Renewable Energy 115 (2018) 76e86

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Renewable Energy
journal homepage: www.elsevier.com/locate/renene

Biogas production from co-digestion Salvinia molesta and rice straw

and kinetics
Iqbal Syaichurrozi
Department of Chemical Engineering, University of Sultan Ageng Tirtayasa, Postal Code 42435, Cilegon, Indonesia

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

Article history: The biogas fermentation co-digestion Salvinia molesta (SM) and rice straw (RS) was investigated within a
Received 13 June 2017 wide range of SM:RS of 100:0e0:100 (mass ratio). This study used batch anaerobic digesters in
Received in revised form laboratory-scale operated at mesophilic temperature in 30 days. The optimum SM:RS ratio was 60:40
23 July 2017
e0:100 (C/N of 29.50e39.17) producing total biogas yield of 103.83e113.92 mL/g VS. The highest biogas
Accepted 11 August 2017
yield (113.92 6.90 mL/g VS) was obtained from SM:RS of 40:60 (C/N of 34.80). The biogas produced
Available online 12 August 2017
from SM:RS of 40:60 contained 60.58% CH4, 38.69% CO2, 0.73% H2. Fitting error between measured and
predicted total biogas yield for 30 days fermentation by using modied Gompertz, Cone, First Order
Keywords:
Biogas
model was 0.96e6.45%, 0.14e3.52%, and 1.97e15.25% respectively. Furthermore, Cone model was used to
Co-digestion design the anaerobic batch digester volume and develop the kinetic model of VS degradability rate. The
C/N ratio t correlation of measured and predicted VS removal was R2 0.992.
Kinetic model 2017 Elsevier Ltd. All rights reserved.
Rice straw
Salvinia molesta

1. Introduction
Salvinia molesta (SM) is a free-oating aquatic weed that is one
of the fastest growing aquatic plants in the world. The growth rate
of this plant is faster than that of water hyacinth [1]. The water
hyacinth has doubling time of 7e12 days, whereas the SM has
doubling time of 3e10 days [2]. Because of its high growth rate and
doubling time, SM can cover the entire surface of water on rice
elds in short time in Indonesia. The presence of SM in water bodies
of rice elds will reduce the efciency of irrigation systems and
reduce the effectiveness of fertilizer for rice plants, so that the total
production of rice plants will decrease. Hence, SM is one of the
most problems for Indonesian farmers in growing rice plants.
At the moment in Indonesia, the farmers just drain SM to the
rivers, or pile and burn SM, or spray the chemical agent to hamper
the SM growth. However, those methods are not effective and
efcient, also make the new problems. SM drained into the rivers
will grow quickly and cover the river surface, so the rivers are losing
their water through evapotranspiration. Besides that, SM will
disturb the movement of aquatic organisms including shes and
decrease the amount of oxygen so that the aquatic organisms will
be death. Burning SM and using chemical agent is not eco-friendly.
E-mail address: iqbalsyaichurrozi@gmail.com.
Hence, the new method must be applied to control the SM growth.
The best solution is using anaerobic digestion (AD). With this
method, SM will be converted into biogas and fertilizer. The biogas
can be used to produce electricity that is useful for rural commu-
nities. Also, the biogas can be used in greenhouse farming [3]. By
using a greenhouse concept, all plant growth factors can be
controlled and maintained, so that it can produce greenhouse
products maximally. However, it need a very expensive cost espe-
cially heating cost that has almost 60e80% of total production cost.
Therefore, the biogas can substitute the heating energy need. Some
authors reported that the SM is potential as biogas feedstock [1,2,4].
Indonesian SM contains high carbon to nitrogen ratio (C/N), which
is 57.39 (Table 1). One of the most important parameters in AD is C/
N ratio. During fermentation, degradation of carbon-rich substrates
will generate Volatile Fatty Acids (VFAs) in large amount that causes
dropping pH sharply, so that it disturbs the bacterial activities.
Meanwhile, degradation of nitrogen-rich substrates will generate
ammonium (NH 4 eN) or ammonia (NH3-N) easily. Abundant of
these in substrates can hamper the bacterial growth. Anaerobic
bacteria need carbon to produce biogas and nitrogen to build their
cell structure [5,6]. Anaerobic bacteria especially methanogenic
bacteria thrive in the substrates containing C and N in good ratio.
Therefore, the nitrogen source must be added to adjust the C/N of
SM substrate.
The co-digestion methods is better than addition of synthetic

http://dx.doi.org/10.1016/j.renene.2017.08.023
0960-1481/ 2017 Elsevier Ltd. All rights reserved.
 I. Syaichurrozi / Renewable Energy 115 (2018) 76e86 77

