal (∆COD) is calculated
based on the total COD value at the beginning (COD0)
minus the COD value at the final (CODt). The H2 yield
(YH2) and H2 production rate (QH2) were calculated by

Table 2. Summary of TWW Characterization at the Final
of MEC Run

Parameters Average Values
Standard (Peraturan
Daerah DIY, 2016)

pH* 7.16 ± 0.1 6.0 – 9.0
TDS 2476.67 mg/L -
TS 5293.33 mg/L -
TSS 1236.67 mg/L 200 mg/L
BOD 490.04 mg/L 150 mg/L
COD 469.33 mg/L 300 mg/L

Note: * = the pH of TWW after the MEC system run

using Equation 2 and Equation 3, respectively (Satar et al.,
2020b).

nH2 =
PV

RT
=

VH2
22.4L

(1)

YH2 =
nH2 × MH2
Vl × ∆COD

× 100% (2)

QH2 =
VH2

VA × t
(3)

Figure 3. Images of Surface Morphologies of Electrode and
Element Compositions: (a) Electrode Image, (b) EDX
Spectra Data Before the MEC System Run, (c) Anode and
(e) Cathode Morphologies, (d) EDX Spectra of the Anode
and (f) EDX Spectra of Cathode After the MEC System
Run

© 2023 The Authors. Page 15 of 19



Satar et. al. Indonesian Journal of Environmental Management and Sustainability, 7 (2023) 13-19

3. RESULTS AND DISCUSSION

3.1 Electrode Images Before and After the Experi-
mental Run

The surface morphologies and element composition of elec-
trodes before and after the MEC system run were presented
in Figure 3. Either anode or cathode surfaces before the
MEC system run was observed clean as shown in the typical
image of GF surface (Figure 3(a)). Based on the SEM-EDX
results, the main element on the electrode surface was car-
bon (C) with 100% of relative composition (Figure 3(b)).
Meanwhile, the morphologies of anode and cathode surfaces
after the experiment run were presented in Figure 3(b) and
(d) respectively. The C compositions at the anode and cath-
ode surfaces were observed to be reduced to 70.1% (Figure
3(d)) and 66.4% (Figure 3(f)), respectively. These facts
might due to the presence of other chemical components
or impurities at the anode and cathode surfaces as a result
of the oxidation and reduction reactions at their surfaces.
Furthermore, the MEC systems were continuously supplied
with the additional voltage from a power supply, thus the
oxidation and reduction reactions occur during the MEC
system operation.

According to some references, the presence of impuri-
ties on the electrode surface inhibits the bio-electrochemical
reaction due to the current flow being slow (Apostolopou-
los et al., 2021; Satar et al., 2020b; Carmona-Mart́ınez
et al., 2015). Consequently, the MEC performance grad-
ually dropped after three months of the system operation
(Carmona-Mart́ınez et al., 2015). Therefore, the anode and
or cathode of the MEC system is generally replaced with a
new one before it is used again.

3.2 TWW Characterization
The quality of TWW before and after being treated with
the MEC system was evaluated to ensure whether the char-
acteristics of TWW meet the national standard of Indonesia
or not. The national standard of Indonesia concerning the
TWW quality regulated by Minister of Environment of the
Republic of Indonesia No. 5, 2014 was referred to in this
work. Some parameters of fresh TWW such as pH, COD,
BOD, TSS, TDS, and TS were detailly measured (Table
1). Based on Table 1 the pH of fresh TWW was observed
around 7.90. According to the Regulation of Government
of Yogyakarta Special Region, No. 7, 2016, concerning the
wastewater standard (Peraturan Daerah DIY, 2016), the
pH of wastewater must be around 6.0 – 9.0 before discharge
to the environment. Based on this pH condition indicates
the fresh TWW can be discharged into the environment. It
is well known the low pH of wastewater cause a negative
impact on the pH of the soil, consequently, the nutrition
absorption by plants is inhibited (Rachmawati and Wardiy-
ati, 2018). Furthermore, the low-pH wastewater may also
interfere the marine life and aquatic biota (Xu et al., 2023).
Hence, the fresh TWW must be primarily treated before it
discharges to the environment. In this work, the pH of fresh

TWW was treated by using the MEC system. The final pH
of TWW was quite decreased to 7.16. This fact indicates
the TWW still be safely discharged into the environment.
Overall, the characterization of TWW at the final MEC run
was presented in Table 2.

In addition to pH, the TSS, TDS, and TS were evaluated
to ensure the physicochemical properties of TWW meet
with the Regulation of Government of Yogyakarta Special
Region No 7 2016. The physical properties of TSS, TDS, and
TS involve the presence of solid impurities in TWW. The
high number of solid impurities in TWW inhibits sunlight
and oxygen entry into the water, consequently, marine life
and or aquatic biota are disrupted. In this work, the TSS,
TDS, and TS of the treated TWW decreased around 35.0%,
45.5%, and 33.2% respectively. Due to the TSS, TDS and
TS were higher than the TS standard (200 mg/L), so further
treatment must be conducted. For example, the filtration
process can be used to reduce these solid impurities.

