Journal of Environmental Chemical Engineering 6 (2018) 110–118

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Journal of Environmental Chemical Engineering

journal homepage: www.elsevier.com/locate/jece

In-situ prevention of hydrogen sulfide formation during anaerobic digestion T

using zinc oxide nanowires
⁎
Robert Lupitskyy, Dania Alvarez-Fonseca, Zachary D. Herde, Jagannadh Satyavolu
Conn Center for Renewable Energy Research, University of Louisville Louisville, KY 40208, United States

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

Keywords: The utilization of Anaerobic Digestion (AD) technology for biogas and energy generation is often hindered by
Hydrogen sulfide high concentration of sulfur-containing compounds in wastewater and consequential hydrogen sulfide (H2S) in
Biogas biogas. In this work, we demonstrated a process, where independent mechanisms of anaerobic microorganisms
Anaerobic digestion and an inorganic reactive adsorbent (zinc oxide nanowires) worked synergistically to remove soluble sulfide
Metal oxide nanowire
(HS−) and prevent H2S formation during AD. Using a model aqueous sodium sulfide (Na2S) system, the nano-
Zinc oxide
Reactive adsorbent
wires (dosage 1 g/L) effectively removed sulfides from the Na2S solution to a maximum of 625 mg S2−/L per
gram of ZnO. During 24 h and 3-day long (involving three consecutive sulfate feeding cycles) AD studies using
anaerobic microbial medium (2 g/L SO42− and 1 g/L ZnO), no decrease in methanogenic activity and biogas
production were observed using the ZnO nanowires, indicating that the nanowires can reduce the sulfide toxicity
during AD. The post-process analysis of the recovered nanomaterial using energy-dispersive X-ray spectroscopy
and X-ray diffraction showed the presence of sulfur and zinc sulfide (ZnS), respectively, validating HS− removal
by the nanowires. This process intensification approach of combining AD and H2S removal into a single process
step will help extend AD technology to high sulfate containing wastewaters.

1. Introduction preferably, minimizing the formation of H2S in biogas [6–8] is required

Anaerobic digestion (AD) technology is widely used to treat muni-
cipal waste, as well as industrial and food processing wastewaters prior
to discharge. Biogas, a major product of AD, is the most sustainable of
biofuels and is in a starting phase of an exponential market growth
curve. It can be used to produce electricity, heat, and transport fuel [1].
Also, the semi-solid byproduct of this process (digestate) has a high
content of nutrients, which can be used in agriculture directly as a
fertilizer or processed into compost to increase its quality [2,3]. Thus,
anaerobic digesters offer many potential benefits to both the industry
and the environment. Therefore, the demand on the AD technology to
be robust across a wide range of incoming waste streams and the need
for biogas of consistent quality to feed gas generator sets for electricity
production are increasing.
The biggest obstacle for successful application of AD for energy
generation is the presence of hydrogen sulfide (H2S) in biogas. Elevated
levels of H2S in biogas are not only corrosive to the piping, turbines,
and other equipment associated with the biogas industry [4], but also
can lead to harmful sulfur dioxide emissions during combustion of the
biogas. Concentrations of H2S in raw biogas are typically between
50 mg/L and 5000 mg/L. Removing H2S from biogas [5] or, more
to protect downstream equipment such as turbines and fuel cells. In
terms of equipment tolerances, H2S concentrations must be between
200–500 mg/L for combustion of biogas in an internal combustion
engine (co‐generation). Direct injection of biogas into the natural gas
pipeline network as “upgraded bio-methane” (natural gas equivalent)
would require H2S reduction down to below 4 mg/L.
Hydrogen sulfide in biogas is produced by sulfate-reducing bacteria,
commonly present in the anaerobic sludge. Sulfate reducing bacteria
grow in the presence of sulfate, sulfite, or thiosulfate in the incoming
waste stream, producing soluble sulfide (Table S1 in Supplementary
materials). The soluble sulfide, under proper conditions, is liberated as
H2S gas (Eq. (1)). One possible option to reduce the H2S formation is to
remove soluble sulfide (HS−) as it is being formed.
HS−
aq + Haq → H2Sg
+
(1)
Some waste streams contain high concentration of sulfates, which
will lead to elevated levels of H2S in biogas as well as a reduction in the
efficiency or an irreversible inhibition of the anaerobic digestion
[7–12]. In order for such waste streams to be treatable, it is crucial to
develop techniques that minimize the formation of soluble sulfide in
situ.

