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Abstract
A comprehensive study has been made on biogas kinetics from municipal waste. A 10 dm3 anaerobic batch digester equipped with a
mechanical agitator has been used under controlled environment (pH 6.8, temperature = 40 C) for this purpose. Influent slurry concentration
varied from 20 to 56 kg/m3 . At 20 kg/m3 the concentration of carbohydrate, protein and fat varied from 0.83 to 2.0 kg/m3 , 0.25 to 0.70 kg/m3
and 0.26 to 0.56 kg/m3 , respectively. The biogas generated from the digester had the methane concentration as high as 80% (v/v) at an influent
slurry concentration of 20 kg/m3 on the 15th operating day of the digester. The calorific value of biogas obtained from the digester is in
the range of 27.9831.46 MJ/(N m3 ). An attempt has been made to separate both acidogenic and methanogenic bacterial consortia. It has
been noted that classical Monod-type substrate uninhibited model equation can be used to predict the growth kinetics of separated bacterial
consortia of both types. Biogas generation kinetics was studied using initial influent slurry, carbohydrate, protein and fat concentrations as
parameters. By judiciously coupling differential mass balance equations, mathematical model equations were obtained.
2005 Elsevier Inc. All rights reserved.
Keywords: Kinetic studies; Biogas; Municipal waste
1. Introduction
Efficient disposal of municipal market waste (both vegetables and non vegetables) is always a sensitive issue to civic
authorities since the presently available disposal processes
like sanitary landfill, incineration, pyrolysis, etc., are always
associated with pollution hazards posing a serious threat to
public health [1]. This problem demands necessity of a green
technology for disposal of such municipal wastes, which
should be cost effective in one hand and pollution free on the
other. Anaerobic biological treatment of market wastes may
probably be one of the promising routes which can satisfy
the above two criteria [24]. Successful application of such
anaerobic digestion technology is, however, critically dependent on proper understanding of the degradation kinetics,
judicious selection of mode of operation- batch, semi-batch
continuous, etc., and optimization of process parameter [5].
Corresponding author. Tel.: +91 33 2414 6378; fax: +91 33 2414 6378.
E-mail address: ftbe bon@yahoo.com (R. Chowdhury).
0141-0229/$ see front matter 2005 Elsevier Inc. All rights reserved.
doi:10.1016/j.enzmictec.2005.07.004
494
2.2. Methods
Nomenclature
c
d
Ks
M
Qc
r
Sl
S
Y
concentration (kg/m3 )
days
saturation constant (kg/m3 )
molecular weight
cumulative concentration
reaction rate
slurry concentration
solubility
yield coefficientsexpressed as gram
material substrate/product
consumed/produced per gram of
biomass formed
Greek letters
max
maximum specific growth rate (d1 )
Subscripts
IVII reaction numbers IVII
Bio
of biogas
carb
of carbohydrate
CH4
of methane
fat
of fat
i
concerned with any reaction i
LCFA of long chain fatty acid
pr
of protein
s
of substrate
VFA
of volatile fatty acid
x
of biomass
2.2.1. Analytical
2.2.1.1. Determination of bacterial mass concentration.
The concentration of bacterial mass in the reaction broth was
determined by dry weight method [11]. The culture broth was
centrifuged at 10,000 rpm for 20 min. The cells were washed
twice with distilled water and were taken in aluminium cups.
Aluminium cups containing the cell mass were left for 24 h
at 80 C for drying. Finally, the weight of cell mass was
measured using a sensitive Mettler-AE 240 (Switzerland) balance.
2.2.1.2. Proximate analysis of solid waste and whey. The
total solid (TS), moisture content and the volatile matter (VM)
of vegetable waste, oil cake and whey were determined by
standard methods [12].