Furthermore, the COD value is principally indicated
the number of organic matters that exist in a sample. In
addition, the high BOD value indicates a low number of
dissolved oxygen (DO) exists in the sample. Based on two
conditions which are the high COD and BOD values harm
the environment. This work shows the BOD and COD values
of TWW were changed after it was treated with the MEC
system. The BOD was decreased from 1400.10 mg/L to
490.0 mg/L (decrement of 35.0%), while the COD value was
decreased from 1066.67 mg/L to 469.3 mg/L (decrement
of 56.0%). The presence of EAB at the anode of MEC
consumes the organic matter of TWW during the MEC
operation. However, the final BOD and COD values were
quite high than the regulation, hence, the TWW must be re-
treated more either chemical, biological or physical method.
The high COD and BOD values might be due to the low
performance of the anode. The performance anode is very
associated with the ability of EAB to consume the organic
pollutants. In this work, the MEC system was only operated
under normal conditions, consequently, the EABs might be
poisoned by oxygen so their abilities were low. Therefore,
the MEC system must fully be maintained at anaerobic
conditions.

3.3 Biohydrogen Production and MEC Performance
The phenomenon of the current trend at the enrichment and
steady-state stages of the MEC system run was shown in
Figure 4. At the enrichment stage in the MEC anode, the
generated current was observed very low (0.02 - 0.03 mA)
for more than 20 days. There were very low gases produced
from the system at the enrichment stage. Whereas, the
current production at the steady state was observed higher
(0.4 ± 0.1 mA) for 10 days. However, the current generation
was low due to there were no catalysts used at the cathode
of MEC, so the YH2 and QH2 were quite low. Therefore, the
chemical and or biological catalysts must be applied onto the
cathode surface to enhance the H2 evolution reaction (HER).

© 2023 The Authors. Page 16 of 19



Satar et. al. Indonesian Journal of Environmental Management and Sustainability, 7 (2023) 13-19

Table 3. Summary of H2 Production from Dual Chamber MEC Using TWW Substrate and Selected References

Parameters
MEC Design

Dual Chamber Dual Chamber Dual Chamber Dual Chamber

Anode-cathode GF-GF GF-GF/Pt GF-CC/SSW GF-Ni 201
Substrate TWW EGF EGF Acetate
Eap (V) 1.0 1.0 1.0 0.9

YH2 (mL H2/gCOD) 114.1 ± 0.1 553.6 ± 53.5 330 ± 30 -
QH2 (mL/L/d) 360 ± 20 590 ± 10 520 380 ± 40
References This work (Satar et al., 2020b) (Ribot-Llobet et al., 2013) (Selembo et al., 2009a)

Note: CC/SSW = carbon cloth combined with stainless wool; GF = graphite foam; GF/Pt = graphite coated with Pt
catalyst; Ni 201 = nickel allow 201; EGF = effluent of glucose fermentation.

Principally, the overpotential at the cathode is high, hence,
the catalysts are required to increase the total amount of
H2 production (Park et al., 2022).

Figure 4. Typical Current Generation During H2
Generation for 36 Days of the Experiment Run

Hydrogen (H2) produced from the MEC system was
measured by using a syringe (50 mL) without further GC
analysis. Based on some literature the purity of H2 produced
from the dual-chamber MEC generally reached around 69-
77% (Satar et al., 2020b; Chae et al., 2010; Selembo et al.,
2009b; Kadier et al., 2018). Based on these references,
hence, the purity of H2 produced in this work can also
be assumed around 70%. After three consecutive cycles
of biogas production, the total amount of gas volume was
obtained at 68.1 ± 2.0 mL (or 48.1 mL of assumed H2) for
24 hours. The COD removal (∆COD) along with the system
operation was obtained at 597 ± 10 mg/L. According to
Equation 1, the YH2 was calculated at around 8.87% (or
114.1 mL H2/g COD). Meanwhile, the QH2 was calculated
at around 360 ± 20 mL/L/d. This work demonstrated that
the H2 can be generated by a dual chamber MEC system

using TWW as substrate. However, the performance of
MEC (without cathode catalyst) using TWW as substrate
was quite lower than that of the selected references in Table
3. As mentioned above this issue might be due to the rate
of H+ reduction reaction to H2 at the cathode being quite
slow.

4. CONCLUSIONS

The dual chamber MEC system was successfully applied to
treat TWW and generated H2 at the same time. The TSS,
TDS, TS, COD, and BOD values of the final TWW were
reduced by approximately 45.5%, 35.0%, 33.2%, 56%, and
35%, respectively, lower than that of fresh TWW. Meanwhile,
the pH decreased from 7.90 to 7.16. In addition, the YH2
and QH2 were obtained at approximately 8.87% (99.5 ± 0.1
mL H2/g COD) and 360 ± 20 mL/L/d, respectively. Due to
there being no catalyst on the cathode so the performance
of dual chamber MEC was lower than that of the selected
reference. In the next research, the cathode of the dual
chamber MEC system will be coated with biological and or
chemical catalysts to improve the YH2 and QH2.

5. ACKNOWLEDGEMENT

The authors gratefully acknowledge the financial support
given by the Universitas Ahmad Dahlan (UAD) via research
sponsorship of PDP-341/SP3/LPPM-UAD/VI/2022.

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	INTRODUCTION
	EXPERIMENTAL SECTION
	Materials
	Methods
	Tofu Wastewater (TWW) Preparation and Identification
	Preparation of Electrodes
	Dual Chamber MEC Set-up and Operation
	Gas Analysis and Calculation


	RESULTS AND DISCUSSION
	Electrode Images Before and After the Experimental Run
	TWW Characterization
	Biohydrogen Production and MEC Performance

	CONCLUSIONS
	ACKNOWLEDGEMENT