Table 1 of volatile solid (VS). This model could predict the degradation rate
Characteristics of SM and RS. of VS during fermentation and calculate the VS removal in the end
Component SM RS of fermentation. This model was original and has not been found by
Total solid (TS) (%) 86.98 94.48
other authors yet.
Ash (%) 36.17 20.75
Volatile solid (VS) (%) 50.81 73.73 2. Methods
Crude ber (%) 22.09 35.90
Crude carbohydrate (%) 46.93 64.17
Crude protein (%) 3.07 8.68 2.1. Wastewater and inoculums
Crude lipid (%) 0.81 0.87
Lignin (%) 11.73 7.72 SM and RS were obtained from rice elds in Bayah Regency,
Hemicellulose (%) 2.78 6.14 Banten Province, Indonesia. Then, these biomasses were analyzed
Cellulose (%) 1.83 9.00
NDF (%) 16.34 25.71
through proximate and van soest methods to obtain the data of
ADF (%) 13.56 16.71 their chemical compositions (Table 1). Meanwhile, the total organic
Total Organic Carbon (TOC) (%) 28.23 40.96 carbon (TOC) and total nitrogen (TN) were calculated using relation,
Total Nitrogen (TN) (%) 0.49 1.39 TOC VS/1.8 [9] and TN protein/6.25 [10]. The C/N value was
C/N 57.39 29.50
obtained from TOC/TN. The rumen uid was used as inoculums. In
Remarks: SM, Salvinia molesta; RS, Rice straw; VS TS - Ash; NDF, Neutral detergent this study, rumen uid in fresh condition was obtained from cow
ber; ADF, Acid detergent ber; C/N, Carbon/Nitrogen ratio; TN Crude protein/ slaughterhouse in Serang City, Banten Province, Indonesia. Rumen
6.25; TOC VS/1.8; C/N TOC/TN.
uid contained Clostridium sp., Clostridium sporogenes, Clostridium
butyricum and rich methanogenic bacteria.
nitrogen source (such as urea), because it can treat two wastes in
the same time and it can decrease the operational cost which is 2.2. Experimental set up
buying synthetic nitrogen. In addition, co-digestion methods can
enhance the biogas volume [7,8]. Rice straw (RS) is the potential co- Anaerobic digesters were made from polyethylene bottles hav-
digestion partner of SM. RS in Indonesia contains low C/N (below ing volume of 600 mL. The bottles were plugged with rubber plug
30) that is obtained from rice eld where SM lives. Hence, the and equipped with valve for biogas measurement. Biogas formed
transportation cost is cheap. Every single rice plant will produce RS was measured by liquid displacement method as also has been
as much as 58% from its total mass. used by the other authors [11e15]. In this method, each digester
Based on that, this study investigated the anaerobic co-digestion was connected to gas collector that was reserved cylindrical glass.
of SM and RS. The utilization of RS as nitrogen source to SM has not The connection was done using connecting tube. Each gas collector
been reported by other authors yet. Previous studies just investi- was immersed in through of water to ensure complete sealing.
gate the mono-digestion SM. Mathew et al. [2] studied biogas Biogas formed from digesters was collected by the downward
production from SM at mesophilic temperature and cow dung was displacement of water.
used as inoculum. Total biogas formed was 221 L/kg VS. That value
was less than total biogas from water hyacinth (552 L/kg VS). 2.3. Experimental design
O'Sullivan et al. [4] reported that mono-digestion of SM at meso-
philic temperature produced less total biogas (155 L/kg VS) than Anaerobic digesters of experimental laboratory using 600-mL
water hyacinth (267 L/kg VS) and cabomba (221 L/kg VS). Abbasi volumes were operated in batch system. Total mass of SM and RS
and Nipaney [1] produced biogas from SM (total biogas of 6.7 L/kg) mixing of 10 gr was put into the digesters. Water was added with
with ratio of SM:water of 1:7 (w/w). substrate:water ratio of 1:7 %w/w. This ratio was adapted from
The aim of this study was to investigate the effect of SM:RS in study of Abbasi and Nipaney [1]. Rumen uid as methanogenic
mass ratio of 100:0, 80:20, 60:40, 40:60, 20:80, 0:100 to biogas bacteria provider was added into the digester as much as 25 mL
production on anaerobic digestion performance in batch test. The [13]. Substrates were varied at SM:RS of 100:0, 80:20, 60:40, 40:60,
SM:RS ratio affected the C/N ratio in substrates. In this study, the 20:80, 0:100. Furthermore, initial pH for all variables was adjusted
author used laboratory-scale batch anaerobic digesters (poly- 7.0 0.1 by using NaOH solution 1 N. The variable in this work can
ethylene bottles, volume 600 mL) operated at mesophilic temper- be seen in Table 2.
ature (~30  C) and at pressure of 1 atm in 30 days. The experimental
data was used to make kinetic model of biogas production using 2.4. Experimental procedures
some models. We compared the modied Gompertz model, rst
order kinetic model and cone model in predicting biogas yield and Fermentation was done for 30 days at mesophilic temperature
nd which the model is the best of these models. Furthermore, the (~30  C) and at pressure of 1 atm. Hence, the obtained experimental
best model was used to develop the kinetic model of degradability data can be used to make kinetic model of biogas production well.

Table 2
Variation of SM:RS ratios.

VW:TW Substrate (gram) Water (mL) Rumen VS (gr) Total Organic Total Nitrogen C/N Fiber contents
uid (mL) Carbon (gr) (gr)
SW (gr) RS (gr) Hemicellulose (%VS) Cellulose (%VS) Lignin (%VS) NDS (%VS)

100:0 10 0 70 25 5.08 2.82 0.05 57.39 5.47 3.60 23.09 67.84

80:20 8 2 70 25 5.54 3.08 0.07 45.85 6.23 5.89 19.73 67.12
60:40 6 4 70 25 6.00 3.33 0.09 39.17 6.87 7.83 16.88 66.50
40:60 4 6 70 25 6.46 3.59 0.10 34.83 7.42 9.50 14.40 66.00
20:80 2 8 70 25 6.91 3.84 0.12 31.77 7.90 10.94 12.32 65.52
0:100 0 10 70 25 7.37 4.10 0.14 29.50 8.32 12.20 10.47 65.13

Remarks: SW, Salvinia molesta; RS, Rice straw; VS, Volatile solid; C/N, Carbon/nitrogen ratio.
78 I. Syaichurrozi / Renewable Energy 115 (2018) 76e86

Other authors also conducted anaerobic digestion test for 30 days
to make kinetic model of biogas production [11,12]. Biogas formed
was measured once in two days to know biogas production by
using water displacement method. The pH of substrate in the di-
gesters was measured by using pH meter once in two days. First, the
digesters were opened. The rubber plug, which plugged the di-
gesters, was taken. Then, substrates were taken from the digesters
as much as 10 mL. After that, the digesters were plugged quickly.
The pH level of substrates taken was measured by using pH meter.
This procedure was done very quickly to keep the comfortable
condition for methanogenic bacteria. This pH measurement was
also applied by Syaichurrozi et al. [5]. Furthermore, in the end of
fermentation, substrates were analyzed to know the nal TS (total
solid) and ammonium concentration (NH 4 eN) according to Stan-
dard Methods APHA 22nd edition 2012 [16]. Furthermore, VS was
calculated by VS TS e Ash [17]. The VS removal was calculated
using Equation (1) proposed by Di Maria et al. [18]. The ammonia
concentration (NH3-N) and volatile fatty acids (VFAs) concentration
was calculated using Equation (2) proposed by El-Mashad et al. [19]
and (3) proposed by Paul and Beauchamp [20] respectively. The
methane concentration of methane in biogas produced from each
variable was measured using GC-TCD (Gas Chromatography-
Thermal Conductivity Detector) Shimadzu 8A. Biogas was
collected in gas chamber during 30 days. Then, the cumulative
biogas in gas chamber was taken as much as 20 mL for analyzing
methane using GC-TCD.
initial VS  final VS
VS Removal%  100%
initial VS
" #1

