The organic agricultural waste as a basic

source of biohydrogen production

Cite as: AIP Conference Proceedings 1711, 080002 (2016); https://doi.org/10.1063/1.4941649
Published Online: 12 February 2016

Lies Sriwuryandari, E. Agung Priantoro, Neni Sintawardani, J. Tri Astuti, Dewi Nilawati, A. Mauliva Hada
Putri, Mamat, Suharwadji Sentana, and T. Sembiring

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AIP Conference Proceedings 1711, 080002 (2016); https://doi.org/10.1063/1.4941649 1711, 080002

© 2016 AIP Publishing LLC.

 The Organic Agricultural Waste as a Basic Source of
Biohydrogen Production

Lies Sriwuryandari1,a), E. Agung Priantoro1, Neni Sintawardani2,b), J. Tri Astuti1,

Dewi Nilawati1, A. Mauliva Hada Putri2, Mamat1, Suharwadji Sentana1, T.
Sembiring1

1Research Center for Physics-Indonesian Institute of Sciences

Jalan Sangkuriang, Kompleks LIPI Bandung,40135
2Research Center for Physics-Indonesian Institute of Sciences
PUSPIPTEK, Serpong, Tangerang Selatan,15310
a)
Corresponding author: lies.sriwuryandari56@gmail.com
b)
yisesehat@yahoo.com

Abstract.Biohydrogen production research was carried out using raw materials of agricultural organic waste that was obtained
from markets around the Bandung city. The organic part, which consisted of agricultural waste material, mainly fruit and
vegetable waste, was crushed and milled using blender. The sludge that produced from milling process was then used as a
substrate for mixed culture microorganism as a raw material to produce biohydrogen. As much as 1.2 kg.day-1 of sludge (4% of
total solid) was fed into bioreactor that had a capacity of 30L. Experiment was done under anaerobic fermentation using bacteria
mixture culture that maintained at pH in the range of 5.6-6.5 and temperature of 25-30oC on semi-continuous mode. Parameters
of analysis include pH, temperature, total solid (TS), organic total solid (OTS), total gas production, and hydrogen gas
production. The results showed that from 4% of substrate resulted 897.86 L of total gas, which contained 660.74 L (73.59%) of
hydrogen gas. The rate of hydrogen production in this study was 11,063 mol.L-1.h-1.

INTRODUCTION
The energy crisis due to the growing demand and the depletion of oil reserves may have an impact on various sectors
of industry, agriculture, transportation, and national energy security. Currently the increasing use of petroleum as a fuel
sources are faced with various problems of limited oil reserves and global warming. About 95% of industrial activities
produce gases contributing directly to an increase in ozone depletion, which come from the combustion of carbon-
containing raw materials, especially from fossil [1, 2]. Two problems, i.e. energy from oil and global warming, have
become national issues. Therefore, it is necessary to find alternative energy that comes from domestic sources or from
waste materials that can be sustained. Biomass from organic waste is one example for a domestic source of potential,
which is convertible into energy with consideration of environmental friendly, renewable, and beneficially [3].
Based on the above description, biohydrogen is one alternative that can be developed. We shall implement
biotechnology withmicro-organisms as active ingredient processing [4]. Anaerobic fermentation of biomass waste can
be used to convert raw materials for the production of biohydrogen. This is useful for alternative energy conversion
since it has less energy input.
Hydrogen gas is an energy source that can be converted into electricity and fuel. It is known as a source of clean
energy and environment-friendly source because it leaves no toxic waste. Therefore, it does not cause the greenhouse
effect, ozone depletion, or acid rain, because the combustion process in the air only leaves water vapor and energy [5,6].
Hydrogen has the highest combustion energy ofall types of fuel ever known in the amount of 143 GJ/ton. Combustion
of energy hydrogen can be easily channeled in fuel cell technology into electrical energy, which can be stored [7].
Manufacture of hydrogen through a process of fermentation is inexpensive and easy, as it can be done at a temperature

International Symposium on Frontier of Applied Physics (ISFAP) 2015

AIP Conf. Proc. 1711, 080002-1–080002-6; doi: 10.1063/1.4941649
© 2016 AIP Publishing LLC 978-0-7354-1358-0/$30.00

080002-1
and ambient pressure [8]. It is also very efficient and can be used as fuel for electric generators. Based on the above
description, a research has been carried out to determine the potency of organic agricultural materials in producing
biohydrogen as an alternative energy. This research is also intended to help addressing the issue of agricultural waste
management to be environmentally friendly.

MATERIALS AND METHODS

Biomass Waste
Initially, the garbage that was obtained from the market in Bandung city was divided into two parts, i.e. organic and
non organic material. The organic part, which consisted of agricultural waste material, mainly fruit and vegetable waste
was crushed and milled using blender. The sludge that produced from milling process (Fig.1) was then used as a
substrate for mixed culture microorganism as a raw material to produce biohydrogen. Feeding of organic agricultural
sludge to bioreactor was 1.2 kg.day-1.

