See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/281324526

Synergistic effect of alkaline pretreatment and Fe dosing on batch anaerobic

digestion of maize straw

Article  in  Applied Energy · November 2015

DOI: 10.1016/j.apenergy.2015.08.045

CITATIONS READS

29 637

6 authors, including:

Shailendra Khatri Shubiao Wu

Nepal Agricultural Research Council China Agricultural University & Aarhus University
2 PUBLICATIONS   30 CITATIONS    108 PUBLICATIONS   1,604 CITATIONS   

SEE PROFILE SEE PROFILE

Simon Kizito Zhang Wanqin

Makerere University China Agricultural University
23 PUBLICATIONS   300 CITATIONS    20 PUBLICATIONS   402 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Msc Thesis View project

Biomass gasifier stove View project

All content following this page was uploaded by Simon Kizito on 16 March 2018.

The user has requested enhancement of the downloaded file.

 Applied Energy 158 (2015) 55–64

Contents lists available at ScienceDirect

Applied Energy
journal homepage: www.elsevier.com/locate/apenergy

Synergistic effect of alkaline pretreatment and Fe dosing on batch

anaerobic digestion of maize straw
Shailendra Khatri a,b, Shubiao Wu a,⇑, Simon Kizito a,c, Wanqin Zhang a, Jiaxi Li a, Renjie Dong a
a
Key Laboratory for Clean Renewable Energy Utilization Technology of Ministry of Agriculture, College of Engineering, China Agricultural University, Beijing 100083, People’s
Republic of China
b
Agricultural Engineering Division (AED), Nepal Agricultural Research Council (NARC), Khumaltar, Lalitpur, Bagmati, Nepal
c
College of Agriculture and Environmental Sciences, Makerere University, PO Box 7062, Kampala, Uganda

h i g h l i g h t s g r a p h i c a l a b s t r a c t

 Synergistic effect of NaOH treatment

and Fe dosage to maize straw was
investigated.
 Combining NaOH treatment and Fe
dosing resulted in 57% and 56% higher
biogas and methane yield
respectively.
 Combined treatment shortened the
technical digestion time from 48 days
to 7 days.
 Methane content did not differ
significantly among the straw
treatments.

a r t i c l e i n f o a b s t r a c t

Article history: The synergistic effect of alkaline pretreatment and Fe dosing on anaerobic digestion of maize straw was
Received 21 December 2014 investigated using mesophilic batch reactors. Three straw treatments were investigated as follows: NaOH
Received in revised form 20 July 2015 (4% and 6%) pretreatment, Fe dosage (50, 200, 1000 and 2000 mg/L), and combined NaOH pretreatment
Accepted 15 August 2015
and Fe dosage. Compared to the control, NaOH pretreatment alone increased methane yield by 3.5%
(313.3 mL CH4/gVS) and 22.5% (370.9 mL CH4/gVS) and shortened the technical digestion time (TDT) from
48 days to 19 days and 10 days in 4% NaOH and 6% NaOH pretreatment respectively. Moreover, Fe dosing
Keywords:
(200–1000 mg/L) alone gave a methane yield higher (9.4%) than that obtained from 4% NaOH and 7.5%
Anaerobic digestion
Maize straw
less than the methane yield from 6% NaOH pretreatment; however, the TDT was 10 days longer.
NaOH pretreatment Combining NaOH pretreatment and Fe dosage (200–1000 mg/L) significantly increased the methane yield
Fe dosing even further to 21.8% (368.8 mL CH4/gVS) and 56.2% (472.9 mL CH4/gVS), and shortened TDT from
Synergistic effect 48 days to 13 days and 7 days in 4% NaOH and 6% NaOH pretreatment respectively. This synergistic effect
may be attributed to the fact that the alkaline treatment improved accessibility of the biodegradable frac-
tion of the straw while Fe contributed to increased microbial enzyme activity.
Ó 2015 Elsevier Ltd. All rights reserved.

⇑ Corresponding author. Tel.: +86 10 62737852; fax: +86 10 62736067.

E-mail address: wushubiao@gmail.com (S. Wu).