 10pH
NH3  N NH  N  1 ;
100:1075 T
4 2725
T absolute temperature; K
U the maximum biogas production rate (mL/g VS. day)
l lag phase period or minimum time to produce biogas (days)
t cumulative time for biogas production (days)
e mathematical constant (2.718282)
khyd hydrolysis rate constant (/day)
n shape factor
k the biogas rate constant (/day)
3. Results and discussions
3.1. Biogas production
The daily and cumulative biogas yield (mL/g VS) during the
digestion of different SM:RS ratio were shown in Fig. 1. The biogas
yield increased until end of fermentation (30th day) for all variables
(Fig. 1(b)). The peak value of daily biogas productions was obtained
to be 1.97 0.00, 5.69 3.34, 13.25 2.25, 13.39 1.16, 14.32 0.43,
11.73 1.56 mL/g VS after four, fourteen, sixteen, eighteen, eigh-
teen, eighteen days of digestion from SM:RS ratio of 100:0, 80:20,
60:40, 40:60, 20:80, 0:100 respectively (Fig. 1(a)). The total biogas
yield was 6.30 0.00, 32.76 18.32, 107.54 18.51, 113.92 6.89,
104.93 0.22, 103.83 7.53 for 100:0, 80:20, 60:40, 40:60, 20:80,
0:100 variables respectively (Fig. 1(b)).
The combination of SM:RS ratio affected the difference of C/N in
the substrates. The C/N ratios in the substrates of 100:0, 80:20,
60:40, 40:60, 20:80, 0:100 were 57.39, 45.85, 39.17, 34.80, 31.77,
29.50 respectively (Table 2). Generally, the optimum C/N ratio was
(1) in range of 25e30 [23]. In this research, the more the RS was added,
(2)

VFAs
pH 9:43  2:02 h
 i; R2 0:955
NH3  N NH
4  N
(3)

2.5. Kinetic model of biogas production

Biogas production kinetic was modeled through modied

Gompertz model [5], Cone model [21], First order kinetic model
[22]. Kinetic of biogas production in batch condition was assumed
that had correspondence to specic growth rate of methanogenic
bacteria in digesters. Kinetic constant of ym, l, U, khyd, n, k was
determined by using non-linear regression with help of polymath
software. The equations of modied Gompertz model (4), cone
model (5), rst order kinetic model (6) were shown below:
  
U:e
yt ym:exp  exp l  t 1 ; t  0 (4)
ym

ym
yt
n ; t > 0 (5)
1 khyd $t

yt ym1  expk$t; t  0 (6)

where:

y(t) the cumulative biogas at digestion time t days (mL/g VS) Fig. 1. The Effect of SM:RS Ratio on (a) Daily Biogas, (b) Cumulative Biogas, (c) pH
ym the biogas production potential (mL/g VS) Prole.