⁎
Corresponding author.
E-mail address: jagannadh.satyavolu@louisville.edu (J. Satyavolu).

https://doi.org/10.1016/j.jece.2017.11.048
Received 17 August 2017; Received in revised form 30 October 2017; Accepted 16 November 2017
Available online 22 November 2017
2213-3437/ © 2017 Elsevier Ltd. All rights reserved.
R. Lupitskyy et al.
Table 1
Methods of in situ reduction of sulfides during AD.
# Method
1 pH elevation to reduce formation of toxic undissociated H2S
2 Precipitation of sulfides using iron salts
3 Oxidation of sulfides using chemical oxidation agents
4 Chemical inhibition of sulfate-reducing bacteria
5 Biological sulfide removal using sulfide-oxidizing bacteria
(micro-aeration)
6 Biological sulfide removal using sulfide-oxidizing photosynthetic
bacteria
7 Biological denitrifying sulfur removal
2. H2S removal strategies
There are two general H2S removal strategies from biogas: 1) post-
treatment of biogas after anaerobic digestion and 2) in-situ removal
during anaerobic digestion. The most common commercial methods for
hydrogen sulfide removal from the biogas are: biological filters, iron
oxide pellets, impregnated activated carbon, and scrubbing using
water, sodium hydroxide, etc. Scrubbing and recirculating digester
biogas is a common practice to reduce the content of H2S in biogas [5].
However, this is an expensive option requiring additional process
equipment at a significant cost. Various methods have been used for in
situ reduction of sulfides during the digestion process, as listed in
Table 1.
Chemical methods (Table 1, methods 1–4), although efficient in
mitigating the negative effect of sulfides, require constant introduction
of chemicals, as well as additional equipment and process steps. Thus,
these methods are unsustainable and add significantly to the cost of the
AD treatment [17]. Recently, several biological methods of sulfide re-
moval have emerged as a promising alternative (Table 1, methods 5–7;
detailed discussion of the methods is available in Supplementary ma-
terials). However, these methods are in their early stages of develop-
ment and require a significant research effort to become a robust
technology. Thus, there is still a great need for a cost-efficient, sus-
tainable, and robust technology for sulfide removal during AD.
In this work, a hybrid process was developed for in-situ H2S removal
during AD by adding a nanostructured reactive adsorbent consisting of
zinc oxide nanowires (ZnO NWs). ZnO has been used as an efficient and
regenerable adsorbent in the high temperature desulfurization of coal-
derived fuel gas [18] and syngas [19]. ZnO is favorable among the other
metal oxides (e.g., iron oxide) due to its high equilibrium constant for
the ZnO-H2S reaction as well as its high reactivity and the ability of the
resulting ZnS to be regenerated. The advantages of using ZnO NWs
instead of ZnO powders are due to their monocrystalline nature and
faceting, which improves the reactivity of the material. They are also
resistant to sintering and capable of maintaining a high surface area.
The reactive adsorbent in the form of pellets is placed in a water-
permeable pouch (mesh 200, nylon) for easy recovery and introduced
into the AD medium during the anaerobic digestion process. While the
sulfate reducing bacteria are converting the sulfates in wastewater to
HS−, the nanowires are adsorbing the generated HS−, preventing the
formation of hydrogen sulfide gas. According to the following equa-
tions, the adsorbed HS− reacts with ZnO to form ZnS, and water is
formed as a byproduct.
HS− + ZnO → ZnS + OH‐
− using a commercial hydrogen sulfide test kit, Model HS-WR (Hach
OH + H → H2O
+
Our approach is expected to minimize the formation of H2S in
biogas by adsorbing HS− in the waste water under ambient conditions,
increase the biogas yield, reduce toxicity to the biomass, and make the
AD operation more stable over a wide range of sulfate levels. This
process can be a retrofit to the existing AD systems and does not add
significantly to the operating cost.
Journal of Environmental Chemical Engineering 6 (2018) 110–118
Previous studies by Petzold et al. [20] describe the efficiency of a
nanostructured catalyst in the hydrodesulphurization of diesel. In their
work, it was shown that at elevated temperature, Ni nanoparticles
Reference
decorating ZnO nanowires catalyze the cleaving of cyclic sulfur com-
[10] pounds and sequester the released sulfur in the form of NiS. The ZnO
[11] nanowires, in the presence of hydrogen, react with sulfur atoms from
[11,12]
NiS sites forming ZnS and regenerating the nickel catalyst. While this
[13]
[14] previous work focused on the removal of sulfur compounds from cyclic
structures in diesel at elevated temperature, the mechanism of sulfur
[15] removal using nanostructured reactive adsorbent can be potentially
extended to remove HS− ions under ambient conditions of anaerobic
[16]
digestions processes.
The primary goal of this study is to evaluate the suitability and
performance of ZnO NWs as an in situ reactive adsorbent of soluble
sulfides formed during the AD process. In the first part of the study, the