http://dx.doi.org/10.1016/j.apenergy.2015.08.045
0306-2619/Ó 2015 Elsevier Ltd. All rights reserved.
56 S. Khatri et al. / Applied Energy 158 (2015) 55–64
1. Introduction
Around the world, increasing population and industrialization
is heightening the energy demand. Given the slow development
of alternative energy sources especially for growing economies,
the energy demand is by far based on huge consumption of fossils
fuel i.e., coal, petroleum and natural gas. For instance, China’s
dependence on fossil oil is about 46.6% for industrial and infras-
tructure development sectors [1]. As a result, overconsumption of
fossil fuels is causing heavy pollution releasing green house gases
(GHGs). Biogas technology is becoming the means of reducing
energy poverty that is considered as a serious barrier to sustainable
economic development in developing countries. In China, the bio-
gas infrastructure has been highly prioritized by the government as
an eco-friendly and sustainable forms of renewable energy. How-
ever, biogas production is mostly based on animal manures,
municipal wastes and sewage sludge and little from lignocellulosic
biomass [2,3]. Given that theoretically lignocellulosic biomass pos-
sess a high bioenergy (biomethane and bioethanol) potential than
animal manures and sewage sludge, their integration into the pro-
duction cycle could improve producible energy amount.
The use of lignocellulosic biomass such as wheat, rice, bagasse,
and maize straw is increasingly recognized as the future feedstock
that will enable sustainable bioenergy production. Abundance, low
material cost, and high energy content make lignocellulosic bio-
mass the preferred feedstock [4]. Lignocellulosic biomass produces
both a clean biogas fuel as well as bioenergy residues that can be
used as nutrient rich fertilizer, utilizing some of the energy of the
chemical bonds within the biomass [5]. Importantly, the use of
these agricultural residues in anaerobic digestion reduces the
release of environmentally detrimental carbon dioxide and other
greenhouse gases into the atmosphere. Moreover, researchers have
proven that the use of lignocellulosic biomass in biogas production
gives a high biogas yield with a higher CH4 concentration than that
of biogas typically produced with animal manures [6].
In China, over 600 million tons of crop straw residues, including
250 million tons from maize straw alone, are produced every year
[7,8]. These residues could be of great potential for bioenergy pro-
duction, especially for biogas and bio-ethanol. However, the full
integration of straw biomass into biogas production is still very
slow [1]. The major obstacle for producing biogas from straw has
been attributed to the complex lignin structure that presents a
huge barrier for anaerobic bacteria to perform holocellulose
hydrolysis. Because of this barrier in hydrolysis, anaerobic fermen-
tation of straw results in a long lag phase prior to gas production
and consequently, long retention times for the eventual anaerobic
degradation.
To improve the biodegradability of straw, much research has
been devoted to co-digestion of the lignocellulosic biomass with
other substrates such as manure and sludge [7,9]. Previous studies
have used co-digestion of straw with chicken manure [10], cow
dung [9,11], pig manure [12] and waste activated sludge [13,14].
From all these mentioned studies, the biogas yield from co-
digestion has been reported to be higher than that from the
mono-digestion of the lignocellulosic biomass; however, it is still
below the potential theoretical chemical energy of the biomass
[7,15].
Currently, research is progressing toward pre-digestion biomass
treatments with the goal of breaking down lignin and exposing
holocellulose to allow for more microbial degradation. The most
widely applied pretreatments are physical, chemical, thermal and
biological as well as combinations of these methods [16–18]. Phys-
ical methods such as grinding, milling, extrusion and irradiation
have received reasonable attention in research [19,20]. Biologi-
cally, fungal (white and brown rot), bacteria pretreatments, and
enzymatic ensilaging have been widely studied [4,18,21]. As for
the chemical method, alkaline pretreatments using strong bases
such as sodium hydroxide, calcium oxide, and peroxides have been
well studied in addition to the use of acids such as sulfuric, acetic,
and nitric acids [16,20,22]. Microwave, steam explosion, and liquid
hot water have dominated thermal pretreatments [23].
Compared to purely biological or physical treatments, the com-
bination of physiochemical and thermochemical pretreatments are
regarded as promising methods for industrial application because
of their superb ability to breakdown the non-digestible lignin.
Alternatively, for small scale pretreatments, alkaline NaOH pre-
treatment has been reported as one of the most effective methods,
and has other positive outcomes like increase of accessible surface
area, decrease in degree of holocellulose polymerization and crys-
tallinity, and disruption of lignin structure [17,24,25].
Besides overcoming the lignin barrier, another important aspect
of anaerobic digestion of straws is the nutrient supply to the
microbes within the reactor. Authors of recent reviews partially
attribute the poor efficiency of biogas production from plant-
based straws/residues to the deficiency of trace elements in the
digestion liquor that treats lignocellulose biomass [26–28].
Microorganisms involved in anaerobic biomass fermentation
require carbon, nitrogen and trace elements for maximum gas
yield and proper process stability. Lignocellulosic biomass, includ-
ing maize straw, usually has adequate carbon but is lacking in
some essential trace elements. Thus, anaerobic digestion requires
supplementation for effective microbial activity and metabolism
[26,29]. The requirements of trace elements vary strongly by the
metal species and its bioavailability to microorganisms [30]. Essen-
tial metals, including Fe, Mo, Ni, Co, Mn, W and Se, are important in
both the microbial enzyme system and microbial respiration pro-
cesses as cofactors and extracellular electron acceptors, respec-
tively [31–33].
In this study, we hypothesized that iron (Fe) is one of the essen-
tial trace elements that can stimulate biogas production from
maize straw. Iron, primarily due to its redox properties, is an
important micro-nutrient for bacterial growth and is vital for elec-
tron transport via the iron-sulfur cluster and activity of various
enzymes, cytochromes, and co-enzymes (hydrogenase, CO-
dehydrogenase, NO-reductase, superoxide dismutase, nitrite and
nitrate reductase, and nitrogenase) [29,33,34].
Previous studies have reported the effects of trace elements and
the different pretreatments on anaerobic digestion of maize straw
and silage. For instance, Hinken et al. [31] performed Fe dosing
with maize silage and reported a 35% increase from the control
in biogas production at 205 mg/L Fe. Likewise, Song et al. [35],
investigated the effects of seven chemical pretreatments including
four acids; (H2SO4, HCl, H2O2, and CH3COOH) at concentrations of
1%, 2%, 3%, and 4% (w/w) and three alkaline reagents (NaOH, Ca
(OH)2, and NH3. H2O) at concentrations of 4%, 6%, 8%, and 10%
(w/w) on the methane yield of corn straw. From their results, the
straw pretreated with 8% NaOH acquired methane yield of
163.5 mL CH4 g/VS which was lower than that achieved in 3%
H2O2 (216.7 mL CH4 g/VS) and 8% Ca(OH)2 (206.6 mL CH4 g/VS).
In a similar study, Zhu et al. [25], applied different NaOH pretreat-
ments (1%, 2.5%, 5.0% and 7.5%) (w/w) to investigate their effect on
biogas production from corn stover. From their findings, the high-
est biogas yield of 372.4 L/kg VS was obtained with 5% NaOH-
pretreated corn stover, which was 37.0% higher than that of the
untreated corn stover. From our literature survey, we found no
studies addressing the potential synergistic effect of alkaline pre-
treatment and Fe dosing for enhanced biogas production.
To address this gap, this study evaluated technical digestion
time, biogas and methane yield, and methane content of the pro-
duced biogas to examine potential synergistic effects of NaOH pre-
 S. Khatri et al. / Applied Energy 158 (2015) 55–64 57