the less the C/N ratio of the substrates. Total biogas yield increased
with decreasing C/N from 57.39 (SM:RS 100:0) until 34.80
(SM:RS 40:60). Furthermore, biogas decreased at C/N below
34.80 (C/N of 31.77e29.50). The variable of 100:0e60:40 had too
high C/N ratio (contained too low nitrogen source), so that anaer-
obic bacteria could not build their cell structure and nally could
not grow well. Meanwhile, variable of 20:80e0:100 contained C/N
ratio that was included in optimum range proposed by Kayhanian
and Hardy [23]. However, the two variables generated the less total
biogas yield than the variable 40:60. The optimum C/N ratio in
anaerobic digestion depended on the substrates used. This study
used SM and RS that contained high lignocelluloses, especially
lignin (Table 1).
According to Triolo et al. [17], the fractions of lignocellulosic
bers are hemicellulose, cellulose, lignin, NDS. The composition of
these fractions in each substrate (100:0e0:100) in this study was
shown in Table 2. In anaerobic digestion, cellulose and hemicel-
lulose are fermentable after hydrolysis which makes lignocellu-
losic biomass a suitable feedstock for biogas production. However,
the presence of lignin makes lignocellulosic biomass resistant to
degradation by microbes [24]. Lignin is a natural complex polymer
and is constructed of phenylpropane units (such as coniferyl
alcohol and sinapyl alcohol) with hydroxyl, methoxyl, and
carbonyl functional groups [25]. Lignin has a role for the cross-
linking between cellulose and hemicellulose to form a rigid
three-dimensional structure of the cell wall providing mechanical
strength. Because of its rigid structure, lignocellulose is very
slowly bioconvertible in anaerobic digestion. Lignin is non-
degradable and suppresses the degradation of hemicellulose and
celluloses [17,24].
From Table 2, the more RS portion in the mixture substrate of
SM and RS, the less the lignin content in the mixture. Whereas, the
cellulose and hemicellulose content increased with the increasing
of RS portion in the mixture. According to Zheng et al. [24],
decreased lignin content leads to increase biogas yield and biode-
gradability rate of lignocellulosic biomass. In this study, decreasing
of lignin from 23.09%VS (SM:RS 100:0; C/N 57.39) until 14.40%
VS (SM:RS 40:60; C/N 34.83) could increase the total biogas
yield from 6.30 0.00 until 113.92 6.90 mL/g VS. Furthermore,
lignin content below 14.40%VS (C/N below 34.83) decreased total
biogas yield. Therefore, C/N and lignin content in substrates had
140
120
Total Biogas Yield (mL/g VS)
100
80
60
40
20
0
C/N=57.39; C/N=45.85; C/N=39.17; C/N=34.80; C/N=31.77; C/N=29.50;
Lignin
23.09%VS 19.73%VS 16.88%VS 14.40%VS 12.32%VS 10.47%VS
Fig. 2. Relationship between lignin concentration, C/N ratio and total biogas yield.
I. Syaichurrozi / Renewable Energy 115 (2018) 76e86 79
important correlation to convert lignocellulosic biomass to biogas
successfully.
From Fig. 2, C/N ratio of 45.85e57.39 resulted biogas yield of
6.30 0.00e32.76 18.32 mL/g VS (below 50 mL/g VS). Meanwhile,
C/N ratio of 29.50e39.17 resulted biogas yield of
103.83 7.53e113.92 6.90 mL/g VS (more than 100 mL/g VS).
Ryckebosch et al. [26] stated that the more concentration of CH4 in
biogas, the better the quality of biogas. The common range of CH4
concentration was 40e75% [26]. From Table 3, C/N of 29.50e39.17
resulted biogas with high CH4 concentration which was
49.05e74.34%. Whereas, at C/N of 45.85e57.39, biogas which was
produced contained CH4 of 23.18e29.09% (below 40%). Therefore,
the optimum range of C/N to produce with high quantity (high total
biogas yield) and quality (high CH4 concentration) from lignocel-
lulosic substrates was 29.50e39.17. Furthermore, for the better
results, the C/N of 29.50e34.80 could produce biogas with high CH4
concentration (60.58e74.34%).
Another study showed the same results with this study. Zheng
et al. [27] found that the C/N optimum to produce biogas from co-
digestion of dairy manure and switchgrass was 29.40e32.60 that
was obtained from manure:switchgrass ratio of 2:2e1:3. In that
range, methane yield produced was 142.5e155.1 mL/g VS. Whereas,
substrates having C/N out of the optimum range (less than 29.40
and more than 32.60) produced methane yield of 88.9e133.7 mL/g
VS. Like SM and RS, dairy manure and switchgrass also contained
high lignin, with the value of 7.8 and 14.5 %TS respectively.
Therefore, the results of this study emphasized that the optimum
range of C/N for lignocellulosic substrates was approximately
30e40.
_
Baltrenas and Misevicius [28] reported that if there are more
carbohydrates in the substrate, the concentration of methane can
reach 50e60%. However, if there is more fat or protein, the con-
centration of methane can reach 75e90%. That was same with this
study. The lower the C/N value of substrates, the higher the CH4
concentration in biogas (Table 3). It means that the more nitrogen
(protein) portion in the substrates, the higher the CH4 concentra-
tion in biogas. The C/N value from 57.39 to 29.50, CH4 concentration
increased from 29.09 to 74.34%.
In this study, the co-digestion of SM:RS produced more total
biogas yield than mono-digestion SM. The most total biogas yield of
113.92 6.89 mL/g VS (methane 60.58%) was obtained from co-
Lignin Lignin Lignin Lignin Lignin
80 I. Syaichurrozi / Renewable Energy 115 (2018) 76e86

Table 3
The results of anaerobic digestion.

SM:RS C/N Initial pH Final pH NH

4 -N (mg/L) NH3-N Ratio of NH
4 : NH3 TAN (mg/L) Total VFAs Total Biogas Biogas Composition
(mg/L) (mg/L) (mL/g VS)
CH4 (%) CO2 (%) H2 (%)

100:0 57.39 7.0 0.1 6.8 0.2 134.55 0.63 99.5:0.5 135.18 178.00 6.30 0.00 29.09 70.76 0.15
80:20 45.85 7.0 0.1 6.6 0.1 nd nd nd nd nd 32.76 18.32 23.18 74.10 2.72
60:40 39.17 7.0 0.1 6.7 0.1 317.70 1.35 99.6:0.4 319.05 426.45 107.54 18.51 49.05 50.87 0.08
40:60 34.80 7.0 0.1 6.7 0.1 329.85 1.31 99.6:0.4 331.16 447.55 113.92 6.90 60.58 38.69 0.73
20:80 31.77 7.0 0.1 6.6 0.3 366.75 1.15 99.7:0.3 367.90 515.43 104.93 0.22 74.27 25.71 0.02
0:100 29.50 7.0 0.1 6.5 0.3 491.40 1.23 99.8:0.2 492.63 714.56 103.83 7.53 74.34 25.61 0.04

Remarks: SM, Salvinia molesta; RS, Rice straw; C/N, Carbon/Nitrogen ratio; TAN, Total Ammonium Nitrogen; VFAs, Volatile Fatty Acids; TAN NH
4 NH3; nd, not analyzed;
CH4, Methane; CO2, Carbon Dioxide; H2, Hidrogen.

