BIOGAS ENGINEERING

PRACTICAL COURSE WS-2015/2016

Hamburg University of Applied Sciences (HAW)
FERMENTATION BATCH TEST RUN# 248

Submitted by:
Iqbal Meskinzada Matriculation number: 2267075
Indrajeet Desai Matriculation number: 2269333

Supervisors:
Prof. Dr. Paul Scherer and Mrs. Monika Unbehauen
Table of Contents

Acknowledgement
1. Introduction and Objectives .3
2. Biogas ..................................3
3. Experimental Methodology ..4
3.1. Experimental Methodology ...4
3.2. Batches 6
4. Results ...9
4.1. Evaluation of the Measured Gas amounts .10
4.2. Calculations 17
4.2.1. I/S Ratio 17
4.2.2. Gas Quality (Methane Concentration) ...17
4.2.3. Degradation Rate of Cellulose ...18
4.3. Methane Production ..19
5. Conclusions 23

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) i

Acknowledgement: We would like to express our profound gratitude to Prof. Dr. Paul Scherer in
Biotechnology Engineering and Mrs Monika Unbehauen at Hamburg University of Applied Sciences
(HAW) Hamburg, Germany, for their supervision, guidance, constructive comments and valuable
suggestions throughout the experiment processes and discussion meetings. We would also like to express
my sincerest thanks to our colleagues who participated in the course for their huge contribution and
assistance for completing this experiment.

Introduction and Objectives: Fermentation tests are carried out in order to find out the potential of four
different substrates for biogas production. In the first place, the purpose of this lab experiment is to
analyse the suitability and gas yields of four different substrates samples namely wheat straw powder,
Grass Juice Silage, cellulose and Blanc over an incubation period of 35 days. Each of the samples is
analysed separately in details and the results are presented and compared in the coming pages. In
addition, this experiment could also be counted as a laboratory scale analysis of a large biogas power
plant. Therefore, it would give us a general idea about how a large-scale biogas power plant will operate
and we can also estimate its output parameters. In this report the following parameters will be determined

Total and Specific Biogas Production

Total and Specific Methane Yields
Methane Concentration
Gas Production rate over the incubation period
Methane Production rate over time

Biogas: Biogas is produced by bacteria through the bio-degradation process of organic material under
anaerobic conditions. The composition of the resulting biogas depends upon feed material. Its caloric
value is 20 Mega Joules (MJ) /m3 and it usually burns with 60 % efficiency in a conventional biogas
stove. This gas is useful as fuel to substitute firewood, cow-dung, petrol, LPG, diesel, & electricity,
depending on the nature of the task, and local supply conditions and constraints.

Hydrogen
Methane Carbon Dioxide Water Ammonia Nitrogen Oxygen Hydrogen
Sulphide
(CH4) ( CO2) (H2O) (NH3) (N2) (O2) (H2)
(H2S)
50-75% 25-50% 2-7% < 1% < 1% < 3% < 1% < 1%
Table 1: Biogas composition

Equipment

MilliGascounter: Sartorius MC 210P

Magnetic Stirrer: Heidolph MR3000
pH Meter: Knick (Berlin), ph Meter 761 Calimatic
Oven: WTB Binder ED 115 (115 litres), dry matter content 8h 105C
Incubator: WTB Binder BD 115 with additional PC fan, for the incubation of 250 ml digestion vessels
Digestion vessels: 250 ml glass bottles with bypass
Metal Tube: (OD 7m, length 15cm), with glass beads for mixing of the gas in the bag
Online Methane Sensor: BACVis, Datablogger CounterVis BluSens, Bottrop (www.blusens.de)

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 1

Experimental Methodology:

Generally the entire experiment is consist of two main steps, first is the preparation of the inoculum and
then preparing the vessels (mini-fermenters) and conducting the fermentation process. The seed sludge
was provided from the Geesthacht sewage treatment plant. Total Solid (TS) content in the primary sludge
(raw sludge) is approximately in the range 2-7% (Turovskiy and Mathai, 2006). The minimum TS
content of the inoculum is approximately around 4-4.5% for biogas production. Therefore the raw sludge
must be refined by adding a flocculating agent.