ability of the ZnO NWs to adsorb sulfides was tested in a model system
consisting of aqueous solution of sodium sulfide (Na2S). Sodium sulfide
is primarily used in the pulp and paper industry in the Kraft process. In
water at neutral pH, Na2S hydrolyzes to yield hydroxide and hydro-
sulfide ions (Eq. (4)).
Na2S + H2O ⇄ 2Na+ OH− + HS− (4)
Our approach is to use Na2S in water as a controlled source of HS−
and monitor the level of sulfide in water in the presence of the na-
nostructured adsorbent. This test will be a validation for the adsorbent’s
capability to remove sulfides. Following this validation, we propose to
test the adsorbent in situ during the AD process at varying sulfate levels
in the wastewater.
3. Materials and methods
3.1. Materials
Calcium chloride, magnesium chloride, ammonium chloride, po-
tassium phosphate monobasic, sodium sulfate were used as minerals
and nutrients for anaerobic digestion tests and were purchased from
Sigma-Aldrich. Sodium bicarbonate (Sigma-Aldrich) was used to adjust
alkalinity. A proprietary inorganic salt mix (Respirometer Systems &
Applications LLC, Springdale, AR, USA) was used as a source of trace
elements. Sodium sulfide nanohydrate was purchased from Sigma-
Aldrich and was used as a controlled source of HS− ions. Sodium hy-
droxide and hydrochloric acid were purchased from Sigma-Aldrich and
were used for pH adjustment. Ethanol was purchased from Sigma-
Aldrich and was used as a model source of soluble COD.
Granular anaerobic sludge was kindly provided by Anheuser-Busch
(St. Louis, MO). The sludge was a source for natural mixed anaerobic
microbial community from an operating full-scale anaerobic digester
treating brewery wastewater. The concentration of the bacteria in the
sludge was measured as Volatile Suspended Solids (VSS) content and
was determined to be 52.0 g/L. ZnO nanowires and ZnO nanowires
with NiO particles were obtained from Advanced Energy Materials LLC
− AdEM (Louisville, KY). AdEM produces the adsorbent using methods
discussed in [20].
3.2. Analytical methods
(2) Sulfide content in test solutions was determined colorimetrically
(3)
Company, Germany). Chemical analysis of sulfates was performed
spectrophotometrically using commercial test kits and DR 3900
Spectrophotometer (Hach Company, Germany). Gas analysis was per-
formed on SRI 8610C Gas Chromatograph (SRI Instruments Inc., Las
Vegas NV) using HayeSep D column (Restek Corporation) and thermal
conductivity detector (TCD) for methane and carbon dioxide detection;
MXT-1 column (Restek Corporation) and flame photometric detector
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R. Lupitskyy et al.
(FPD) was used for hydrogen sulfide detection. Scanning electron mi-
crographs and EDS spectra were acquired using NOVA NanoSEM 600
scanning electron microscope (FEI Company; Hillsboro, OR). X-ray
diffraction spectra were recorded using Discovery D-8 x-ray dif-
fractometer (Bruker; Billerica, MA). The identification of the crystal
phases was performed using the Powder Diffraction File (PDF) struc-
tural database.
3.3. Sulfide removal tests
Both ZnO NWs and ZnO NWs with NiO were used in the amount of
1 g/L and the concentration of sodium sulfide was varied from 100 to
1000 mg/L S2−. The sulfide removal tests were carried out for 15 h
(overnight) at room temperature under mechanical stirring.
Subsequently, the solution was filtered and the amount of sulfides in the
solution was determined colorimetrically using a commercial sulfide
kit. The ZnO NWs were recovered by filtration, dried in the oven at 60 C
and characterized using Energy-dispersive X-ray spectroscopy (EDS)
used in conjunction with scanning electron microscopy (SEM) and X-
ray diffraction (XRD).
3.4. Batch AD tests
Experimental set-up for laboratory-scale batch anaerobic digestion
tests was acquired from Respirometer Systems & Applications LLC,
Springdale, AR, USA. The system consists of a water bath placed on an
8-position magnetic stir plate, external pump, temperature controller,
and a pulse flow respirometer PF-800. Glass bottles of 500 ml were used
as reactors. Trace elements, minerals, nutrients, and NaHCO3 were
added to each bottle as described elsewhere [4,5]. As mentioned ear-
lier, the anaerobic sludge used in the AD tests was a natural mixed
anaerobic microbial community from an operating full-scale anaerobic
digester treating brewery wastewater at Anheuser-Busch (St. Louis,
MO).
For anaerobic digestion lab and pilot tests in our lab, our approach
has been to provide a “suspended solids free” waste water feed con-
taining only soluble COD. We have been using a model wastewater
prepared in the lab free of suspended solids and with a known chemical
composition for soluble form of COD during our lab scale batch AD
experiments and using waste water from a brewery during pilot scale,
continuous AD testing. In the present study, ethanol was used as a so-
luble substrate in the quantity of 6000 mg COD/L. In our past work, we
used a similar ethanol model feed for batch lab tests and successfully
scaled up the AD process to pilot scale for waste waters from a brewery
and a soy protein plant. The use of a model wastewater allows for