treatment and Fe dosing on anaerobic digestion of maize straw. We ments were conducted using 250 mL serum bottles with a working
also evaluated the effects of the combined NaOH pretreatment and volume of 180 mL. Reactor bottles were prepared by adding 10 g
varying Fe dosing on process-limiting parameters such as pH, VS/L inoculum (119 mL), feedstock at a feedstock to inoculum ratio
ammonium nitrogen (NH+4-N) and volatile fatty acids (VFAs) during of 1:1 (w/w) based on the volatile solids [20], and different Fe
anaerobic digestion of maize straw. Furthermore, preliminary eco- dosages, in that order. Finally, distilled water was added to a final
nomic feasibility evaluations and ecological impacts of proposed working volume of 180 mL. The maize straw groups tested were as
treatments were also integrated in this study. follows: no treatment (no NaOH or Fe dose), Fe dosed with no
NaOH, 4% NaOH pretreatment, 6% NaOH pretreatment, 4% NaOH
2. Materials and methods pretreatment dosed with Fe, and 6% NaOH pretreatment dosed
with Fe. All Fe dosed groups were tested at the concentrations of
50, 200, 1000 and 2000 mg/L. The initial pH of the mixed liquor
2.1. Inoculum
in each reactor bottle was adjusted to 7.0 ± 0.1 with addition of
1 M HCl or 1 M NaOH [36]. The headspace of each reactor bottle
The inoculum used in this study was taken from a lab-scale
was flushed with 100% pure N2 for 2 min in order to remove the
anaerobic digester that had been digesting chicken manure for
residual oxygen and ensure anaerobic conditions, and bottles were
two years at controlled temperature (37 ± 1 °C). Prior to use, the
inoculum was incubated at 37 ± 1 °C for 2 weeks to ensure degra- sealed with rubber stoppers. Assays with inoculum only were used
to determine the amount of biogas produced. This amount was
dation of residual organic matter and to remove the dissolved
methane [22]. Table 1 summarizes the inoculum characteristics later subtracted from the sample assays containing inoculum and
maize straw to calculate the biogas volume produced only from
after incubation.
the straw. All experiments were done in triplicate (48 trials in
total) and ran for 52 days. After 52 days, the residual digestate
2.2. Maize straw
was analyzed for volatile solids (VS), volatile fatty acid (VFA)/total
inorganic carbon (TIC) and NH+4-N.
Maize straw was collected from a farm owned by China Agricul-
tural University and located on university premises in Beijing,
China. The straw was air-dried for 30 days to less than 10% mois-
2.5. Analytical methods
ture content. Using a grinding mill, the air-dried straw was ground
to less than 1 mm particle size and stored at room temperature in
2.5.1. Chemical composition analysis
air-tight plastic bags. Portions of the ground straw were used for
Compositional analysis of maize straw for lignin, cellulose and
the following: proximate analysis, pretreatment with NaOH at 4%
hemicellulose was performed using an automatic cellulose ana-
and 6% concentration, and later, anaerobic digestion (i.e. no
lyzer (ANKOMA200i, USA). Ultimate analysis was used to deter-
pretreatment).
mine carbon, nitrogen and hydrogen content of dried maize
Alkaline pretreatment was done according to Chandra et al.
straw on an elemental analyzer (Vario EL/micro cube, Germany).
[22]. Briefly, 60 g of oven-dried ground maize straw was mixed
Proximate analysis of straw and digestate for total solids (TS), vola-
with 2.4 g and 3.6 g solid NaOH to achieve 4% and 6% NaOH (w/
tile solids (VS) and ammonium nitrogen were determined accord-
w) treatment, respectively. Water was added to achieve 10% total
ing to APHA standard method (2540G, 2540D, 4500-NH3D) [37].
solids concentration and after thorough agitation, the bottles were
The maize straw anaerobic digestion performance was assessed
sealed and incubated at 37 ± 1 °C for 5 days. At the end of the incu-
by gas production (rate and yield) and methane content of the bio-
bation period, the biomass was stored for proceeding experiments.
gas. Process performance was evaluated by periodic monitoring of
pH, production of volatile fatty acids (VFAs), and NH+4-N. Samples
2.3. Chemicals of 1.5 mL per bottle were withdrawn using gas tight syringes for
analyzing these parameters at every 3 days interval. The pH was
The stock Fe solution (10,000 mg/L) was prepared by dissolving measured directly using a digital pH meter (FE20, METTLER
35.6 g anhydrous FeCl2.4H2O solid (99% purity) in 1 L distilled TOLEDO, Switzerland) with a glass electrode (LE438, METTLER
water. Appropriate dilutions were made to achieve the desired TOLEDO Switzerland). For determination of NH+4-N concentration,
concentrations ranging from 50, 200, 1000, 2000 mg/L Fe. samples were centrifuged at 8000 rpm for 15 min at 4 °C, and the
supernatant was filtered through cellulose acetate membrane
2.4. Experimental set-up (0.45 lm). The TIC and VFA content were determined by titration
with 0.1 N H2SO4 to endpoints of pH 5.0 and 4.4 according to the
A schematic view of the experimental set-up designed for this Nordmann method [36].
study is shown in Fig. 1. Both the pretreated and untreated maize The single VFA concentration for acetic acid, propionic acid, and
straws were used directly without any further washing. Experi- butyric acid were determined by gas chromatography (Shimadzu,
GC-2010 Plus, Kyoto, Japan). A flame ionization detector and a
Rtx-wax capillary column (30 m  0.25 mm  0.25 lm) was used
Table 1
with high purity nitrogen as the carrier gas at a flow rate of
Characteristics of inoculum and maize straw used in the present study.
40 mL/min and a split ratio of 30. The temperature of the column
Parameters Inoculum Untreated 4% NaOH 6% NaOH began at 60 °C (2 min holding time). The column temperature
TS (%) 2.64 ± 0.24 94.37 ± 0.13 12.04 ± 0.10 11.42 ± 0.08 was increased at 10 °C/min to 140 °C, and then increased further
VS (%) 1.50 ± 0.10 85.50 ± 0.24 10.33 ± 0.07 9.52 ± 0.21 at 20 °C/min to 230 °C (5 min holding time). The temperatures of
VS/TS 57.37 ± 1.83 90.59 ± 0.12 85.79 ± 0.10 83.33 ± 1.49
the injector and detector were 230 °C and 250 °C, respectively.
pH 7.85 ± 0.01 – 6.38 ± 0.10 6.81 ± 0.20
Cellulose (%) – 38.33 ± 0.85 37.08 ± 2.74 37.01 ± 0.53
Samples for VFA analysis were prepared by filtering the super-
Hemicellulose (%) – 29.76 ± 1.35 15.00 ± 0.29 8.15 ± 0.29 natant of the centrifuged sample through 0.45 lm and subse-
Lignin (%) – 3.82 ± 0.53 3.42 ± 0.52 3.10 ± 0.21 quently, 0.22 lm cellulose acetate membranes. The obtained
L/C – 0.1 0.09 0.08 soluble filtrate was acidified using formic acid. VFA concentrations
All values are expressed as means ± standard deviation (n = 3). TS = total solids, were determined using a standard curve of acetic, propionic, buty-
VS = volatile solids, L/C = Lignin/cellulose. ric, valeric acid, and caproic acids.
58 S. Khatri et al. / Applied Energy 158 (2015) 55–64
Fig. 1. Schematic diagram of experimental set up for batch anaerobic digestion in this study.
2.5.2. Biogas production measurement and analysis
Daily biogas volume production was measured using the salted
water displacement method [36]. The methane content in biogas
samples were determined by gas chromatography (GC-2010 plus,
SHIMADZU, Japan). The gas chromatograph had a stainless steel
column of TDX-01 (packed with carbon molecular sieve,
2 m  3 mm) and a thermal conductivity detector (TCD). Nitrogen
was used as the carrier gas with a flow rate of 30 mL/min. The tem-
peratures of the TCD, injector, and column oven were 150 °C,
120 °C and 80 °C, respectively. A standard gas consisting 59.7%
CH4 and 40.3% CO2 (v/v) was used for the calibration of the system.
Gas samples (30 lL) for measurement of methane content were
withdrawn through a syringe from the headspace of each bottle.
The cumulative biogas production was calculated by adding each
daily biogas yield until 52 days.
2.6. Data analysis
Experiments were performed in triplicate and all chemical anal-
yses and measurements are displayed as average values of tripli-
cate readings. Descriptive analysis was done to summarize data
into averages, standard deviations, and standard error values. Anal-
ysis of Variance (Two-Way ANOVA) was used to investigate poten-
tial differences in biogas yield within and between the treatments.
All inferential analysis was done within the 95% level of confidence
(p = 0.05) using Sigma Plot software version 12.5 (Systat Software
Inc, USA).
3. Results and discussion
3.1. Effect of NaOH pretreatment on maize straw composition
Table 1 shows the characteristics of the maize straw and inocu-
lum used in the present study. Maize straw had TS, VS and VS/TS
ratio of 94.37%, 85.50%, and 90.59%, respectively. The high VS/TS
ratio proves that most solid components were organics, which
are appropriate for methane production. The composition analysis
showed cellulose and hemicellulose as the main carbon source,
68.09% of dry weight content of straw. The other portion of dry
weight content was lignin, crude fats and proteins, and hot water
extractives (sugar, starch, pectin, etc.).
Additionally, Table 1 depicts the changes in the lignocellulose
composition of straw resulting from NaOH pretreatment. Alkaline
pretreatment of straw only slightly solubilized lignin, but greatly
solubilized the hemicellulose fraction. Compared to the untreated
straw, 4% NaOH pretreated samples had 37.08%, 15.0%, 3.42% cellu-
lose, hemicellulose and lignin content, respectively while 6% NaOH
had 37.01%, 8.15%, 3.10%. The reduction in hemicellulose might be
due to the alkali attack on the hemicellulose and its subsequent
conversion to byproducts like hot water extractives [24]. Less cel-
lulose reduction indicated that NaOH might increase internal sur-
face area and break down the cellulosic crystalline areas for
easier biodegradation [17,20]. According to Scharer and Moo-
young [38], substrate biodegradability can be estimated by the lig-
nin (L)/cellulose (C) ratio. Low L/C ratio indicates a high degree of
digestibility. The L/C ratios of 4% and 6% NaOH pretreated (0.09,
0.08) were lower than the untreated (0.1) indicating enhanced
biodegradability through NaOH pretreatment.
3.2. Effect of NaOH pretreatment on biogas and methane yield
The daily biogas yield and cumulative biogas production during
52 days of anaerobic digestion are shown in Fig. 2. The biogas mea-
surement started at two days. Both the achieved maximum daily
biogas yield (Dmax) and cumulative biogas yield from pretreated
straw was greater than that from untreated straw (no NaOH). For
instance, Dmax of untreated straw was 52.6 mL/gVS, while Dmax of
4% and 6% NaOH pretreated straw was 72.8 mL/gVS and
 S. Khatri et al. / Applied Energy 158 (2015) 55–64 59