In total 18 fermenters with 250ml total volume are prepared for the experiment. Two different types of
fermenters are used in the experiment; first type has only one opening which is used for analysing the
biogas yield. The other type has two openings, one for measuring the biogas yield and the other for
measuring methane production through methane sensors. Six of the latter fermenters are used to measure
the rate of methane production.

Each of the fermenters is filled with the respective amounts of sludge and substrates as mentioned in the
table (2), and the pH values are taken. In summary fermenters 1-5 contain 100g sewage sludge plus 4g
wheat straw powder, fermenters 6 -10 contain 100g sewage sludge and 60g of Grass Juice Silage,
fermenters 11-15 contain 100g sewage sludge and 2g cellulose, and finally fermenters 16 -18 has only
inoculum which are blank controls. After all the fermenters are prepared, they will be placed inside the
incubator as required and then connected to the measuring devices through the piping system provided.
The temperature of the incubator is maintained at 41C for creating mesophilic conditions. The gas yields
of the fermenters are detected from the MillliGascounter and methane sensors, which are automatically
transmitted into a computer. Over the entire incubation period, manual readings of the instruments are
taken daily for ensuring consistency. During the experiment period an unfortunate event happened that is
the computer located in the laboratory defected and shut down and so computer-recorded data were lost
for the last 19 days of this experiment. Therefore, manual readings are used along with the computer data
and due to this some inconsistencies might appear into the graphs of parameters.

Simultaneously, analysis of the total solid (TS) content and volatile solids (VS) is done. In a previously
weighed aluminium crucible, the sludge sample is weighed and dried at 105C for about 8 hours. Finally
the average of 6 specimens is considered as the dry weight. The VS is the amount of the organic dry
matter in the substrate. For its determination, the dried samples are ignited to constant weight in a muffle
furnace for 5 hours at a temperature of 540C. The VS content is calculated by subtracting the ash from
the TS.

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 2

 Figure 1: Adding flocculating agent Figure 2: Filling the inoculum in the fermenters

Figure 3: Measuring pH value Figure 4: placing the fermenters in the incubators

Source: Hamburg University of Applied Sciences (HAW)

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 3

Composition of Batches:

Table 2 shows the composition of the batches. Five vessels were prepared for each of the three selected
substrates, namely wheat straw powder, Grass Juice Silage and cellulose. This was done to obtain more
samples that are able to closely represent the behaviour of the substrates. Three vessels were used for the
blank input (where no substrate was added) to serve the purpose of measuring the rate of biogas
production of the inoculum.

Composition

No. Trace Elements

No Methane Seed Sludge see Table Trace Substrate
Sensor Elements

IV/1 12448
IV/2
250 ml Vessel 4g
IV/3 100 g Geesthacht Sewage Sludge 5,17 % 1,7 ml Wheat Straw Powder
TS + 20 ml Water
IV/4 12450
IV/5
IV/6
IV/7 12452
250 ml Vessel
60 g
IV/8 100 g Geesthacht Sewage Sludge 5,17 % 1,7 ml
Grass Juice Silage
TS
IV/9
IV/10 12454
IV/11
IV/12
250 ml Vessel
2g
IV/13 12456 100 g Geesthacht Sewage Sludge 5,17 % 1,7 ml
Cellulose
TS
IV/14
IV/15
IV/16 12458
250 ml Vessel
IV/17 100 g Geesthacht Sewage Sludge 5,17 % 1,7 ml Blank
TS
IV/18

Table 2: Overview of the composition of batches for fermentation in Incubator IV

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 4

pH changes for the Sewage Sludge and Substrates:

The pH values were measured before and after fermentation for all the batches.
Composition pH

No.
pH Start pH End
No Methane Seed Sludge Substrate
13.11.15 11.12.15
Sensor

IV/1 12448 7.91 7.27

IV/2 4g 7.82 7.22

250 ml Vessel
Wheat Straw
IV/3 100 g Geesthacht Sewage 7.87 7.22
Powder
Sludge 5,17 % TS
IV/4 12450 + 20 ml Water 7.94 7.24