Two grams of the sample was taken in a silica crucible and
was dried in an air oven to constant weight at 105110 C for
5 h. The weight of the residue was reported as total solid (TS)
content of the material. The difference between the weight
of sample and the residue gave the moisture content. Volatile
matter and the ash content of the waste were determined using
a muffle furnace maintained at 925 C and at 775 25 C,
respectively. Covered and open petridishes were used to hold
the samples in the furnace for the determination of volatile
matter and ash content, respectively. Volatile matter and ash
content were determined by taking the differences between
weight of the samples taken and the residues left in respective
experiments.
2.2.1.3. Determination of protein, carbohydrate and fat.
Protein, carbohydrate and fat content of vegetable waste,
whey and oil cake were determined by standard methods
[12,13].
Protein. A portion (containing approximately 0.03
0.04 g N) of the sample was weighed and transferred to a
Kjeldahl digestion flask. Eight grams of the catalyst mixture
(90% anhydrous sodium sulphate, 3.5%copper sulphate and
0.5% of selenium dioxide) and 20 ml of concentrated sulfuric
acid were added. The flask was gently heated in an inclined
position. When the initial frothing had ceased, the top of the
flask was fitted with a loose pear shaped stopper and was
heated more strongly to ensure that the liquid boiled at a
moderate rate. The flask was shaken from time to time and
the heating was continued for another one hour till the liquid became clear. Then the flask was cooled and was washed
with 400 ml ammonia free water in a distilling flask containing granulated zinc. To the receiving flask 50 ml of boric acid
solution and methyl red indicator were added. The distillation
apparatus was then connected with the delivery tube dipped in
boric acid solution. The diluted digest was made alkaline with
50% sodium hydroxide solution. After 300 ml had distilled
over, the distillate was titrated with 0.05 M sulfuric acid. The
%N2 in the sample (1 ml 0.05 M sulfuric acid 0.0014 g N)
was calculated. The crude protein figure was calculated
495
Determination of Biogas composition. The product biogas composition was analysed by gas chromatograph
(CHEMITO Model No. 8510) using thermal conductivity
detector (CHEMITO Model No. 8510) with a Porapaq-q column (Netel Chromatographs length 2 m, mesh size range
80/100). Hydrogen was used as a carrier gas at a flow rate
of 20 ml/min. The oven, injector and detector temperatures
were maintained at 60, 80 and 80 C, respectively.
2.2.1.5. Determination of caloric value of biogas. Calorific
value of the gas was determined using Junkers Calorimeter.
2.2.2. Experimental methods
2.2.2.1. Preparation of inuent digester feed. Vegetable
market waste, consisting of potato, spinach, carrot,
cauliflower, cabbage, etc., having seasonal variation, sweet
meat whey and mustard oil cake were collected from local
municipal market. The characteristics of different wastes,
determined following different analytical techniques, have
been given in Table 1. All the components were sterilized
in an autoclave at a temperature and pressure of 121 C and
0.2 MPa to ensure absence of harmful pathogens. Sterile components were subsequently taken in different proportion and
were mixed thoroughly using electrically operated mixer.
Slurry concentration was maintained at different levels using
variable quantities of tap water. Concentrations of carbohydrate, protein and fat were adjusted at desired values using
their pure sources like sucrose, casein and hydrogenated oil,
respectively.
2.2.2.2. Operation of the digester. A 10 dm3 semi-batch
biodigester having a height to diameter ratio of three, fitted with a mechanical agitator, was employed in the present
investigation. Outside the reactor a jacket was provided
through which water was circulated to ensure isothermal condition at 40 C inside the reactor. The pH of the reaction broth
was always maintained at 6.8. There was a provision for gas
outlet on the top closure from which the generated biogas
was collected continuously.