160 900
A Untreated B

Cumulave biogas yield (mL/gVS)

140 800

Daily biogas yield (mL/gVS-d)

700
120
600
100
500
80
400
60
300
40
200
20 100

0 0
160 900
C 4% NaOH pretreated D

Cumulave biogas yield (mL/gVS)

140 800
Daily biogas yield (mL/gVS-d)

700
120
600
100
500
80
400
60
300
40
200
20 100

0 0
160 900
E 6% NaOH pretreated F
800

Cumulave biogas yield (mL/gVS)

140
Daily biogas yield (mL/gVS-d)

700
120
600
100
500
80
400
60
300
40
200
20 100

0 0
0 5 10 15 20 25 30 35 40 45 50 55 0 5 10 15 20 25 30 35 40 45 50 55
Time (d) Time (d)
Fe 0 mg/L Fe 50 mg/L Fe 200 mg/L Fe 1000 mg/L Fe 2000 mg/L

Fig. 2. Daily biogas yield and cumulative biogas yield of untreated (no NaOH) (A and B), 4% NaOH pretreated (C and D) and 6% NaOH pretreated maize straw (E and F).

111.7 mL/gVS, respectively. Likewise, higher alkaline (6% NaOH)
pretreated straw had the highest cumulative biogas yield
(670.9 mL/gVS) over lower alkaline (4% NaOH) pretreated
(570.7 mL/gVS) and untreated straw (530.6 mL/gVS) (Table 2). A
higher gas yield could be a result of easier accessibility of the holo-
cellulose for microbial degradation. This easier accessibility is
afforded by the NaOH pretreatment that may have caused break-
down of the complex structure of holocellulose and separation of
lignin content.
Fig. 3 shows the daily methane yield (DMY) and cumulative
methane yield (CMY) obtained at different treatment conditions
during digestion of maize straw. For untreated straw, the maxi-
mum daily methane yield (Dmmax) was observed at 16 day
(28.8 mL CH4/gVS-day). However, for 4% and 6% NaOH pretreated
straw, Dmmax of 31.1 mL CH4/gVS-day and 61.8 mL CH4/gVS-day
were achieved on 3 day and 2 day respectively. The cumulative
average methane yields of untreated, 4% NaOH and 6% NaOH pre-
treated were 302.81, 313.35 and 370.99 mL CH4/gVS, respectively
(Fig. 3 and Table 2). The obtained cumulative methane yield for
6% NaOH was 22.5% and 19.0% higher than untreated and 4% NaOH
pretreated straw respectively. The earlier occurrence of daily
methane yield peaks coupled with improved methane production
rate and shorter effective methane production time for the pre-
treated straw could be due to effective destruction of lignin struc-
ture and thus easier microbial degradation of the holocellulose.
3.3. Effect of NaOH pretreatment on technical digestion time
In this study, technical digestion time (TDT) refers to the time
taken by different treatments to achieve 50% (T50), 80% (T80), and
100% (T100) value of total cumulative biogas yield for the pretreated
straw based on the control (no NaOH and Fe dosage). TDT is a mea-
60 S. Khatri et al. / Applied Energy 158 (2015) 55–64

Table 2
Comparison of anaerobic digestion performance between no NaOH, 4% NaOH and 6% NaOH pretreated maize straw at different Fe dosages after 52 days’ trial.

Parameters Substrate Fe dosage (mg/L)

0 50 200 1000 2000
Dmax (mL/gVS/day) No NaOH 52.6 ± 2.5a 58.5 ± 2.5 51.9 ± 0.6 53.9 ± 1.9 41.1 ± 1.1
4% NaOH 72.8 ± 7.4b 86.9 ± 6.5 89.4 ± 3.3 75.0 ± 3.8 68.1 ± 2.7
6% NaOH 111.6 ± 7.6c 133.5 ± 2.5 143.9 ± 3.4 136.1 ± 1.1 107.6 ± 3.0
TDmax (day) No NaOH 15a 11 10 7 3
4% NaOH 3b 3 3 3 3
6% NaOH 3c 3 3 3 3
CBy (mL/gVS) No NaOH 530.6 ± 41.2a 595.9 ± 31.7 613.5 ± 26.0 616.5 ± 31.2 574.6 ± 39.3
4% NaOH 570.7 ± 56.2b 672.6 ± 30.9 702.6 ± 31.7 618.1 ± 68.2 666.7 ± 64.9
6% NaOH 670.9 ± 66.2c 752.6 ± 66.0 837.2 ± 63.2 831.1 ± 41.9 813.9 ± 60.7
CMy (mL CH4/gVS) No NaOH 302.8 ± 23.4a 342.1 ± 17.5 335.6 ± 14.2 350.2 ± 17.6 298.2 ± 20.4
4% NaOH 313.3 ± 30.7b 372.0 ± 17.0 396.2 ± 17.8 341.4 ± 37.6 356.4 ± 34.7
6% NaOH 370.9 ± 38.0c 435.3 ± 40.0 455.6 ± 34.7 490.2 ± 25.2 464.7 ± 34.9
⁄
MCr (%) No NaOH 38–62a 50–62 50–63 45–69 45–66
4% NaOH 42–66b 42–64 52–67 45–64 45–66
6% NaOH 46–68c 50–68 50–66 51–68 44–63