IV/5 7.90 7.27

IV/6 6.16 7.60

IV/7 12452 6.13 7.65

250 ml Vessel
60 g
IV/8 100 g Geesthacht Sewage 6.04 7.63
Grass Juice Silage
Sludge 5,17 % TS
IV/9 6.08 7.63

IV/10 12454 6.02 7.65

IV/11 7.92 7.35

IV/12 8.01 7.32

250 ml Vessel
2g
IV/13 12456 100 g Geesthacht Sewage 8.10 7.32
Pure Test Cellulose
Sludge 5,17 % TS
IV/14 8.00 7.34

IV/15 7.95 7.31

IV/16 12458 8.03 7.53

250 ml Vessel
IV/17 100 g Geesthacht Sewage Blank 7.97 7.46
Sludge 5,17 % TS
IV/18 7.91 7.46
Table 3: pH values of all batches of Incubator IV before and after fermentation

Comments: As seen in the table, the pH values decrease after fermentation for all substrates, except
Grass Juice Silage. The pH values for the other substrates, except Grass Juice Silage, are slightly higher
than 7 before fermentation which indicates alkaline nature. This is because Grass Juice Silage undergoes
a pre-fermentation stage where a lowering of pH is observed.

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 5

Determination of Dry Matter Content Organics (VS) and Degradable Organics (VSf) of the
Substrates:

In the laboratory, the total solids (TS) and volatile solids (VS) were determined by first placing the
samples in aluminium crucibles for 8 hours at 105oC to get the TS content, followed by igniting the dry
matter in a muffle furnace for 5 hours at 540oC to get the VS content. The changes in weights from wet
weight (ww) to dry weight and loss of ignition are used to calculate TS and VS as follows:

Formula to calculate TS (Results are displayed in Table 4):

TS [%] = * 100

Formula to calculate VS (Results are displayed in Table 4):

VS [%] = * 100

To calculate the biogas production per gram of degradable organics (VSf), the following formula is used:

( )

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 6

Results:

The biogas measurements were carried out by an online system named BACCom Data every two hours
per day. Manual measurements were also taken twice a day (roughly at 10:00 and 16:00) as a safeguard
to prevent data loss in case of a failure of the online system. The biogas measurements are shown in
Table 4.

IV. Biogas Station, with MGCs and Methane Sensors, 41 C

Composition pH BACCom Data + MGC-Manual List
Trace
No.
Elements pH pH
Methan gros ml MGC grosNml average netNml
No Seed Sludge see Table Substrate Start13.11.1 End11.12.1
e (34 days) (STP) value (STP)
Trace 5 5
Sensor Elements
IV/1 12448 7.91 7.27 2455.65 2234.64
250 ml Vessel 4g
IV/2 100 g Wheat Straw 7.82 7.22 2477.64 2254.66
IV/3 Geesthacht 1,7 ml Powder 7.87 7.22 2498.04 2273.22 2188.83 1825.39
IV/4 12450 Sewage Sludge + 20 ml 7.94 7.24 2507.09 2282.15
5,17 % TS Water
IV/5 7.90 7.27 2087.34 1899.48
IV/6 6.16 7.60 2001.33 1821.21
250 ml Vessel
IV/7 12452 100 g 60 g 6.13 7.65 2168.64 1973.46
IV/8 Geesthacht 1,7 ml Grass Juice 6.04 7.63 2286.94 2081.12 1924.18 1560.74
IV/9 Sewage Sludge Silage 6.08 7.63 2021.39 1839.46
5,17 % TS
IV/10 12454 6.02 7.65 2094.13 1905.66
IV/11 7.92 7.35 2174.40 1978.71
250 ml Vessel
IV/12 100 g 8.01 7.32 2045.48 1861.57
2g
IV/13 12456 Geesthacht 1,7 ml 8.10 7.32 17,63,484 1604.77 1776.61 1413.17
Cellulose
IV/14 Sewage Sludge 8.00 7.34 2015.52 1834.12
5,17 % TS
IV/15 7.95 7.31 1762.50 1603.88
IV/16 12458 250 ml Vessel 8.03 7.53 414.69 377.37
100 g
IV/17 7.97 7.46 388.48 353.51
Geesthacht 1,7 ml Blanc 363.44 363.44
IV/18 Sewage Sludge 7.91 7.46 395.00 359.45
5,17 % TS
Table 4: Manual and digital measurement values of the Test for Incubator IV

The digital values are already given in Norm-mL [mLN], which is the volume at standard temperature and
pressure (STP) condition. However, the manual readings have to be converted to Norm-mL [mLN] by
using the following formula:

( )
[ ]
( )

with:

= 1013 hPa
= 25.29 hPa
= 6 hPa
= 273.15 K
( ) = 294.15 K.