precise control of the substrate composition and properties required for
repeatable studies and is a common practice in batch AD experiments
[21–25].
Solution volume in each bottle was 400 ml. Granular anaerobic
sludge was added in the quantity of 12.0 g/L VSS. The pH after adding
the biomass, the substrate, and the nutrients was 7. The bottles were
purged with nitrogen for 1 min to ensure anaerobic conditions and
hermetically sealed with septum screw caps. The anaerobic digestion
tests were conducted under mesophilic conditions (35 °C). The volume
of the biogas produced was measured and recorded by the pulse flow
respirometer.
The anaerobic sludge was first stabilized by incubation with ethanol
as a substrate in a basal medium in order to insure an optimal and
steady microbial activity. The ethanol was introduced in a batch mode
and the gas production was monitored. When the gas production
stopped, the next batch of ethanol was introduced. A period of time
between the substrate (ethanol) loadings constitutes a feeding cycle.
The stabilization was conducted until the amount of gas produced was
repeatable from one feeding cycle to another. After the stabilization
was completed, the in-situ performance of the ZnO NWs during anae-
robic digestion process was evaluated. For the in situ use, the ZnO NW
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Journal of Environmental Chemical Engineering 6 (2018) 110–118
were prepared in the form of extrudates of approximately 2 mm dia-
meter using a mixture of 70% ZnO NW and 30% γ-Al2O3. The alumina
was added to improve the mechanical properties of the extrudates. The
extrudates in the amount of 1 g/L (based on ZnO) were placed in a
water-permeable pouch (mesh 200, nylon) for easy recovery and in-
troduced into the AD medium. Sulfates were added in the form of
Na2SO4 to make 2 g/L SO42−. The experiment was carried out for 3
feeding cycles. Sulfates were added at the beginning of each feeding
cycle. The gas produced during the digestion was collected into sample
bags and its composition was analyzed using gas chromatography.
4. Results and discussion
4.1. Sulfide removal in model system
According to Eq. (4), the dissociation of Na2S in water produces
HS− species which are the same species produced during the AD pro-
cess by the sulfate reducing bacteria. Therefore, in this study, the ability
of ZnO NWs to remove sulfides in an aqueous medium was first tested
using a model system composed of DI water and sodium sulfide. In-
itially, one test was conducted using ZnO NWs with and without NiO to
determine whether Ni was needed in the same manner as reactive ad-
sorbents used in hydrodesulfurization of diesel. Both ZnO NWs with and
without Ni removed sulfides from the solution to undetectable levels
after the treatment for 15 h. All the remaining tests were done using
ZnO NWs and the corresponding results are given in Table 2.
The results shown in Table 2 indicate that ZnO NWs can effectively
remove sulfides from aqueous solutions. In these tests, the initial sulfide
concentrations were arbitrarily chosen at 108 mg/L, 325 mg/L and
975 mg/L. At these concentrations, the resulting pH was about 12 due
to formation of OH− ions upon hydrolytic dissociation of Na2S (Eq.
(4)). At a ZnO dosage of 1 g/L and with overnight mixing (15 h), the
adsorbent reduced the final sulfate concentration to non- detectable
levels for the first two concentration values. For the 975 mg/L S2− level
only about two thirds of the sulfides were removed over 15 h giving a
maximum adsorbent capacity of 625 mg of S2− per gram of ZnO NW
adsorbent over 15 h of time − an average of 42 mg S2− per g ZnO per
hour. The efficacy of the adsorbent for S2− was also confirmed at
neutral pH − a typical pH level for anaerobic digestion (Test #2). It
should be noted that the pH of the solution did not change significantly
upon the sulfide removal. According to Eq. (2), hydroxyl ions released
during the reaction should further increase the pH of the solution. No
change in pH during the reaction points to a possible complexation of
hydroxyl ions with ZnS, which has been previously observed in aqueous
ZnS systems [26]. However, the incubation time seems to be a limiting
factor in this
process. This was observed in a test conducted for 1 h (#3) where less
than 10% of the sulfides out of 325 mg/L were removed. The predicted
mechanism upon which this system works is a metathesis, which depends
on the dissociation of ions in solution to proceed. The inability of the
system to remove the HS− ions in the shorter amount of time could be due
to several factors. One would be the lower solubility of ZnO in water.
Another possible explanation could be the presence of the NaOH from the
Table 2
Sulfide removal using ZnO NWs from aqueous Na2S solution.
Test Sulfides, mg/L S2− ZnO:Na2S molar Duration, hrs. pH
ratio
Initial Final Initial Final
1 108 n/da
3.6:1 15 11.3 11.3
2 325 n/da 1.2:1 15 7.5 7.6
3 325 300 1.2:1 1 11.7 11.9
4 325 n/da 1.2:1 15 11.7 11.9
5 975 350 0.4:1 15 12.2 12.3
a
Detection limit 0.03 mg/L.
R. Lupitskyy et al. Journal of Environmental Chemical Engineering 6 (2018) 110–118