Dmax: Maximum daily biogas yield. TDmax: Time of the maximum daily biogas yield.
CBy: Cumulative biogas Yield. CMy: Cumulative methane yield ⁄MCr: methane content range.
a,b,c
Represent; the control value (No NaOH and Fe dosage), values for 4% NaOH and 6% NaOH pretreatment alone, respectively.

sure of the effect of the two treatments on the gas yield. From
Fig. 2, the peak biogas production time was shorter for pretreated
samples (2 weeks) than for the untreated straw (3 weeks). The lag
phase of 7 days observed with the untreated straw can be attribu-
ted to the complex structure of the cell wall that required several
days for hydrolysis of the holocellulose by the microorganisms
[24].
Table 3 shows the different TDTs for biogas production. Com-
pared to the TDT100 of the control (48 days), NaOH pretreatment
reduced the TDT100 to 19 days and 10 days for 4% and 6% pretreat-
ment, respectively. The shortening in digestion time for pretreated
straw could be due to easier holocellulose degradation by the
microorganisms since NaOH pretreatment results in increase of
accessible surface area, decrease in degree of polymerization and
crystallinity, separation of structural linkages between lignin, cel-
lulose and hemicelluloses, and disruption of lignin structure [17].
3.4. Synergistic effect of NaOH pretreatment and Fe dosing on biogas
and methane yield
The results displayed in Fig. 2 and Table 2 show that daily and
cumulative produced biogas from combined Fe and NaOH pretreat-
ments is not only significantly higher than from untreated straw
but also higher than from either Fe dosing or NaOH pretreatments
alone. For instance, at a dosing of 200 mg/L Fe, the biogas yields in
4% and 6% NaOH pretreated straw were 702.6 mL/gVS and
837.2 mL/gVS, respectively, whereas only 570.74 mL/gVS and
670.9 mL/gVS were obtained for 4% NaOH and 6% NaOH alone.
Likewise, the biogas yield (613.5 mL/gVS) from the group with only
Fe dosing (200 mg/L) was lower than that from the group of com-
bined treatments. This trend was observed with other Fe dosages.
Additionally, the maximum daily biogas yield (Dmax) in com-
bined treatments (50–1000 mg/L Fe and NaOH pretreatment)
was higher than in treatments with NaOH only or Fe dosing alone.
The achieved Dmax values for 4% NaOH pretreated straw at different
Fe dosing were 86.9 mL/gVS for 50 mg/L Fe, 89.4 mL/gVS for
200 mg/L Fe, 75.0 mL/gVS for 1000 mg/L Fe, and 68.1 mL/gVS for
2000 mg/L Fe. The Dmax values for 6% NaOH pretreated straw were
133.5 mL/gVS for 50 mg/L Fe, 143.9 mL/gVS for 200 mg/L Fe,
136.1 mL/gVS for 1000 mg/L Fe, and 107.6 mL/gVS for 2000 mg/L
Fe (Table 2). Conversely, the Dmax values for untreated straw (no
NaOH) at different Fe dosages were 58 mL/gVS for 50 mg/L Fe,
51.9 mL/gVS for 200 mg/L Fe, 53.9 mL/gVS for 1000 mg/L Fe, and
41.1 mL/g/VS for 2000 mg/L Fe. The most optimized production
which gave the highest Dmax values and cumulative biogas yields
in both 4% and 6% NaOH pretreated straw groups resulted from
200 mg/L Fe. Surprisingly, Fe dosing alone at 200 and 1000 mg/L
gave 7.6% and 8.2% higher biogas yields than that obtained from
4% NaOH pretreatments (Table 2). However, considering the big
standard deviation in the data, statistical analysis (Kruskal–Wallis
One Way ANOVA) showed no significance difference (q = 2.880–
3.188; p > 0.05).
Fig. 3 and Table 2 give a further comparison of the energy con-
version efficiency based on achieved methane yield computed as
methane production per unit volatile solids loaded (mL/gVS). The
achieved maximum daily methane yield (Dmmax) and cumulative
methane yields from combined treatments were higher than those
obtained from untreated straw and single factor treatment cases
(Fe dosing or NaOH pretreatment alone). For instance, at a dosing
of 1000 mg/L Fe, cumulative methane yield of 6% NaOH pretreated
straw was 490.2 mL CH4/gVS that is 61.8% and 32.16% higher than
untreated (302.8 mL CH4/gVS) and 6% NaOH pretreatment alone
(370.9 mL CH4/gVS). Likewise, the Dmmax values of 6% NaOH pre-
treatment were 65.9 mL CH4/gVS for 50 mg/L Fe, 64.3 mL CH4/
gVS for 200 mg/L Fe, 77.9 mL CH4/gVS for 1000 mg/L Fe and
58.3 mL CH4/gVS for 2000 mg/L Fe all of which are higher than
either untreated or single treatment (Fig. 3).
From Fig. 3 and Table 2, it can be observed that at combined
treatment using 6% NaOH and different Fe dosages yielded cumu-
lative methane yield range from 435.3 to 490.2 mL CH4/gVS which
were 43.7–61.9% higher than untreated (302.8 mL CH4/gVS) and
17.4–32.4% higher than 6% NaOH pretreatment alone (370.9 mL
CH4/gVS). This result implies that with combined treatments it is
possible to obtain more bioenergy with the same amount of straw.
In practice this provides logistical and production advantages of
reducing the straw volume while increasing throughput.
The ANOVA results (Table 5) revealed that biogas production
from NaOH pretreated straw was significantly (p < 0.05) higher
than from untreated straw. Additionally, the cumulative biogas
yield from straw dosed with iron between 200 and 1000 mg/L
was significantly higher than from the control. However, there
was no significant difference in biogas yield when compared to
control at low (50 mg/L) and very high (2000 mg/L) Fe dosage. Fur-
thermore, there was no significant difference in the methane con-
 S. Khatri et al. / Applied Energy 158 (2015) 55–64 61

100 600
A Untreated B

Cumulave methane yield (mL/gVS)

Daily methane yield (mL/gVS-d)
500
80

400
60
300
40
200

20
100

0 0
100 600
C 4% NaOH pretreated D

Cumulave methane yield (mL/gVS)

Daily methane yield (mL/gVS-d)

500
80

400
60

300
40
200

20
100

0 0
100 600
E 6% NaOH pretreated F

Cumulave methane yield (mL/gVS)

Daily methane yield (mL/gVS-d)

500
80

400
60
300
40
200

20
100

0 0
0 5 10 15 20 25 30 35 40 45 50 55 0 5 10 15 20 25 30 35 40 45 50 55
Time (d) Time (d)

Fe 0 mg/L Fe 50 mg/L Fe 200 mg/L Fe 1000 mg/L Fe 2000 mg/L

Fig. 3. Daily methane yield and cumulative methane yield of untreated (no NaOH) (A and B), 4% NaOH pretreated (C and D) and 6% NaOH pretreated maize straw (E and F).