digestion SM:RS of 40:60. This result was more than total biogas
yield from mono-digestion of SM in study of Abbasi and Nipaney
[1], which was 6.7 mL/g TS (methane 40e78%). In other hand, the
study of O'Sullivan et al. [4] and Mathew et al. [2] produced more
total biogas yield than this study. They produced biogas from
mono-digestion SM with biogas yield 155 mL/g VS (methane 50%)
and 221 mL/g VS (methane up to 62%) respectively. The difference
of these results might be caused by the difference of chemical
compositions of SM used, especially the C/N ratio and lignin con-
tent. The SM used in this study might be contained more lignin
content than that in the other studies.
Co-digestion had no signicant effect on substrate pH. The
initial pH for all variables was 7.0 considered suitable for bacteria
involved in anaerobic digestion. The pH prole during fermentation
for 30 days ranged between 6.5 and 6.9 that was shown in Fig. 1(c).
Whereas, the nal substrate pH was 6.5e6.8. The pH uctuated
throughout the digestion period, indicating occasional decrease
and increase, because of VFAs and ammonium/ammonia produc-
tion from degradation of carbon and nitrogen contents.
3.2. Ammonium, ammonia, VFAs
During fermentation, nitrogen source in the substrates was
decomposed to be ammonium (NH 4 ) or ammonia (NH3). Ammo-
nium or ammonia was used by microbe to build their cell structure.
Composition of ammonium and ammonia was depended on sub-
strate pH. Ammonium ions and ammonia have permanent equi-
librium: NH4 4 NH3 H and NH 
4 OH 4 NH3 H2O [5,29].
The more acid of substrate pH, the more ratio of ammonium:
ammonia. At substrate pH of 9.0, the ratio of ammonium: ammonia
is 70:30 and at substrate pH of 7.0, the ratio of ammonium:
ammonia is 99:1 [30]. Moreover, at substrate pH > 9.25, ammonia is
full dominant in the substrate [31] and pH < 7.0, ammonium ions is
full dominant in the substrate [5]. In this study, nal pH for all
variables was 6.5e6.8 (<7.0), so that the ammonium: ammonia
ratio was in range of 99.5:0.5-99.8-0.2 (Table 3). That proved that
ammonium ion was dominant in the system.
The nal ammonium in this study was 134.55e491.40 mg/L,
whereas the nal ammonia was 0.63e1.35 mg/L. Total ammonia
nitrogen (TAN) was obtained from NH 4 NH3 with the value of
135.18e492.63 mg/L. The more nitrogen (the less C/N ratio) in the
substrate, the more ammonium was resulted (Table 3). Correlation
between C/N and TAN was shown in Fig. 3, with equation of
y 10.88x 748.4 (R2 0.90), where y was TAN concentration
(mg/L) and x was C/N value. Rajagopal et al. [32] stated that TAN
concentration of 50e200 mg/L is benecial for bacterial growth.
That means, in the range of concentration, bacteria can thrive in the
substrate. Variable SM:RS of 100:0 (C/N 57.39) had TAN con-
centration of 135.18 mg/L (Table 3). Furthermore, TAN concentra-
tion of 200e1000 mg/L was no antagonistic effect [32]. Whereas,
TAN concentration of 1500e10,000 mg/L was start inhibition and
that of 30,000 was toxic for bacteria [29]. The other variables,
SM:RS of 60:40e0:100 (C/N 29.50e39.17), resulted TAN of
319.05e492.63 mg/L (Table 3). These concentrations were not
negative effect on bacterial activity, because these were below
1000 mg/L. It can be concluded that TAN generated from all vari-
ables (100:0e0:100) in this study had not negative effect. However,
variable of 100:0 had less biogas yield (6.30 mL/g VS) than variable
of 60:40e0:100 (C/N 29.50e39.17). That phenomenon showed
that variable of 100:0 contained too high C/N ratio and lignin, so

600
TAN concentration (mg/L)

500

400

300

200 y = -10.88x + 748.4

R = 0.90
100

0
0 10 20 30 40 50 60 70
C/N

Fig. 3. Correlation between C/N and TAN concentration.

 I. Syaichurrozi / Renewable Energy 115 (2018) 76e86 81

that anaerobic bacteria could not grow well and produced biogas in
little amount.
Volatile fatty acids (VFAs) were generated during decomposition
of carbon source. The accumulation of VFAs concentration can
decrease substrate pH sharply. The low pH can disturb the stability
of anaerobic digestion that depends upon the maintenance of a
delicate biochemical balance between the acidogenic and meth-
anogenic microorganisms [32]. Methanogenic microorganisms
were more sensitive with low pH than acidogenic microorganisms.
In this study, substrate pH during fermentation was 6.5e6.9
(Fig. 1(c)). Whereas, the nal substrate pH was 6.5e6.8 (Table 3).
These values were still in neutral pH range. The presence of VFAs
could not drop the pH, because its concentration was not excess.
The VFAs for all variables was 178.00e714.56 mg/L (Table 3). Ac-
cording to Table 3, the more accumulation of TAN, the more accu-
mulation of VFAs. Niu et al. [33] also reported that increasing of TAN
concentration from 5000 mg/L to 15,000 mg/L, the VFAs accumu-
lation increased sharply from 2000 to 15,000 mg/L.
3.3. Kinetic model
The kinetic parameters of ym, l, U, khyd, n, k in modied Gom-
pertz, Cone and First order model were estimated based on the best
t of the studied models and the results were summarized in
Table 4. By plotting experimental and simulation data, we got the
graph as shown in Fig. 4.
3.3.1. Using modied Gompertz model
From Table 4, substrate of SM:RS ratio of 60:40e0:100 (C/
N 29.50e39.17) had more value of ym than variable of
100:0e80:20 (C/N 45.85e57.39). That means ratio of
60:40e0:100 generated the maximum biogas yield in larger
amount (120.68e140.49 mL/g VS) compared to ratios of
100:0e80:20 (6.05e33.35 mL/g VS). That was due to anaerobic
bacteria in the comfortable metabolism conditions supported by
the mixing feedstock of SM:RS of 60:40e0:100. The more value of
ym, the more value of m. Kinetic constant of m was maximum biogas
production rate, so that the more biogas production rate, the more
total biogas formed [5]. Substrate of 60:40e0:100 had the higher
value of m (4.80e5.86 mL/g VS. d) than substrate of 100:0e80:20
(0.35e2.17 mL/g VS. d). The good ratio of SM:RS was 60:40e0:100
which had C/N of 29.50e39.17 and the best C/N was 34.80 obtained
from SM:RS of 40:60.
The value of l indicated the time that was required for anaerobic
bacteria to adapt in the substrates before produced biogas [13].
Therefore, the variable having low value of l, needed just little time
to produce biogas. Based on that, bacteria in substrate with SM:RS
ratio of 100:0 needed less time to adapt which was 0.14 days.
Whereas, bacteria in substrate of 80:20e0:100 needed the longer
time than variable of 100:0 (SM alone). Substrate containing high
proteins needed the longer time than substrate containing high
carbohydrate [13]. The high carbohydrate was contained in SM
alone (SM:RS of 100:0, C/N of 57.39) so that bacteria took short time
to degrade that (0.14 days). Moreover, the more presence of RS (C/N
of 29.50) in substrate, the more time needed by anaerobic bacteria
to convert organic content into biogas (SM:RS of 80:20e0:100; l of
5.03e5.83 days).
3.3.2. Cone model
The predicted maximum biogas yield (ym) of variable of SM:RS
of 60:40e0:100 (132.95e183.99 mL/g VS) was larger than that of
variable SM:RS of 100:0e80:20 (8.04e35.13 mL/g VS). The biggest
value of ym was in variable of SM:RS of 40:60 (C/N 34.80). The
khyd showed the hydrolysis rate of organic materials. The higher the
khyd, the easier the organic materials were degraded. From Table 4,
the less value of l obtained from modied Gompertz, the more
value of khyd. Variable of SM:RS of 100:0 had lowest value of l (0.14
days) and highest value of khyd (0.08/day). SM contained high C/N
ratio (57.39) and RS contained low C/N ratio (29.50). The time
needed by bacteria to degrade carbohydrate was less than to
degrade protein. The more RS presented in the mixture, the less C/N
ratio (the higher nitrogen content). Hence, bacteria needed longer