After putting in all the constant parameters in the formula, we reduce the equation and get a conversion
factor of:

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 7

 1. Gross Biogas Production

Gross Biogas Production for Wheat Straw Powder

2500
IV/1-4
Gross Biogas Yield [mLSTP ]
2000

1500

1000

500
IV/16-18

0
0 5 10 15 20 25 30 35 40
Time [d]

Figure (5): Gross biogas production for wheat straw powder (4g) samples (---- IV/1, ----IV/2, ----IV/3, ----IV/4) and sewage
sludge Geesthacht Blank (100g) (-- --IV/16, -- --IV/17, -- --IV/18) from MGC station IV. Anaerobic fermentation test for 34
C with 100g sewage sludge from Geesthacht conditioned at 5.17% TS. Outlier (IV/5) is excluded.

Comments: Biogas production slows down a little in the 9th day. All of the samples follow the same
nature except (IV/5) which is the outlier since it deviated largely from other curves and thus has been
cancelled. It produces least amount of biogas (1821.21mLSTP), compared to other samples a minimum
difference of (78.3mLSTP) can be found.

Gross Biogas Production for Grass Juice Silage

2500
Gross Biogas Yield [mLSTP ]

2000 IV/6-10

1500

1000

500
IV/16-18

0
0 5 10 15 20 25 30 35 40
Time [d]

Figure (6): Gross biogas production for Grass Juice Silage (60g) samples (---- IV/6, ----IV/7, ----IV/8, ----IV/9, and ----IV/10)
and sewage sludge Geesthacht Blank (100g) (-- --IV/16, -- --IV/17, -- --IV/18) from MGC station IV. Anaerobic fermentation
test for 34 days at 41 C with 100g sewage sludge from Geesthacht conditioned at 5.17% TS.

Comments: Biogas production is normal in the 1st day for all the samples except (IV/6) which is slightly
slower in the beginning. In the 2nd day all samples see a small decrease but strive to increase back quickly
until 10th day. However, after that the production rate starts to decrease and approaches zero in the last
day.

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 8

 Gross Biogas Production for Pure Test Cellulose
2500

2000
IV/11-15
Gross Biogas Yield [ mLSTP ]

1500

1000

500
IV/16-18

0
0 5 10 15 20 25 30 35 40
Time [d]

Figure (7): Gross biogas production for pure test cellulose (2g) samples (---- IV/11, ----IV/12, ----IV/13, ----IV/14, and ----
IV/15) and sewage sludge Geesthacht Blank (100g) (-- --IV/16, -- --IV/17, -- --IV/18) from MGC station IV. Anaerobic
fermentation test for 34 days at 41 C with 100g sewage sludge from Geesthacht conditioned at 5.17% TS.

Comments: All samples follow the same nature but some are more spread from others. Biogas
production rate is very high during in the first two days but then it slows quite fast. Sample IV/11 and
IV/15 produce the maximum (1941.1mLSTP) and minimum (1560.74mLSTP) amount of gas respectively,
but IV/12, IV/13 and IV/14 are in the average zone.

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 9

 2. Net Biogas Production

Biogas Production Curves (Mean net values) from Wheat Straw Powder,
Grass Juice Silage and Pure Test Cellulose
2000
1800 A
1600
Net Biogas Yield [mLSTP ]

B
1400 C
1200
1000
800
600
400
200
0
0 5 10 15 20 25 30 35 40
Time [d]

Figure (8): Net biogas production for wheat straw powder A (----), grass juice silage B (----), and cellulose C (----), from
MGC station IV. Anaerobic fermentation test for 34 days at 41 C with 100g sewage sludge from Geesthacht conditioned at
5.17% TS.