Table 3
Effect of treatments on technical digestion time (days) of maize straw.

Fe (mg/L) Treatment groups

No NaOH + Fe 4% NaOH + Fe 6% NaOH + Fe
T50 T80 T100 T50 T80 T100 T50 T80 T100
0 13a(265.3) 17a(424.4) 48a(530.6) 5b 9b 19b 4c 7c 10c
50 10 13 22 4 7 12 3 6 8
200 11 14 20 4 7 10 3 5 7
1000 10 14 21 5 9 16 3 5 7
2000 9 18 35 6 10 17 3 7 8

T50, T80, T100 = time taken in days to achieve 50, 80 and 100% of total cumulative biogas yield based on the control (no NaOH and Fe dosage).
() = 50%, 80% and 100% value of total cumulative biogas yield from the control in mL/gVS.
a,b,c
Represent; the control value (No NaOH and Fe dosage), value for 4% NaOH and 6% NaOH pretreatment alone, respectively.
62 S. Khatri et al. / Applied Energy 158 (2015) 55–64

Table 4 Table 5
Difference of biogas production (%) compared to control. Two way ANOVA depicting differences in cumulative biogas yield at different Fe
dosages within the no NaOH, 4% NaOH and 6% NaOH pretreatments.
Difference in cumulative biogas production (%)
Source of variation DF SS MS F P
Fe (mg/L) No NaOH + Fe 4% NaOH + Fe 6% NaOH + Fe
* b c Pretreatment 2 99685.419 49842.71 49.779 <0.001
0 530.60 7.57 26.46
Fe concentration 4 27429.542 6857.385 6.849 0.011
50 12.32 26.77 41.85
Residual 8 8010.225 1001.278
200 15.64 32.43 57.80
Total 14 135125.186 9651.799
1000 16.20 16.51 56.65
2000 8.31 25.65 53.40
Comparisona Diff of Means t P P < 0.050
b,c
Represent values for 4% NaOH and 6% NaOH pretreatment alone, respectively.
*
Represents cumulative biogas yield (mL/gVS) of the control (no NaOH and Fe Comparisons for factor: pretreatment
dosage) from which the percentage difference for the combined treatment was 6% vs. No NaOH 194.924 9.74 <0.001 Yes
calculated. 6% vs. 4% 135 6.746 <0.001 Yes
4% vs. No NaOH 59.924 2.994 0.017 Yes
Comparisons for factor: Fe concentration
tent of the produced biogas between alkaline treatments at differ- 200 vs. 0 127.033 4.917 0.012 Yes
1000 vs. 0 97.837 3.787 0.047 Yes
ent Fe dosing. This proves that neither treatment nor combination
2000 vs. 0 94.320 3.651 0.05 Yes
of treatments contributed to an increase in the methane content 50 vs. 0 82.960 3.211 0.084 No
(Table 2). 200 vs. 2000 32.713 1.266 0.748 No
In order to determine the process stability, the pH and VFA 200 vs. 1000 29.197 1.130 0.748 No
accumulation were measured. The results show that the pH of a
All pairwise multiple comparison procedures (Holm-Sidak method).
the reactors with different Fe dosages fluctuated between pH
7.17 and 7.81, the optimal range. However, at 2000 mg/L Fe, the
pH range was slightly lower in both the pretreated and untreated through increased throughput without affecting the gas produc-
straw (pH 6.71–7.33). This phenomenon could be attributed to tion volume.
the formation of iron (II) hydroxide under reductive conditions At a dosage of 2000 mg/L Fe, non-alkaline pretreated straw
due to large addition of an electron acceptor. Conversely, acetic required a longer technical digestion time (TDT100) of 35 days.
acid was the major VFA (75.7–79.5%) produced followed by butyric However, at this same Fe dosing, 4% and 6% NaOH pretreatments
acid (9.8–12.7%) and propionic acid (9.8–12.8%). Overall, the total had shorter TDT100 values of 17 days and 8 days respectively
VFA production and pH ranges in all reactors were below process (Table 3). Even at high Fe dosage (2000 mg/L Fe), shorterTDT100
inhibition levels throughout the experiment. Further details are may still be attributed to the improved access of biodegradable
included in the supplementary material (Supplementary Figs. 1 material within straw as a result of alkaline pretreatment.
and 2).
Combining NaOH pretreatment and Fe dosing (200 mg/L)
resulted in two-fold and four-fold percent increase in biogas yield 3.6. Preliminary economic feasibility evaluation and environmental
over untreated straw at same Fe dosing for 4% NaOH and 6% NaOH impacts of proposed treatments
respectively (Table 4). Previous studies on the effect of Fe dosage
on anaerobic digestion of different substrates support our findings. In order to make rough preliminary economic considerations,
For instance, Jarvis et al. [39] evaluated grass-clover silage and we compared the added value related to the increase of bio-
reported a stimulatory effect of Fe for biogas production at range methane relative to the additional costs of chemicals. During com-
of 109–132 mg/L Fe. As well, Hinken et al. [31] reported a 35% putation, the achieved methane values of the discussed batch
increase in biogas production at 205 mg/L Fe during anaerobic experiment were extrapolated to substrate requirements of
digestion of maize silage. In this study, the synergistic effect can 1000 kg maize straw (VS: 85.5%). Based on 55–60% methane con-
be explained by the increased accessibility and microbial degrada- tent in the biogas, we evaluated the energy content per m3 in terms
tion of the biodegradable fraction of the straw as a result of NaOH of kilo Watt hour (kW h) equivalent and used the per kW h energy
pretreatment and Fe dosing. price as a surrogate price for 1 m3 producible methane. The reason
for using the surrogate price is that in China the market price of
biogas (biomethane) is not standard (relatively low; 1–2 CNY per
3.5. Synergistic effect of NaOH pretreatment and Fe dosing on m3) given that biogas production runs on 80% government subsi-
technical digestion time dies [3].
The calculated values are shown in supplementary Table 2.
In the control group (no NaOH pretreatment or Fe dosing), the From this table, with exception of 6% NaOH alone, and combination
technical digestion time (TDT100) was 48 days. Compared to the of 6% NaOH and 50–200 mg/L Fe for the rest treatments the cost of
control, addition of Fe at dosages 200–1000 mg/L to untreated chemicals is much higher than the marginal values of methane
straw reactors reduced the TDT100 to 20–21 days (Table 3). Com- produced. However, it is worth noting that fully optimized condi-
bining Fe dosing between 200–1000 mg/L with 4% NaOH and 6% tions were not considered in during the lab batch process. Perhaps
NaOH pretreated straw further shortened the TDT100 to an average better industrial scale optimization with different operation modes
of 10 days and 7 days, respectively. The shortened time and conse- (for instance; continuous fed digestion) could increase methane