Table 4
Results from using modied Gompertz, Cone, rst order kinetic model.

SM:RS Ratio

100:0 80:20 60:40 40:60 20:80 0:100

Modied Gompertz Model

l (days) 0.14 5.69 5.03 5.83 5.65 5.15
m (mL/g VS.d) 0.35 2.17 5.39 5.38 5.86 4.80
R2 0.959 0.995 0.998 0.998 0.995 0.997
ym (mL/g VS) 6.05 33.35 127.16 140.49 120.68 137.01
Predicted biogas yield (mL/g VS)-30 d 5.89 32.14 109.17 112.83 108.24 106.74
Measured biogas yield (mL/g VS)-30 d 6.30 32.76 107.54 113.92 104.93 103.83
Difference between measured and predicted biogas yield (%) 6.45 1.90 1.51 0.96 3.16 2.81

Cone Model
khyd (/day) 0.08 0.07 0.05 0.04 0.06 0.05
n 1.31 3.14 2.18 2.12 2.66 2.14
R2 0.966 0.995 0.997 0.998 0.992 0.993
ym (mL/g VS) 8.04 35.13 155.53 183.99 132.95 157.92
Predicted biogas yield (mL/g VS)-30 d 6.10 32.31 110.12 113.76 108.62 103.53
Measured biogas yield (mL/g VS)-30 d 6.30 32.76 107.54 113.92 104.93 103.83
Difference between measured and predicted biogas yield (%) 3.07 1.39 2.40 0.14 3.52 0.29

First-Order Kinetic Model

k (/day) 0.06 0.03 0.03 0.03 0.03 0.03
R2 0.972 0.929 0.919 0.900 0.911 0.915
ym (mL/g VS) 7.31 55.82 156.38 163.12 161.61 156.85
Predicted biogas yield (mL/g VS)-30 d 6.17 32.10 98.36 96.55 98.83 92.22
Measured biogas yield (mL/g VS)-30 d 6.30 32.76 107.54 113.92 104.93 103.83
Difference between measured and predicted biogas yield (%) 1.97 2.02 8.54 15.25 5.81 11.18