Fig. 1. EDS spectra of the ZnO NWs: (A) original; (B) after incubation with 325 g/L S2− for 15 h (test #4, Table 2); (C) after incubation with 975 mg/L S2− for 15 h (test #5, Table 2).

dissociation of the Na2S in water. This hydroxide species could inhibit the
ability of the NaHS to react with the ZnO, or help inhibit the dissociation
of the ZnO, limiting its already low solubility. However, in an anaerobic
system where sulfate consuming bacteria produce soluble sulfides, the
inhibition from Na ions can be minimal.
The ZnO NWs before and after the tests #4 and #5 were analyzed by
Energy Dispersive X-ray Spectroscopy (EDS) (Fig. 1). X-ray diffraction
(XRD) of samples from tests #4 and #5 were included in Fig. 2. Ele-
mental analysis of the original ZnO NWs using EDS is shown in Fig. 1A
where no sulfur is present. However, the solid residues recovered after

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Fig. 2. X-Ray diffractograms of the ZnO NWs: (A)

original; (B) after incubation with 325 g/L S2− (test
#4, Table 2); (C) after incubation with 975 g/L S2−
(test #5, Table 2).

overnight treatment with 325 mg/L S2− and 975 mg/L S2− (Fig. 1B and
C respectively) clearly contained sulfur. Due to the inhomogeneity of Table 3
the solid residues, quantitative analysis using EDS is not reliable. Thus, H2S content in biogas during treatment of synthetic wastewater at three different sulfate
based on EDS spectra, it is not possible to make any conclusions about levels with and without ZnO NWs added in situ.

the relative amount of sulfur in the sample used with 325 mg/L S2−
Sulfates, mg/L 1900 3800 6500
(Fig. 1B), compared to the sample treated with 975 mg/L S2− (Fig. 1C).
A structural analysis using XRD (Fig. 2) showed a peak at 2θ value ZnO NWs No Yes No Yes No Yes
of 28.2 in the samples incubated with Na2S, which corresponds to zinc
Gas composition, % wt. CH4 68 68.7 67.3 67 65.1 65.2
sulfide. The figure showing the expanded spectra between 20 and 40
CO2 27.3 26.6 28.6 27.9 29.9 29.7
values of 2θ shows a small relative increase in the intensity of ZnS peak H2S 1.2 n/da 1.6 n/da 2 n/da
from the sample incubated with 325 mg/L S2− to the sample incubated
with 975 mg/L S2−. a
< 1 mg/L.