Comments: As indicated in the graph, wheat straw powder produced the highest amount of biogas
(2188.82 mLSTP) although it started slowly compared to Grass Juice Silage and cellulose. Grass Juice
Silage has the second largest production (1924.2mLSTP), the production rate is very high in the 1st day
until it gradually reaches to zero in the 13th day. Cellulose produces the minimum amount of biogas
(1776.61mLSTP).
Biogas yield [NmL] related to 1g of degradable organic matter in substrates

Input TS VS VS Lignin Degradable Substrate

Content Content (Not Corrected) /TS VS /Assay
Geesthacht Sewage Sludge
8.79 % 5.68 % 0.0568 - - -
10.11.15
BiEn Grass Juice Silage
5.15 % 3.84 % 0.0384 0.36 0.0329 1.974
10.11.15
Wheat Straw Powder 98 % 90 % 0.882 0.25 0.637 2.548
Pure Test Cellulose 98 % 100 % 0.98 0 0 0
Table 5: Properties of Sewage Sludge and Substrates

The total solids (TS) and volatile solids (VS) were computed after drying the wet weight (ww) and then
igniting it respectively.

The following formula was used to calculate the degradable content:

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 10

 Gross Biogas yield per 1g of
Net Nml Mass of Net Nml (STP) / g
No. Substrate Nml substrate, Nml/g TS of
average substrate degradable VS
average substrate
IV/1
IV/2 Wheat
IV/3 Straw 2188.83 1825.39 4g TS 456.347 716.248
IV/4 Powder
IV/5
IV/6
BiEn
IV/7
Grass
IV/8 1924.18 1560.74 60g ww 26.012 790.273
Juice
IV/9
Silage
IV/10
IV/11
IV/12
Pure Test
IV/13 1776.61 1413.17 2g TS 706.585 720.918
Cellulose
IV/14
IV/15
Table 6: Specific biogas yield values

Net Biogas Production per Gram on Average

2000 1825
1560
1413
Biogas Yield [mLSTP]

1500

1000 790.273
716.248 677.083 706.5 720.918 720.918
456.25 517.29
500

26
0
Net BGP/ BGP/ 1 g BGP/ 1 g BGP/1 g Net BGP/ BGP/ 1 g BGP/ 1 g BGP/1 g Net BGP/ BGP/ 1 g BGP/ 1 g BGP/1 g
Assay dw VSf VS Assay ww VSf VS Assay ww VSf VS
Wheat Straw Powder Grass Juice Silage Pure Test Cellulose
Wheat Straw Powder Net BGP/ Assay Wheat Straw Powder BGP/ 1 g dw
Wheat Straw Powder BGP/ 1 g VSf Wheat Straw Powder BGP/1 g VS
Grass Juice Silage Net BGP/ Assay Grass Juice Silage BGP/ 1 g ww
Grass Juice Silage BGP/ 1 g VSf Grass Juice Silage BGP/1 g VS
Pure Test Cellulose Net BGP/ Assay Pure Test Cellulose BGP/ 1 g ww
Pure Test Cellulose BGP/ 1 g VSf Pure Test Cellulose BGP/1 g VS

Figure (9): Net specific biogas production on average. VSf = Degradable Organics, ww = Wet Weight

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 11

Description: The first column was computed with the total average net biogas yield. The second column
for all three substrates is the biogas yield per gram wet weight. This is nothing but the total average net
biogas yield divided by 4 for wheat straw powder, by 60 for the Grass Juice Silage and by 2 for cellulose.
The third column depicts the biogas yield per gram of fermentable organics (FO). While calculating
fermentable organics, it must be noted that wheat straw powder and Grass Juice Silage have a lignin
content that cannot be degraded. Therefore this content should be subtracted from the VS. In case of the
Grass Juice Silage, there are volatile fatty acids (VFA) that contribute to the biogas and thus need to be
added.