Remarks: SM, Salvinia molesta; RS, Rice straw; ym, the biogas production potential; m, the maximum biogas production rate; l, lag phase period or minimum time to produce
biogas; khyd, hydrolysis rate constant; n, shape factor; k, the biogas rate constant; R2, correlation coefcient.
82 I. Syaichurrozi / Renewable Energy 115 (2018) 76e86
Fig. 4. Comparison of Experimental Data and Simulation Data using Modied Gom-
pertz, Cone and First Order Kinetic Model at Various SM:RS Ratios.
time to adapt, and hydrolysis phase was carried out in long time.
The khyd value in SM:RS of 100:0e80:20 (0.07e0.08/day) was more
than that in SM:RS of 60:40e0:100 (0.04e0.06/day).
3.3.3. Using rst order kinetic model
Variable of SM:RS of 60:40e0:100 had ym value of
156.38e163.12 mL/g VS. Meanwhile SM:RS of 100:0e80:20 had ym
value of 7.31e55.82 mL/g VS. Hence, the mixture of SM:RS in range
of 60:40e0:100 (C/N 29.50e39.17) was good mixture substrate
for bacterial activity. Kae et al. [34] reported that, the more the
value of k, the faster the rate of biogas generation. From Table 4,
substrate of 100:0 (SM alone) had the most k value (0.06/day) of all
variables. Meanwhile substrate of SM:RS of 80:20e0:100 had the
less k value (0.03/day). Raposo et al. [35] stated that degradation of
substrate containing high protein needed more time than that of
substrate containing high carbohydrates. RS contained high nitro-
gen and low carbohydrate content (C/N below 30), so that anaer-
obic bacteria were need a long time to degrade organic materials in
RS. The value of k in rst order kinetic model had good correlation
with l value in modied Gompertz and with khyd value in Cone
model. The more value of k, the less value of l and the more value of
khyd.
Although SM:RS of 100:0 had the most k value, it resulted the
least biogas yield. That might be caused by the high lignin content.
The high carbon content made it easily produced biogas in rst
time, but the production of biogas was low because lignin was not
easy to be degraded. Therefore, the total biogas yield in prediction
was just 7.31 mL/g VS.
3.3.4. Comparison the modied Gompertz, cone, and rst order
kinetic model
The difference between the measured biogas and predicted
biogas for 30 days observed in modied Gompertz model was
0.96e6.45%, in Cone model was 0.14e3.52%, in First order kinetic
model was 1.97e15.25% (Table 4). Clearly, Cone model and modied
Gompertz model tted the actual evolution of biogas production
successfully because both of them had tting error below 10%. That
was also strongly supported by their high correlation coefcient (R2
of 0.966e0.998 for Cone model and R2 of 0.959e0.998 for modied
Gompertz model).
The rst order kinetic model gave the satisfactory result with
error of 10% or less only if the lag period needed by bacteria was
very short [35]. Kae et al. [34] reported that degradation of high
protein substrate needed a long lag time and high carbohydrate
substrate needed a short time. In this study, at substrate of SM:RS of
100:0, the lag time was the shorter (l was 0.14 days) than the other
substrates, so that the tting error at rst order kinetic was little,
which was 1.97%. The SM contained high proportion of carbohy-
drates (C/N 57.39). Meanwhile, with the more RS addition into
SM substrate (80:20e0:100), the lag time (l) was longer (5.03e5.83
days), so that the tting error at rst order kinetic became bigger,
which was up to 15.25%. With that value, rst order kinetic model
was not allowed to be used in predicting biogas production from
co-digestion SM and RS, because there were some variables having
the tting error more than 10%. Whereas, modied Gompertz
model gave the satisfactory result in predicting biogas production
for all variables (tting error 0.96e6.45%). This study similar with
study of Zhang et al. [36]. Zhang et al. [36] stated that rst order
kinetic gave the bad prediction in anaerobic digestion of pig
manure and dewatered sewage sludge with the tting error of
1.5e14.2%. In other hand, modied Gompertz could give better
prediction with error of 0.0e3.7% (Table 5).
Meanwhile, some authors found the different results with this
study. Budiyono et al. [22] and Zhen et al. [21] reported that both
rst order kinetic and modied Gompertz model gave the good
tting (error <10%) (Table 5). Budiyono et al. [22] predicted biogas
yield from vinasse waste containing high carbohydrate. Thus,
biogas was generated at the rst time of vinasse fermentation (l of
0e2.24 days). Furthermore, Zhen et al. [21] also found that methane
yield from co-digestion of waste activated sludge (WAS) and Egeria
densa (E.d.) could be predicted using both rst order kinetic (tting
error of 4.0e7.1%) and modied Gompertz model (tting error of
4.4e7.3%). The WAS and E.d. contained high carbohydrate and low
protein contents.
Generally, Cone model and modied Gompertz model could be
used to predict biogas production from co-digestion SM and RS
because both of them had tting error was below 10% (Table 4).
Moreover, the prediction using Cone model was better than that
using modied Gompertz. The tting error of Cone model was
below 4% (error 0.14e3.52%), whereas the tting error of modied
Gompertz model was below 7% (error 0.96e6.45%). Other authors
also reported the same results. Syaichurrozi et al. [13] reported that
Cone model (error 0.193e2.809%) give the better prediction than
modied Gompertz (error 0.316e3.225%) (Table 5). Zhen et al. [21]
also reported that Cone model had error of 1.0e2.8% and modied
Gompertz had error of 4.4e7.4% (Table 5). Cone equation contained
the shape factor n, so that it had exibility in modeling various
pattern of cumulative biogas production from experiment.
This study also showed that Cone model had a better t to the
experiment data than rst order kinetic model. This result was in
line with study of Zhen et al. [21]. Fitting error of Cone model and
rst order kinetic model was 1.0e2.8% and 4.0e7.1% respectively.
However, in contrast to this result, Li et al. [37] reported that Cone
model could not predict the biogas yield from dairy manure well
(Table 5). According to Li et al. [37], Cone model successfully
simulate biogas production from chicken manure and rabbit
manure with error below 10%. Furthermore, in predicting biogas
yield from dairy manure, it had error up to 13.50% (Table 5). That
might be caused by the characteristics of dairy manure, so that
biogas was very easy to be produced in the rst day, but after a few
fermentation days, biogas production very low. This biogas
 I. Syaichurrozi / Renewable Energy 115 (2018) 76e86 83

Table 5
Comparison between this result and other results in modied Gompertz, rst order kinetic and Cone model to predict biogas yield.

Substrate Difference between measured and predicted biogas (%) References

Modied Gompertz First Order Kinetic Cone Model

Tofu wastewater 0.316e3.254 e 0.193e2.809 [13]

Co-digestion of waste activated sludge and Egeria dense 4.4e7.3 4.0e7.1 1.0e2.8 [21]
Dairy manure e 0.70e3.05 5.62e13.50 [37]
Chicken manure e 2.77e70.88 0.54e9.62 [37]
Rabbit manure e 0.22e3.06 6.74e8.98 [37]
Brewery grain waste 3.2 19.5 e [34]
Bread waste 1.6 9.2 e [34]
Pacic saury sh waste 0.7 13.6 e [34]
Mackerel sh waste 6.1 29.8 e [34]
Cuttle sh waste 13.7 37.1 e [34]
Vinasse 0.76e3.14 1.54e7.5 e [22]
Co-digestion of pig manure and dewatered sewage sludge 0.0e3.7 1.5e14.2 e [36]
Co-digestion vinasse and tofu-processing wastewater 1.18e9.79 0.82e17.33 e [6]
Co-digestion of Salvinia molesta and rice straw 0.96e6.45 1.97e15.25 0.14e3.52 This study

production pattern was more suitable with rst order kinetic model Vgc (mL) y(t)-(mL/g VS)  m (mass of VS fed into the
(error 0.70e3.05%) than Cone model (error 5.62e13.50%). Mean- digester) (9)
while, the biogas production in this study was almost stable until
the end fermentation, so Cone model was better than rst order Cone model was used in this design because it had least tting
kinetic. error and best R2. The equation of Cone model was shown in
Equation (5a)

3.4. VS removal ym
yt
n (5a)
1 khyd $t
Volatile solid (VS) in the substrates were converted to biogas in
anaerobic digestion. The bacterial activity consisted of four steps i.e. Substituting Equation (9) into (8) to get (10)
hydrolysis, acidogenesis, acetogenesis and methanogenesis.
Amount of VS value that was degraded by bacteria was called VS Vdigester 3  yt  m (10)
removal. Based on Table 6, the more the formation of biogas yield,
the more the VS was removed to biogas. The prediction model of VS Substituting Equation (10) into (5a) to get (11)
degradability rate will be discussed in next discussion point. The
ym
value of VS removal for all variables can be seen in Table 6. The Vdigester 3 
n  m (11)
relationship between total biogas yield formed and VS removal can 1 khyd $t
be obtained by plotting the value of total biogas yield (mL/g VS)
against the value of VS removal (%) to get straight line equation Equation (11) was used to design volume of batch anaerobic di-
y 5.102  e 174.7, where y total biogas yield formed (mL/g VS) gesters. Kinetic constant of ym and khyd value can be seen in Table 4.
and x VS removal (%). Value of m was initial VS that can be seen in Table 6. Retention time
t for all variables was 30 days. Hence, volume of digesters for
variable of SM:RS of 100:0, 80:20, 60:40, 40:60, 20:80, 0:100 was
3.5. Design of anaerobic batch digester using cone model 111.26, 633.92, 2309.22, 2542.54, 2580.81, 2599.64 mL respectively.