4.2. ZnO NWs testing in AD process

Following the evaluation using a model Na2S system, the ability of
ZnO NWs to capture soluble sulfides and prevent H2S formation has
been further investigated in-situ during the anaerobic digestion process.
The anaerobic sludge used in the AD tests was a natural mixed anae-
robic microbial community from an operating full-scale anaerobic di-
gester treating brewery wastewater and the other hand, the substrate
used in our experiments was a model artificial wastewater prepared in
the lab with known chemical composition. The use of a model waste-
water allows for precise control of the substrate composition and
properties required for repeatable studies and is a common practice in
batch AD experiments [21–25]. Our approach is to use the model waste
water in the lab scale batch AD experiments and use waste water from a
brewery during pilot scale, continuous AD testing. In our past work, we
used an ethanol model feed for batch lab tests and successfully scaled
up the AD process at pilot scale using waste waters from a brewery and
a soy protein plant. First, the efficiency of the ZnO NWs was evaluated
during the AD process at three levels of sulfate concentration in the
medium: 1900 mg/L, 3800 mg/L, and 6500 mg/L. The results are given
in Table 3, where it can be observed that upon the addition of the ZnO
NWs, H2S formation during the AD process decreased to a non-detect-
able level in all three tests. These tests validate our concept that the
hybrid approach proposed here has a strong potential to minimize H2S
formation during AD processing of sulfate containing waste water. This
inhibition of H2S formation, as discussed previously, can then lead to an
increase in the production of biogas and more importantly can protect
biomass from the toxic effect of soluble sulfides that are formed during
the AD treatment of waste waters containing high levels of sulfates.
In the second experiment, a longer-term performance of ZnO NWs
was studied for three consecutive feeding cycles. The sulfates in the
amount of 2.0 g/L were introduced at the beginning of each feeding
cycle along with the substrate (ethanol). The biogas production was
continuously monitored for the first two feeding cycles (Fig. 3, red and
green line for 1st and 2nd feeding cycle, respectively). A control test
was carried out to determine if ZnO NWs exhibit any toxicity to me-
thanogens (Fig. 3A) and compared with a control (batch containing no
ZnO NWs and no sulfates, blue line in Fig. 3). It can be seen that for two
feeding cycles there is no adverse effect of ZnO NWs on the methano-
genic activity. Another control test was carried out where sulfates were
added before each feeding cycle in the amount of 2.0 g/L SO42− to a
batch containing no ZnO NWs (Fig. 3B). This test simulates an anae-
robic treatment of a wastewater containing 2.0 g/L SO42−. Our pre-
liminary experiments with AD treatment of high sulfate waste streams
showed that this amount of sulfates is enough to cause a toxic effect
during a longer run. As can be seen from the biogas production graph,
no decrease in the methanogenic activity was observed after the first
feeding cycle. However, after the second feeding cycle, an 18% de-
crease in biogas production was observed compared to the first feeding
cycle and the control. This decrease indicates accumulation of dissolved
sulfides produced by sulfate-reducing bacteria, which has a toxic effect
on the methanogens. However, when sulfates were added to the batch
containing ZnO NWs (Fig. 3C), no significant decrease in biogas

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Table 4
In situ performance of ZnO NWs during three consecutive feeding cycles.
Feeding cycle Sulfates added, g/L SO42− H2S, % wt.
Control
1 2.0 1.0
2 2.0 1.18
3 2.0 2.29
production was observed. This shows that the application of reactive
adsorbent, such as ZnO NWs, in situ helps to reduce the sulfide toxicity
during the AD process.
The amount of H2S in the biogas was measured after each feeding
cycle using GC and the results are presented in Table 4. After the first
feeding cycle, the amount of H2S in the biogas was 1.0% wt. in the
control batch, whereas in the batch containing ZnO NWs, the amount of
H2S in the biogas was below the detection limit. During the second
feeding cycle, about 98% of the H2S was removed by the ZnO NWs
compared to the control batch. During the third feeding cycle, the
performance of the ZnO NWs started to decline resulting in the removal
of the 88% of the H2S compared to the control batch.
After the AD process, the ZnO NWs were recovered and analyzed
using EDS and XRD. Three samples were analyzed: 1) ZnO-γ-Al2O3
extrudates prior to AD process; 2) ZnO-γ-Al2O3 recovered after 3
feeding cycles from the control batch containing no sulfates; 3) ZnO-γ-
Al2O3 recovered after 3 feeding cycles from the batch where sulfates
were added. Elemental analysis of the Sample 1 using EDS (Fig. 4A)
showed presence of Zn, Al, and O indicating pure ZnO-γ-Al2O3. Sample
2 recovered after the AD process from the control batch showed a small
amount of phosphorus, which is most likely coming from the phos-
phates used as nutrients during the AD process (Fig. 4B). Sample 3
recovered after the AD process from the batch containing sulfates
showed small amounts of phosphorus and calcium, which were present
Journal of Environmental Chemical Engineering 6 (2018) 110–118
Fig. 3. Biogas production during batch anaerobic
digestion process: A) ZnO NWs only, no sulfates
added; B) sulfates added, no ZnO NWs; C) ZnO NWs
and sulfates added.
in the AD medium, as well as a significant amount of sulfur, indicating
that the H2S was indeed adsorbed by the ZnO-γ-Al2O3 (Fig. 4C). Ana-
lysis of the ZnO-γ-Al2O3 samples using XRD (Fig. 5) showed the ZnS
formation in the sample recovered from the batch containing sulfates
(Fig. 4C). The three diffraction peaks at 2θ values of 28.2, 47.1, and
ZnO NWs
56.1 correspond to cubic ZnS. Broadening of the XRD peaks suggest the
n/d polycrystalline nature of ZnS.
0.025
0.27
4.3. Future study and proposed system configuration
It has been demonstrated at lab scale that the proposed approach to
prevent H2S formation in biogas in situ has a strong potential to reduce
extensive post-processing costs. The AD tests were done in a batch
mode using a controlled model feed solution containing ethanol.
Further experiments will involve a long-term performance evaluation of
the developed reactive adsorbent during a continuous AD operation
using brewery waste water to verify the consistency of performance,
allowing for better economic analysis regarding potential for power
grid use. A proposed system configuration will be a packed bed reactor
that uses cartridges of said nanowires that can be easily inserted, re-
generated, and replaced. The life cycle, capacity, and regeneration
ability of the ZnO NWs, as well as potential long-term toxicity to
anaerobic microorganisms will also be investigated. Ultimately, the
process and reactor design could be optimized to produce a maximum
amount of biogas for utilization.
5. Conclusions
In this work, ZnO nanowires have been evaluated as a reactive ad-
sorbent for in situ removal of soluble sulfides formed by anaerobic bacteria
and prevention of H2S formation in biogas during anaerobic digestion. The
nanostructured reactive adsorbent has been shown to be effective in the
removal of soluble sulfides first in the model solution of sodium sulfide
and then in situ during the anaerobic digestion process, preventing for-
mation of H2S in biogas. Compared to reactive adsorbents used for
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Fig. 4. EDS spectra of the ZnO-γAl2O3 extrudates: (A) original; (B) after AD process without sulfates; (C) after AD process with sulfates.