( )

The biogas production of wheat straw powder per gram of degradable organics is calculated as follows:

716.248 mL
( )

The biogas production of Grass Juice Silage per gram of degradable organics is calculated as follows
(VFA content in WW is 1.32 %):

790.273 mL
( )

The biogas production of cellulose per gram of degradable organics is calculated as follows:

720.918 mL
( )

Degradation Rate of Cellulose

As the molecular formula, molecular weight and the carbon content of cellulose (C6H10O5) are known, a
theoretical calculation of the total gas volume that can be obtained using cellulose can be made.
According to the gas law, 1 mole of carbon has a potential norm gas volume of 22.413968 L (Scherer,
2013). The amount of carbon molecules available for the generation of gas are 35.12625 mmol per g of
cellulose. The theoretical total gas volume using 2 g of cellulose can be calculated as:

The difference between the experimental and theoretical gas yields are shown in the table below:

VFinal Gas Yield [mL] Vtheoretical [mL] Difference [mL] Degradation

1413 1574.637 161.637 89.735 %
Table 7: Degradation rate of cellulose

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 12

 Standard Deviations of the Specific Biogas Production per Assay
3000
Blank Biogas Production
Net Biogas Production
2500
145NmL, 8%

95NmL, 6%
2000 148NmL, 11.5%
Biogas Yield [mLSTP]

1500
1825
1560 1413
1000

500

363 363 363

0
Wheat Straw Powder/ Assay Grass Juice Silage/ Assay Pure Test Cellulose / Assay

Figure (10): Net biogas production for wheat straw powder, grass juice silage and pure test cellulose from MGC station IV.
(Average values with deviations considered). Anaerobic fermentation test for 34 days at 41 C with 100g sewage sludge from
Geesthacht conditioned at 5.17% TS. Outlier accounts for 0.3% deviation.

Comments: The net biogas production for each of the substrates is shown in their respective columns.
Wheat straw powder has produced the maximum amount of biogas (1825mLSTP 145), and the least
amount comes from cellulose (1413mLSTP 148) with biggest deviation. In summary, wheat straw
powder has the highest potential for biogas production.

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 13

 Standard Deviation (Wheat Straw Powder)
2500

2000
Biogas Yield [mLSTP]

1500

1000

500

0
0 5 10 15 20 25 30 35
Time [d]

Figure (11): Net biogas production for wheat straw powder from MGC station IV. (Average values with deviations: 146).
Anaerobic fermentation test for 34 days at 41 C with 100g sewage sludge from Geesthacht conditioned at 5.17% TS.

Wheat straw powder yielded a total amount of (2188.82mLSTP) biogas during the entire 35-days and it
has the second largest deviation ( 146mLSTP) among all the substrates, slightly lower than cellulose, as
indicated by the air-band shadings. For a direct comparison of all the substrates, refer to figure (10).

2500 Standard Deviation (Grass Juice Silage)

2000
Biogas Yield [mLSTP]

1500

1000

500

0
0 5 10 15 20 25 30 35
Time [d]

Figure (12): Net biogas production for Grass Juice Silage from MGC station IV. (Average values with deviations: 95).
Anaerobic fermentation test for 34 days at 41 C with 100g sewage sludge from Geesthacht conditioned at 5.17% TS.

Comments: Grass Juice Silage yielded a total amount of (1924.2mLSTP) biogas during the entire process
and it has the least deviation ( 95mLSTP) among the substrates, very different from the straw powder (
146mLSTP) and cellulose ( 148mLSTP). For a direct comparison of all the substrates, refer to figure (10).

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 14

 2500 Standard Deviation (Cellulose)

2000

1500
Biogas Yield [mLSTP]

1000

500

0
0 5 10 15 20 25 30 35
Time [d]

Figure (13): Net biogas production for pure test cellulose from MGC station IV. (Average values with deviations: 148).
Anaerobic fermentation test for 34 days at 41 C with 100g sewage sludge from Geesthacht conditioned at 5.17% TS.

Comments: Cellulose yielded the lowest amount of biogas (1776.61mLSTP) compared to wheat straw
powder (2188.8mLSTP) and Grass Juice Silage (1924.2mLSTP). Furthermore, cellulose has the highest
deviation (( 148mLSTP) as well. For a direct comparison of all the substrates, refer to figure (10).