quently, more biogas production resulting from the combination of yield values and achieve economic feasibility especially for the
NaOH pretreatment and Fe dosing may be related to the improved range 200–1000 mg/L Fe. Moreover, further investigations need
access of degradable, volatile solids by NaOH pretreatment and in to be done in the area of process optimization using different
the presence of Fe as an enzyme co-factor contributing to acceler- anaerobic digestion modes in order to obtain data, which could
ated degradation. In practice, the shortened TDT using the same be used for process scale-up. Additionally, full scale feasibility
capacity digester means that combining both treatments would studies should be done at optimum scale production to ascertain
have a production benefit by increasing straw digestion capacity. sustainable economic advantages.
The results also demonstrate the potential plant design benefits From an ecological perspective, special care needs to be paid to
where shorter retention periods would reduce production costs reduction and/or recovery of Na+ ions in the digestate because the
 S. Khatri et al. / Applied Energy 158 (2015) 55–64
disposal of high Na+-containing slurry could lead to negative envi-
ronmental impacts such as soil salinization and water pollution
[18]. A recent study by Chen [40] showed it is possible to recover
NaOH used in biomass pretreatment during anaerobic digestion.
In his findings, Chen reports improved biogas yield from grape
pomace and rice straw attributed to recycling NaOH during the
digestion process. This process could help to reduce the pretreat-
ment cost and mitigate the potential environmental pollution
caused by waste Na+ disposal.
It is also worth noting that although Fe is considered a
macronutrient in plant nutrition, high residual Fe in the digestate
can result in its high loading in the soil a phenomenon that could
affect soil chemistry. Remarkably, it has been reported that Fe con-
centration in the range of 500 mg/L and above can cause toxicity in
the soil [41,42]. In larger scale operations, the optimal Fe dosing for
optimal biomethane yield achievement with low residual Fe in the
digestate is an area of interest for future research. Chemical precip-
itation [43] and other electrochemical recovery mechanisms such
electro-kinetics and electro-depositing [44] if integrated with post
digestate treatment a reasonable amount of Fe can be retained and
recycled back into biomass pretreatment and macronutrient sup-
ply in the digester and thus reducing the threat of its accumulation
in the soil when the residual digestate slurries are applied as bio-
fertilizer. To this end, future research could focus on the effect of
residual Na+ at various NaOH pretreatment and residual Fe from
various dosing on the soil microbial activities and soil properties
after the application of digestate slurry.
4. Conclusions
 Alkaline pretreatment increased the methane yield by 3.5% (4%
NaOH) and 22.5% (6% NaOH) and shortened the technical diges-
tion time (TDT) from 48 days to 19 days and 10 days in 4% and
6% NaOH pretreated straw respectively.
 Fe dosing (200–1000 mg/L) alone stimulates biogas yield incre-
ment(percent) to more than half of that obtained from 6% NaOH
but two fold higher than 4% NaOH pretreatments. This indicates
that Fe dosing could be used in place of NaOH pretreatment, but
the TDT will be 10 days longer.
 Combining NaOH pretreatment and Fe dosing resulted in a syn-
ergistic effect achieving 21.8% and 56.2% higher cumulative
methane yield than from the control and shortened TDT from
48 days to 13 days and 7 days in 4% NaOH and 6% NaOH pre-
treatment respectively. The synergistic effect may be attributed
to the improved accessibility of the biodegradable fraction on
the straw due to alkaline treatment while Fe increased the
microbial enzyme activity and consequently, higher biogas
yield.
 There was no significant difference in the methane content of
biogas produced from combined Fe and NaOH compared to Fe
or NaOH treatments alone.
Overall, from our findings, we suggest that combining NaOH
pretreatment to achieve lignocellulose breakdown and FeCl2 as
the source of iron micronutrient for microorganisms could over-
come the digestibility barrier and provide novelty anaerobic diges-
tion approach for optimizing processes fed with lignocellulosic
materials to produce energy. However, further investigations using
different digestion operation modes would be required in order to
obtain data, which could be used for process scale-up.
Acknowledgements
This work was supported by grants from Beijing Municipal
Science and Technology Projects (Nos. D141100001214001 and
63
D141100001214002). We are grateful to Lu Qimin and BahHami-
dou for their valuable assistance in the laboratory. We likewise
greatly appreciate the critical and constructive comments from
the anonymous reviewers, which have helped improve this
manuscript.
Appendix A. Supplementary material
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.apenergy.2015.
08.045.
References
[1] Chen Y, Hu W, Feng Y, Sweeney S. Status and prospects of rural biogas
development in China. Renew Sustain Energy Rev 2014;39:679–85.
[2] Cheng S, Li Z, Mang H-P, Huba E-M. A review of prefabricated biogas digesters
in China. Renew Sustain Energy Rev 2013;28:738–48.
[3] Chen Y, Yang G, Sweeney S, Feng Y. Household biogas use in rural China: a
study of opportunities and constraints. Renew Sustain Energy Rev
2010;14:545–9.
[4] Chandra R, Takeuchi H, Hasegawa T. Methane production from lignocellulosic
agricultural crop wastes: a review in context to second generation of biofuel
production. Renew Sustain Energy Rev 2012;16:1462–76.
[5] Weiland P. Biogas production: current state and perspectives. Appl Microbiol
Biotechnol 2010;85:849–60.
[6] Ward AJ, Hobbs PJ, Holliman PJ, Jones DL. Optimisation of the anaerobic
digestion of agricultural resources. Bioresour Technol 2008;99:7928–40.
[7] Li J, Wei L, Duan Q, Hu G, Zhang G. Semi-continuous anaerobic co-digestion of
dairy manure with three crop residues for biogas production. Bioresour
Technol 2014;156:307–13.
[8] Chen M, Zhao J, Xia L. Enzymatic hydrolysis of maize straw polysaccharides for
the production of reducing sugars. Carbohydr Polym 2008;71:411–5.