In anaerobic batch digester, ratio of gas chamber volume-: 3.6. Prediction of VS degradability rate
anaerobic digester volume was 1:3 [15]. Gas chamber (Vgc) was
proportional to the volume of biogas formed. In this study, this ratio In this study, authors made prediction about VS degradability
was used to design the batch digester. rate through Cone model. The correlation between VS degradability
and biogas yield at any time can be developed with assuming all VS
Vgc 1/3$Vdigester (7) contents are converted into biogas, as shown in Fig. 5.
From Fig. 5, can be deduced that:
3 Vgc Vdigester (8)
VSo  VSt yt
(12)
VSo ym

Table 6 
VSo  VSt
Comparison between measured and predicted VS removal. yt  ym (13)
VSo
SM:RS Initial VS (g) Measured VS Predicted VS

Final (g) Removal (%) Final (g) Removal (%) where, VSo was total VS that can be removed; VSt is VS at any time;
100:0 6.08 3.58 41.09 3.578 43.08
yt is biogas yield at any time; ym is biogas yield maximum.
80:20 6.54 3.09 52.74 3.090 52.82 Cone model equation:
60:40 6.99 3.40 51.36 3.400 51.60
40:60 7.45 2.91 61.01 2.909 61.17 ym
yt
n (5b)
20:80 7.92 4.02 49.26 4.019 49.34
0:100 8.37 4.32 48.36 4.319 48.59
1 khyd $t

Remarks: SM, Salvinia molesta; RS, Rice straw; VS, Volatile Solid. Substituting Equation (5b) into (13) to get (14)
84 I. Syaichurrozi / Renewable Energy 115 (2018) 76e86

VSo
ym - yt ym

VSo - VSt

Biogas yield
VS

yt
VSt

t Time (t)
Fig. 5. VS transformation into biogas during anaerobic degradation.

10
(a)
9
VS content in the substrates (gram)

8
7
SM:RS=100:0
6
SM:RS=80:20
5
SM:RS=60:40
4 SM:RS=40:60
3 SM:RS=20:80
2 SM:RS=0:100
1
0
0 5 10 15 20 25 30
Time (day)

61
(b)
Predicted VS Removal (%)

56
y = 0.9164x + 4.6989
R = 0.9925
51

46

41
41 46 51 56 61
Measured VS Removal (%)

Fig. 6. Application of VS degradability kinetic model in (a) prediction of degradation rate of VS during fermentation, (b) comparison of measured and prediction of VS removal.
 I. Syaichurrozi / Renewable Energy 115 (2018) 76e86
 aquatic weeds of santiniketan through anaerobic digestion, Clean. Technol.
VSo  VSt ym
 ym
n (14)
VSo 1 k $t
hyd

VSo  VSt 1 (Cabomba caroliniana) and Salvinia (Salvinia molesta), J. Ecol. Eng. 36 (2010)

n (15)
VSo 1 khyd $t
1
VSt VSo 
n  VSo (16)
1 khyd $t
In the end of fermentation process, substrate still contained
amount of VS called nal VS (VSf), so Equation (16) become (17)
1
VSt VSo 
n  VSo VSf (17)
1 khyd $t
where, VSt was VS at any time (g); VSo was total VS that can be
removed (g) (VSo VSi e VSf, VSi was initial VS (g), VSf was nal VS
(g)); khyd was hydrolysis rate constant (/day); n was shape factor; t
was cumulative time for biogas production (days).
Equation (17) can be used to get prediction of VS degradability
during fermentation process in the digesters. The curve kinetic
model of VS degradability for all variables can be seen in Fig. 6(a).
The predicted nal VS and VS Removal were shown in Table 6.
Furthermore, the accuracy between measured VS removal (%) ob-
tained from experiment test and predicted VS removal (%) obtained
from the kinetic model of Equation (17) was shown in Fig. 6(b).
4. Conclusion
Variables of SM:RS of 100:0, 80:20, 60:40, 40:60, 20:80, 0:100
generated total biogas of 6.30 0.00, 32.76 18.32, 107.54 18.51,
113.92 6.90, 104.93 0.22, 103.83 7.53 mL/g VS respectively. The
more portion of RS in the substrates, the less the C/N of substrates.
Furthermore, the less the C/N of substrates, the higher the TAN and
total VFAs concentration in the systems. The optimum of C/N was
29.50e39.17 (SM:RS of 60:40e0:100). In that range, biogas con-
tained CH4 concentration of 49.05e74.34%. The rst order model
was not suitable for predicting biogas production because it had
tting error more than 10% (1.97e15.25%). Meanwhile, the modi-
ed Gompertz and Cone model had tting error below 10%
(0.96e6.45% and 0.14e3.52% respectively). The Cone model was
chosen to design volume of batch anaerobic digester and develop
kinetic model of VS degradability during fermentation. The high
lignin content in the substrates might cause total biogas yield in
this study that was less than total biogas yield in the other studies.
Therefore, author recommended studying pre-treatment of SM to
increase biogas yield.
Acknowledgements
The author thanks to LPPM Untirta and Ministry of Research,
Technology and Higher Education for nancial support. Also, the
author thanks to Mr. Suhirman and Mr. Topik Hidayat about their
help in collecting data in the conversion energy laboratory.
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