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 15

Exemplary Calculations with real values of gas production

I/S Ratio

The I/S Ratio is the ratio of the organic dry weight of the inoculum over that of the substrate. The
percentage of the organic dry weights (VS) of the inoculums and the substrates are taken from Table 4 for
the following calculations.

I/S Ratio Values

Wheat Straw Powder 2.229
Grass Juice Silage 2.877
Pure Test Cellulose 2.898
Table 8: I/S Ratio

For wheat straw powder, the I/S Ratio is calculated as:

( )

For Grass Juice Silage, the I/S Ratio is calculated as:

( )

For cellulose, the I/S Ratio is calculated as:

( )

Previously conducted experiments have proven that the highest gas yields can be achieved when the I/S
Ratio is between 2 to 2.5 (Scherer 2013). The I/S Ratio for Grass Juice Silage and cellulose is much
higher than the aforementioned range. Thus, less substrate should be used to obtain the ideal I/S Ratio.

Gas Quality (Methane Concentration)

The gas quality is calculated as:

( ( ))

The methane concentration of the produced gas changes over time and thus needs to be calculated as
follows:

( )

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 16

 ( ( ))

( ( ))

( )

When comparing the Gas Quality of the three substrates, it can be seen that cellulose has a lower Gas
Quality as compared to wheat straw powder and Grass Juice Silage.

Parameter Wheat Straw Grass Juice Pure Test Sewage Sludge

Powder (4g) Silage (60g) Cellulose (2g) Geesthacht
Gross Methane Yield (Nml) 1138.774 1163.204 742.1046 127.5595
Gross Biogas Yield (Nml) 2258.397 1939.561 1604.77 377.3652
Average Methane 50.424 59.972 46.244 33.803
Concentration (%) (52) (50)
Table 9: Methane concentrations per batch assays

Net Methane Yield per Gram on Average

2000
1800
Methane Yield [mLSTP]

1600
1400 1163.204
1138.774
1200
1000
742.105
800
600 473.942 406.06
372.449 353.25 360.459 360.459
400 237.25 268.991
200 15.593
0
NmL / NmL/ 1 g NmL/ 1 g NmL/1 g NmL/ NmL/ 1 g NmL/ 1 g NmL/1 g NmL/ NmL/ 1 g NmL/ 1 g NmL/1 g
Assay dw VSf VS Assay ww VSf VS Assay ww VSf VS
Wheat Straw Powder Grass Juice Silage Pure Test Cellulose
Wheat Straw Powder NmL / Assay Wheat Straw Powder NmL/ 1 g dw
Wheat Straw Powder NmL/ 1 g VSf Wheat Straw Powder NmL/1 g VS
Grass Juice Silage NmL/ Assay Grass Juice Silage NmL/ 1 g ww
Grass Juice Silage NmL/ 1 g VSf Grass Juice Silage NmL/1 g VS
Pure Test Cellulose NmL/ Assay Pure Test Cellulose NmL/ 1 g ww
Pure Test Cellulose NmL/ 1 g VSf Pure Test Cellulose NmL/1 g VS

Figure (14): Net specific methane yield per gram on average. VSf = Degradable Organics, ww = Wet Weight, dw = Dry
Weight

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 17

Methane Production

CH4 Concentration [ % ]: Wheat Straw Powder, Grass Juice Silage, Pure Test
Cellulose and Sewage Sludge Geesthacht
90
80
Methane Percentage (%)

70
A, B
60
50 C
40
30
D
20
10
0
-10 0 5 10 15 20 25 30 35 40
Time [d]

Figure (15): Methane concentration for Wheat Straw Powder A (----), Grass Juice Silage B (----), Pure Test Cellulose C (---
-) and Sewage Sludge D (----)) from MGC station IV. Anaerobic fermentation test for 34 days at 41 C with 100g sewage
sludge from Geesthacht conditioned at 5.17% TS.

Comments: Methane content of the Grass Juice Silage is the highest nearly the entire time with a peak
value of 80%. Both wheat straw powder and cellulose see a drop in methane content in the beginning but
they continue to increase back.