[9] Bah H, Zhang W, Wu S, Qi D, Kizito S, Dong R. Evaluation of batch anaerobic co-
digestion of palm pressed fiber and cattle manure under mesophilic
conditions. Waste Manage 2014;34:1984–91.
[10] Li Y, Zhang R, Chen C, Liu G, He Y, Liu X. Biogas production from co-digestion of
corn stover and chicken manure under anaerobic wet, hemi-solid, and solid
state conditions. Bioresour Technol 2013;149:406–12.
[11] Zhou S, Zhang Y, Dong Y. Pretreatment for biogas production by anaerobic
fermentation of mixed corn stover and cow dung. Energy 2012;46:644–8.
[12] Niu M, Pang X, Chen S. The study of influencing factors to corn straw mixed
with pig effluent anaerobic fermentation. Proc Environ Sci 2011;8:54–60.
[13] Xi Y, Chang Z, Ye X, Xu R, Du J, Chen G. Methane production from wheat straw
with anaerobic sludge by heme supplementation. Bioresour Technol
2014;172:91–6.
[14] Zhou A, Guo Z, Yang C, Kong F, Liu W, Wang A. Volatile fatty acids productivity
by anaerobic co-digesting waste activated sludge and corn straw: effect of
feedstock proportion. J Biotechnol 2013;168:234–9.
[15] Ahring BK, Biswas R, Ahamed A, Teller PJ, Uellendahl H. Making lignin
accessible for anaerobic digestion by wet-explosion pretreatment. Bioresour
Technol 2015;175:182–8.
[16] Behera S, Arora R, Nandhagopal N, Kumar S. Importance of chemical
pretreatment for bioconversion of lignocellulosic biomass. Renew Sustain
Energy Rev 2014;36:91–106.
[17] Hendriks ATWM, Zeeman G. Pretreatments to enhance the digestibility of
lignocellulosic biomass. Bioresour Technol 2009;100:10–8.
[18] Zheng Y, Zhao J, Xu F, Li Y. Pretreatment of lignocellulosic biomass for
enhanced biogas production. Prog Energy Combust Sci 2014;42:35–53.
[19] Chen X, Zhang Y, Gu Y, Liu Z, Shen Z, Chu H, et al. Enhancing methane
production from rice straw by extrusion pretreatment. Appl Energy
2014;122:34–41.
[20] Xiao X, Zhang R, He Y, Li Y, Feng L, Chen C, et al. Influence of particle size and
alkaline pretreatment on the anaerobic digestion of corn stover. BioResources
2013;8:5850–60.
[21] Liu S, Wu S, Pang C, Li W, Dong R. Microbial pretreatment of corn stovers by
solid-state cultivation of Phanerochaete chrysosporium for biogas production.
Appl Biochem Biotechnol 2014;172:1365–76.
[22] Chandra R, Takeuchi H, Hasegawa T, Kumar R. Improving biodegradability and
biogas production of wheat straw substrates using sodium hydroxide and
hydrothermal pretreatments. Energy 2012;43:273–82.
[23] Sapci Z. The effect of microwave pretreatment on biogas production from
agricultural straws. Bioresour Technol 2013;128:487–94.
[24] Zheng M, Li X, Li L, Yang X, He Y. Enhancing anaerobic biogasification of corn
stover through wet state NaOH pretreatment. Bioresour Technol
2009;100:5140–5.
[25] Zhu J, Wan C, Li Y. Enhanced solid-state anaerobic digestion of corn stover by
alkaline pretreatment. Bioresour Technol 2010;101:7523–8.
[26] Demirel B, Scherer P. Trace element requirements of agricultural biogas
digesters during biological conversion of renewable biomass to methane.
Biomass Bioenergy 2011;35:992–8.
64 S. Khatri et al. / Applied Energy 158 (2015) 55–64
[27] Sawatdeenarunat C, Surendra KC, Takara D, Oechsner H, Kumar Khanal S.
Anaerobic digestion of lignocellulosic biomass: challenges and opportunities.
Bioresour Technol 2014. http://dx.doi.org/10.1016/j.biortech.2014.09.103.
[28] Schattauer A, Abdoun E, Weiland P, Plöchl M, Heiermann M. Abundance of
trace elements in demonstration biogas plants. Biosyst Eng 2011;108:57–65.
[29] Ortner M, Rachbauer L, Somitsch W, Fuchs W. Can bioavailability of trace
nutrients be measured in anaerobic digestion? Appl Energy 2014;126:190–8.
[30] Chen Y, Cheng JJ, Creamer KS. Inhibition of anaerobic digestion process: a
review. Bioresour Technol 2008;99:4044–64.
[31] Hinken L, Urban I, Haun E, Urban I, Weichgrebe D, Rosenwinkel KH. The
valuation of malnutrition in the mono-digestion of maize silage by
anaerobic batch tests. Water Sci Technol: J Int Assoc Water Pollut Res
2008;58:1453–9.
[32] Takashima M, Shimada K, Speece RE. Minimum requirements for trace metals
(iron, nickel, cobalt, and zinc) in thermophilic and mesophilic methane
fermentation from glucose. Water Environ Res: Res Publ Water Environ Fed
2011;83:339–46.
[33] Zandvoort MH, van Hullebusch ED, Fermoso FG, Lens PNL. Trace metals in
anaerobic granular sludge reactors: bioavailability and dosing strategies. Eng
Life Sci 2006;6:293–301.
[34] Oleszkiewicz JA, Sharma VK. Stimulation and inhibition of anaerobic processes
by heavy metals—a review. Biol Wastes 1990;31:45–67.
[35] Song Z, Gaihe Yang, Liu X, Yan Z, Yuan Y, Liao Y. Comparison of seven chemical
pretreatments of corn straw for improving methane yield by anaerobic
digestion. PLoS ONE 2014;9:e93801.
View publication stats
[36] Zhang W, Lang Q, Wu S, Li W, Bah H, Dong R. Anaerobic digestion
characteristics of pig manures depending on various growth stages and
initial substrate concentrations in a scaled pig farm in Southern China.
Bioresour Technol 2014;156:63–9.
[37] APHA. Standard methods for the examination of water and wastewater. 21st
ed. Washington, DC: American Public Health Association; 2005.
[38] Scharer JM, Moo-young M. Methane generation by anaerobic digestion of
cellulose-containing wastes. Advances in biochemical engineering, vol.
11. Berlin, Heidelberg: Springer; 1979. p. 85–101.
[39] Jarvis Å, Nordberg Å, Jarlsvik T, Mathisen B, Svensson BH. Improvement of a
grass-clover silage-fed biogas process by the addition of cobalt. Biomass
Bioenergy 1997;12:453–60.
[40] Chen X. Development of effective pretreatment and bioconversion systems for
converting organic residuals to bioenergy. University of California, Davis 2012;
PhD Dissertation; publication number 3555276 [328p].
[41] Sahrawat KL. Iron toxicity in wetland rice and the role of other nutrients. J
Plant Nutr 2005;27:1471–504.
[42] Becker M, Asch F. Iron toxicity in rice—conditions and management concepts. J
Plant Nutr Soil Sci 2005;168:558–73.
[43] Zhu N-M, Qiang L, Guo X-J, Hui Z, Yu D. Sequential extraction of anaerobic
digestate sludge for the determination of partitioning of heavy metals.
Ecotoxicol Environ Saf 2014;102:18–24.
[44] Hanay O, Hasar H, Kocer NN. Effect of EDTA as washing solution on removing
of heavy metals from sewage sludge by electrokinetic. J Hazard Mater
2009;169:703–10.