Initially the digester was fed with influent slurry to fill 80%
of the reactor volume. The volume of the seed culture added
was 10% of the reactant volume. A continuous agitation at
50 rpm was maintained during digestion process. Although
the reaction is an anaerobic one, stirring was provided only to
Table 1
Composition of different constituents of slurry
Constituents of
slurry
Moisture (%)
Total solid
(TS) (%)
Volatile solid
(VS) (%)
Protein (%)
Fat (%)
Vegetable waste
Oil cake
Whey
89.14 0.5
8.50 0.5
93.5 0.5
10.76 0.5
91.5 0.5
6.5 0.5
9.78 0.5
81.20 0.5
5.85 0.5
2.42 0.5
30.8 0.5
0.7995 0.5
0.28 0.5
9.3 0.5
0.0494 0.5
( m
)2
Carbohydrate
degradable (%)
7.18 0.5
5.005 0.5
Carbohydrate
non-degradable (%)
Ash (%)
41.0 0.5
0.98 0.5
10.30 0.5
9.2 0.5
496
maintain uniformity in the reaction broth. The slurry concentration was varied in the range of 2056 kg/m3 . According
to the constituents of vegetable waste the concentration of
soluble carbohydrate, protein and fat varied from 0.83 to
2.0 kg/m3 , 0.5 to 0.7 kg/m3 and 0.46 to 0.56 kg/m3 when the
slurry concentration was varied from 20 to 56 kg/m3 , taking
the contribution of sweet meat whey and mustard oil cake also
into account. At a slurry concentration of 20 kg/m3 , the effects
of concentration of fat, protein and carbohydrate were studied by varying their concentration from 0.26 to 0.56 kg/m3 ,
0.25 to 0.70 kg/m3 , 0.83 to 3.0 kg/m3 . Biogas coming out of
the digester was collected continuously in a sampling bottle
by the downward displacement of water and was analysed by
a gas chromatograph.
2.2.2.3. Preparation of general bacterial medium. The general bacterial medium was used for growth, enrichment and
maintenance of bacterial cultures. The composition of the
medium was as follows:
Component
Amount to be taken
Peptone (g)
Beef extract (g)
Distilled water (ml)
5.0
5.0
1000
497
culture of acidogenic bacteria maintained in general bacterial medium. To replicate the real digester condition, same
slurry having 20 kg/m3 concentration was used as the main
specific medium. Growth kinetics of acidogenic consortia
with respect to any one of the limiting substrate, namely
carbohydrate, fat and protein was determined by varying
their concentrations in the range of 0.833.0, 0.250.70,
0.260.56 kg/m3 , respectively. The concentrations of carbohydrate, fat and protein were adjusted by using their pure
3. Theoretical analysis
Since the degradation process occurs following a highly
complex route, the reaction scheme for simulation and modeling purpose can be described in the following manner:
ri =
(1)
498
Differential mass balance equations for different compounds involved in the reaction scheme mentioned earlier have been solved numerically using fourth-order
RungeKutta method.
Computation of cumulative methane and cumulative biogas production rates per unit volume of the reactor have been
done using the following expressions;
cCH4 SCH4 T
273 22.4
MCH4
Qc,CH4 =
(2)
ti
Table 2
Kinetic parameters determined using batch type (Erlenmeyer flasks) experimental data
Reaction type
Carbohydrate acidogenesis
Amino acid acidogenesis
Fat acidogenesis
VFA methanogenesis
LCFA methanogenesis
Y (yield coefficients)
Kinetic parameters
max
Ks
carb
5.17
6.40
0.550
2.440
0.559
0.518
0.20
0.10
0.049
0.019
13.15
pr
fat
LCFA
14.49
181.81
VFA
CO2
CH4
9.559
12.210
2.412
1.733
1.26
11.128
6.289
9.522
1.905
184.0
9.80
20.83
18.205
Percent standard deviation: 0.1% in all cases. All the experiments were repeated thrice. max : maximum specific growth rate (d1 ), Ks : saturation constant
(kg/m3 ), Y: absolute value of yield co-oeficient (defined in nomenclature). Culture conditions (described in text).