hydrodesulphurization at elevated temperatures, the current approach
removed sulfides at room temperature. It has also been found that ZnO
NWs have the capacity to prevent sulfide-induced toxicity to methanogens
when treating a wastewater containing high level of sulfates (2.0 g/L).
Based on the results and the fact that ZnO is a sorbent that can be
regenerated, the proposed method has a potential to become an effective,
low-cost, robust, and sustainable technology for removal of H2S from
biogas, as well as prevention of sulfide-induced toxicity to methanogenic
microorganisms, thus allowing for digestion of waste streams with high
sulfate content without the need for any pretreatment.

116
R. Lupitskyy et al.
Acknowledgements
This work is partly supported by a generous gift from Brown-
Forman Company, Louisville, KY. The authors would like to thank
Advanced Energy Materials LLC for the supply of nanomaterials and
Anheuser-Busch for the supply of AD sludge. We also want to thank Dr.
Mahendra Sunkara of Conn Center for valuable suggestions during the
study.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in the
online version, at https://doi.org/10.1016/j.jece.2017.11.048.
References
[1] J. Bekkering, A.A. Broekhuis, W.J. van Gemert, Optimisation of a green gas supply
chain–a review, Bioresour. Technol. 101 (2010) 450–456.
[2] F. Tambone, P. Genevini, G. D'Imporzano, F. Adani, Assessing amendment prop-
erties of digestate by studying the organic matter composition and the degree of
biological stability during the anaerobic digestion of the organic fraction of MSW,
Bioresour. Technol. 100 (2009) 3140–3142.
[3] T. Rehl, J. Muller, Life cycle assessment of biogas digestate processing technologies,
Resour. Conserv. Recy. 56 (2011) 92–104.
[4] W. Charles, R. Cord-Ruwisch, G. Ho, M. Costa, P. Spencer, Solutions to a combined
problem of excessive hydrogen sulfide in biogas and struvite scaling, Water Sci.
Technol. 53 (2006) 203–210.
[5] J.E. Burgess, S.A. Parsons, R.M. Stuetz, Developments in odour control and waste
gas treatment biotechnology: a review, Biotechnol. Adv. 19 (2001) 35–63.
[6] H.Q. Ge, L.S. Zhang, D.J. Batstone, J. Keller, Z.G. Yuan, Impact of iron salt dosage to
sewers on downstream anaerobic sludge digesters: sulfide control and methane
production, J. Environ. Eng.-ASCE 139 (2013) 594–601.
[7] J. Kanjanarong, B.S. Giri, D.P. Jaisi, F.R. Oliveira, P. Boonsawang, S. Chaiprapat,
R.S. Singh, A. Balakrishna, S.K. Khanal, Removal of hydrogen sulfide generated
117
Journal of Environmental Chemical Engineering 6 (2018) 110–118
Fig. 5. X-Ray diffractograms of the ZnO-γAl2O3 ex-
trudates: (A) original; (B) after AD process without
sulfates; (C) after AD process with sulfates.
during anaerobic treatment of sulfate-laden wastewater using biochar: evaluation of
efficiency and mechanisms, Bioresour. Technol. 234 (2017) 115–121.
[8] E.L. Barrera, H. Spanjers, O. Romero, E. Rosa, J. Dewulf, Characterization of the
sulfate reduction process in the anaerobic digestion of a very high strength and
sulfate rich vinasse, Chem. Eng. J. 248 (2014) 383–393.
[9] J. Li, L. Yu, D.S. Yu, D. Wang, P.Y. Zhang, Z.G. Ji, Performance and granulation in
an upflow anaerobic sludge blanket (UASB) reactor treating saline sulfate waste-
water, Biodegradation 25 (2014) 127–136.