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 18

Net Methane Production Rate

Wheat Straw Powder

30

25

20
Methane Yield [ mLSTP]

15

10 *

0
0 5 10 15 20 25 30 35 40
-5
Time [d]

Figure (16): Methane production rate for wheat straw powder from MGC station IV. Anaerobic fermentation test for 34 days
at 41 C with 100g sewage sludge from Geesthacht and 4g Grass Juice Silage, conditioned at 5.17% TS.
* After 16th day measurement values were coupled with manual readings (taken every day) instead of 12 measurements per
day

Comments: In the 2nd day, there is the maximum methane production rate nearly (25mLSTP). From 5th
day to 15, at first the production rate slows down and revolves around an average amount of (4mL STP)
and then gradually decreases to roughly (1mLSTP). In comparison with other substrates, wheat straw
powder has the second-largest methane production rate in this experiment.

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 19

 Grass Juice Silage
18
16
Methane Yield [ mLSTP ]

14
12
10
8 *
6
4
2
0
-2 0 5 10 15 20 25 30 35 40
Time [d]

Figure (17): Methane production rate for Grass Juice Silage from MGC station IV. Anaerobic fermentation test for 34 days
at 41 C with 100g sewage sludge from Geesthacht and 60g Grass Juice Silage, conditioned at 5.17% TS.
* After 16th day measurement values were coupled with manual readings (taken every day) instead of 12 measurements per
day
Comments: During the first two days, the methane production rate increases to approximately (11mLSTP)
first and then decreases back to (5mLSTP). From 4th day to 8th the production rate reaches it is peak value
around (16mLSTP). Again after day 16 the curve follows a different pattern. Based on graphical data,
Grass Juice Silage has the minimum methane production rate among the substrates and thus least
suitable.

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 20

 Pure Test Cellulose
60

50
Methane Yield [ mLSTP ]
40

30
*
20

10

0
0 5 10 15 20 25 30 35 40
-10
TIME [d]

Figure (18): Methane production rate of pure test cellulose from MGC station IV. Anaerobic fermentation test for 34 days at
41 C with 100g sewage sludge from Geesthacht and 2g cellulose, conditioned at 5.17% TS. * After 16th day measurement
values were coupled with manual readings (taken every day) instead of 12 measurements per day
Comments: Starting from the 1st day, the methane production rate has a sharp increase reaching a
maximum amount of (55mLSTP) and in the 3rd, the production rate decreases dramatically to roughly
(0.4mLSTP). From 3rd day to 14th, it is very slow and in the 16th day there is a drop which marks manual
data addition. As can be seen from the graphs, cellulose has the maximum potential for producing
methane. Therefore, among all the substrates, it is the best and most suitable for producing methane.

Sewage Sludge Geesthacht

7

6
Methane Yield [ mL STP ]

3 *
2

0
0 5 10 15 20 25 30 35 40
-1
TIME [d]

Figure (14): Methane production rate for the sewage sludge Geesthacht (Blank) from MGC station IV. Anaerobic
fermentation test for 34 days at 41 C with 100g sewage sludge from Geesthacht conditioned at 5.17% TS. * After 16th day
measurement values were coupled with manual readings (taken every day) instead of 12 measurements per day
Comments: The purpose of conducting the fermentation test only from sludge is just to examine how
methane production will be and what nature it follows. The sludge curve graphed here could be a
reference for comparing the methane production rate with other substrates. In the first 5 day, the methane
production rate is very low around (2.3mLSTP). The 16th day sees a sharp increase approximately
(6.2mLSTP).

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 21

Conclusion
We can conclude from the degradation (89.735 %) between the theoretical and practical gas yield of
cellulose that the setup of the mini fermentation test is very good. A result from the practical course is
that the cellulose is digested the quickest, for the most part after five days. Moreover, the cellulose has
the highest specific gas yield of digestible matter of 706.5 Nml. Another important factor to highlight is
that the Grass Juice Silage has an extremely low specific gas yield of total matter of 26 Nml [Figure 9].
This is due to a very content of non-digestible organic matter (Lignin).

Biogas Engineering Practical Course | Hamburg University of Applied Sciences (HAW) 22