499
Fig. 2. Simulated (lines) and experimental time histories of volume % of methane in biogas and cumulative methane generation rate (Qc,CH4 ) in biogas with
slurry concentration (SL) as a parameter. Lines () volumetric methane percent (CH4 %); arrows ( ) represent CH4 % as ordinate and time in days as abscissa;
() cumulative methane generation rate (Qc,CH4 ) in (dm3 /(dm3 d)); arrows ( ) represent Qc,CH4 in (dm3 /(dm3 d)) as ordinate and time in days as abscissa.
Points: () 20 kg/m3 ; () 40 kg/m3 ; () 56 kg/m3 .
4.1.3.1. Slurry concentration. Fig. 5 shows that the cumulative biogas production rate increases with the increase of
slurry concentration.
Fig. 3. Simulated (lines) and experimental time histories of calorific value of biogas with slurry concentration (SL) as a parameter. Points: () 20 kg/m3 ; ()
40 kg/m3 ; () 56 kg/m3 .
500
Fig. 4. Simulated (lines) and experimental time histories of volume % of methane in biogas and cumulative methane generation rate (Qc,CH4 ) in biogas with
fat concentration in the slurry as a parameter. Lines: () volumetric methane percent (CH4 %); arrows ( ) represent CH4 % as ordinate and time in days as
abscissa; () cumulative methane generation rate (Qc,CH4 ) in (dm3 /(dm3 d)); arrows ( ) represent Qc,CH4 in (dm3 /(dm3 d)) as ordinate and time in days as
abscissa. Points signify concentration of fat: () 0.26 kg/m3 ; () 0.46 kg/m3 ; () 0.56 kg/m3 .
4.2. Discussion
4.2.1. Methane concentration, methane production rate
and caloric value of biogas
4.2.1.1. Effect of slurry concentration. Methane concentration of biogas was determined experimentally using slurry
concentration as a parameter (Fig. 2). The mathematical
model equations, namely the differential mass balance equations for various components, have been used to simulate the
volumetric concentration of methane in biogas. The simulated values of time history of methane concentration have
been represented by lines in the plots shown in Fig. 2. The
pattern of the time history curve of methane in biogas needs
to be explained. The first segment has a linear slope and it
continues up to 3 days of operation. The second segment
which continues up to 10th day after the end of the time
period of the first segment has slope lower than the first
segment. The third segment which starts from the 10th day
Fig. 5. Simulated (lines) and experimental points time histories of cumulative biogas production rate Qc,Bio in dm3 /(dm3 d)) with slurry concentration (SL) as
a parameter. Points: () 20 kg/m3 ; () 40 kg/m3 ; () 56 kg/m3 .
501
Fig. 6. Simulated (lines) and experimental points time histories of cumulative biogas production rate (Qc,Bio in dm3 /(dm3 d)) with fat concentration in the
slurry as a parameter. Points signify ratio of carbohydrate:protein:fat: () 3.2:1.92:1; () 3.2:1.92:1.76;() 3.2:1.92:2.15.
502
5. Conclusion
From the simulated and experimental results it is clear that
the variations in the initial concentrations of slurry, carbohydrate, protein and fat play a significant role regarding the
performance of anaerobic digester based on feed consisting
of municipal wastes. Present study reveals that methane concentration and the calorific value of biogas decrease with the
increase in the concentration of slurry, carbohydrate and protein, while the variation in fat concentration has the opposite
effect. A mathematical model based on the kinetic parameters
obtained from the growth kinetics of separated acidogenic
and methanogenic bacterial consortia has been developed.
Sufficient agreement between the theoretical and experimental values indicates that the model should be suitable for
similar bio-reaction systems. However, refinement of the
model is possible by the incorporation of exact enzymatic
reaction kinetics by undergoing further studies. More experimental work in this area is needed for the expansion of the
applicability of the model.
Acknowledgements
Authors highly acknowledge the financial support rendered by University Grants Commission of India to carry out
the Research and Development Project on Biogas Generation
from Food/Vegetable Wastes.
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