[10] J.A. Oleszkiewicz, T. Marstaller, D.M. Mccartney, Effects of ph on sulfide toxicity to
anaerobic processes, Environ. Technol. Lett. 10 (1989) 815–822.
[11] B.R. Dhar, E. Youssef, G. Nakhla, M.B. Ray, Pretreatment of municipal waste acti-
vated sludge for volatile sulfur compounds control in anaerobic digestion,
Bioresour. Technol. 102 (2011) 3776–3782.
[12] S.K. Khanal, J.C. Huang, Online oxygen control for sulfide oxidation in anaerobic
treatment of high-sulfate wastewater, Water Environ. Res. 78 (2006) 397–408.
[13] E.B. de Jesus, L.R.P.D. Lima, Simulation of the inhibition of microbial sulfate re-
duction in a two-compartment upflow bioreactor subjected to molybdate injection,
Bioprocess Biosyst. Eng. 39 (2016) 1201–1211.
[14] L. Krayzelova, J. Bartacek, I. Diaz, D. Jeison, E.I.P. Volcke, P. Jenicek,
Microaeration for hydrogen sulfide removal during anaerobic treatment: a review,
Rev. Environ. Sci. Biol. 14 (2015) 703–725.
[15] P.F. Henshaw, W. Zhu, Biological conversion of hydrogen sulphide to elemental
sulphur in a fixed-film continuous flow photo-reactor, Water Res. 35 (2001)
3605–3610.
[16] K.Y. Show, D.J. Lee, X.L. Pan, Simultaneous biological removal of nitrogen-sulfur-
carbon: recent advances and challenges, Biotechnol. Adv. 31 (2013) 409–420.
[17] L. Zhang, P. De Schryver, B. De Gusseme, W. De Muynck, N. Boon, W. Verstraete,
Chemical and biological technologies for hydrogen sulfide emission control in
sewer systems: a review, Water Res. 42 (2008) 1–12.
[18] B.L. Dou, C. Wang, H.S. Chen, Y.C. Song, B.Z. Xie, Y.J. Xu, C.Q. Tan, Research
progress of hot gas filtration, desulphurization and HCl removal in coal-derived fuel
gas: a review, Chem. Eng. Res. Des. 90 (2012) 1901–1917.
[19] I. Rosso, C. Galletti, M. Bizzi, G. Saracco, V. Specchia, Zinc oxide sorbents for the
removal of hydrogen sulfide from Syngas, Ind. Eng. Chem. Res. 42 (2003)
1688–1697.
[20] F.G. Petzold, J. Jasinski, E.L. Clark, J.H. Kim, J. Absher, H. Toufar, M.K. Sunkara,
Nickel supported on zinc oxide nanowires as advanced hydrodesulfurization cata-
lysts, Catal. Today 198 (2012) 219–227.
[21] H.Q. Yu, H.H.P. Fang, Anaerobic acidification of a synthetic wastewater in batch
R. Lupitskyy et al.
reactors at 55 °C, Water Sci. Technol. 46 (2002) 153–157.
[22] G. Akila, T.S. Chandra, Performance of an UASB reactor treating synthetic waste-
water at low-temperature using cold-adapted seed slurry, Process Biochem. 42
(2007) 466–471.
[23] Y.F. Zhang, I. Angelidaki, Counteracting ammonia inhibition during anaerobic di-
gestion by recovery using submersible microbial desalination cell, Biotechnol.
Bioeng. 112 (2015) 1478–1482.
[24] M. Shakeri, M. Vossoughi, Coupled anaerobic baffled reactor (ABR)/Activated
sludge treatment of synthetic waste water with high concentration of sulfate and
118
Journal of Environmental Chemical Engineering 6 (2018) 110–118
COD, IJE Trans. B: Appl. 16 (2003) 11–20.
[25] J. Sun, X. Dai, Q. Wang, Y. Pan, B.-J. Ni, Modelling methane production and sulfate
reduction in anaerobic granular sludge reactor with ethanol as electron donor, Sci.
Rep. 6 (2016) 35312 | 35310.31038/srep35312.
[26] B.R. Tagirov, O.M. Suleimenov, T.M. Seward, Zinc complexation in aqueous sulfide
solutions: determination of the stoichiometry and stability of complexes via ZnS(cr)
solubility measurements at 100 °C and 150 bars, Geochim. Cosmochim. Acta 71
(2007) 4942–4953.