Figure 1.Sludge of organic agricultural waste

Bacterial Mixed Culture and Growth Media

Previously, as much as 10 mL of bacterial suspension from stock culture of mixed bacteria was inoculated into
Erlenmeyer containing of sterilized growth media (capacity 2L). Growth media that was used in this study consisted of
K2HPO4 750; KH2PO4 = 850; (NH4)2SO4 = 40; MgSO4.7H2O = 200; CaCl2.2H2O = 0.75; Co (NO3)2.6H2O = 290; Fe
(NH4)2SO4.6H2O = 10; MnCl2.4H2O = 2.1; Na2MoO4.2H2O = 0.75; H3BO3 = 2.8; Cu (NO3)2.3H2O = 0.04;
Na2EDTA.2H 2O = 2.0; Nicotinic acid = 0.025; and yeast extract = 0003 mg.L-1.This culture was used as inoculum in
the bioreactor for biohydrogen process with sludge of organic agricultural waste material as a substrate. Total usage of
inoculum in bioreactor is 10% (v/v). In order to optimize the production of biohydrogen, the temperature was
maintained in the range of 25-30°C and pH at 5.6-6.5. The adjustment of pH was carried out by adding of NaOH 3N to
increase pH or by adding HCL 3N to decrease pH. It is depending on the pH level of substrate.

Bioreactor
The bioreactor in this study has a capacity of 30 L. Basically, it consists of two main parts, i.e. feeder and digester. It
was equipped with electric pump (0.5 HP) with capacity of 40 L.min-1. This pump is required to feed the substrate
properly into bioreactor. Bioreactor was equipped with electric pump to circulate substrate so it could optimize process.
These bioreactors operated semi-continuously. Bioreactor that used in study was presented in Fig.2.

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 Gas Outlet pH Adjustment

Gas Colector
Gas Meter

Circulating
Pump

Bioreactor Feedstock

Feeding Pump

a b
Figure 2. (a) Photograph and (b) schematic diagram of bioreactor used in study

Sample Analysis
As much as 100 mL per day of sample was required for analysis. It was recovered from the outlet pipe of
bioreactor. Parameters of analysis in this study werediscussed below:

pH and Temperature

pH of sample was measured by using pH-meter (LT Lutron pH-207) and temperature was measured by using
thermometer Hg. Total solid (TS) content was determined gravimetrically. About 25 g (A) of sample was placed at
crucible and weighed, then heated at 105oC in Oven (Memmert 400 D-06060) until a constant weight was obtained
(B). TS content was calculated by using Eq. 1.

B
TS u 100% (1)
A

OTS Content

To determined OTS, dry matter in crucible (TS) was burned using Furnace (Thermolyne 62700) at 550 oC. In
combustion, organic material would be degraded to gas and water. Meanwhile, non-organic substance would be
recovered as residue (ash) in crucible. Ash in crucible was weighed (C), then OTS content was calculated by using
Eq. 2.

BC
OTS u 100% (2)
B

Total Gas Production

Total gas produced from bioreactor was measured daily (every 24 hours) by using Gas Flow Meter (Ritter-
Germany). Total gas could be read from the gas flowmeter.

Hydrogen Gas Production

Hydrogen gas was measured by using gas cylinders (500 mL) and determined by infra red syngas analysis
using A Cubic Gasboard-3100P (Wuhan cubic optoelectronic Japan). Sum of Hydrogen gas molecule that produced
by the fermentation process was calculated with using Eq. 3 according to [13] as described below.

P ˜V
N (3)
R ˜T

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where:
N = Sum of hydrogen gas molecule (mol)
P = Pressure of (atm)
R = Gas constant (0.082)
T = Temperature (K)
V = Volume gas (L)

RESULTS AND DISCUSSION

Bacteria require appropriate environmental conditions for optimal growth, such as nutrition, pH, and
temperature. On this study pH level of fermentation was adjusted at around 5.5. According to the previous study,
hydrogen production rate reached its maximum if pH was in the range of 5.0-6.0. It was suggested that in pH level
of 5.5, we could obtain the highest hydrogen production rate [9]. In this study the bioreactor pH was adjusted to the
optimum, namely 5.6.If it was lower than 5.6 or higher than 7.0 the hydrogen production will be low. In this case,
the growth of methane bacteria increased and consumed the hydrogen produced. Production of hydrogen can be seen
in Fig. 3.

Figure 3. Relation between gas production and pH in the bioreactor during fermentation

As shown in the Fig.3 the total biogas obtained fluctuated until 20 days and decreased significantly until the end
of experiment. At the initial stage, the conditions inside the bioreactor were not yet stable because the incoming
biomass settled and a portion formed small dense clumps causingimproper circulation and inhomogeneous broth
media. In this case, bacteria needed to adapt to the atmosphere in the bioreactor. In the next process, we got
acquisition of gas with total amount of 122.71 L.h-1 at pH range of 5.56. This is in accordance with the previous
studies [5,9,7,11,12], which show that anaerobic fermentation produces biogas at pH between 4.8-6.2, while for
methane and carbon dioxide it is at pH of 6.5-7.8. At low pH, bacteria would degrade soluble compounds and
produced several short chains of organic fatty acids such as butyrate, propionate, acetate, and lactate. Afterward,
these organic fatty acids would be degraded by bacteria to produce gas, mainly hydrogen gas and carbon dioxide.
The formation of hydrogen gas would occur optimally at mesophilic temperature of 25 - 30°C and at pH of 5.56.
Controlling pH value is required to maintain the stability of fermentation process. In this study, pH was controlled
around 5.5 (with initial pH 4.10) by adding NaOH 3N. The gas production was decreased to 0.21 L.h-1 at the end of
experiment.
The decreasing of gas production might be caused by the decreasing of organic matter content as nutrient source
for microorganism. Organic matter was used as source of microorganism for metabolism activity to produce biogas.
Therefore, the addition of organic matter is required to prevent growth inhibition and death of microorganism. Fig. 4
shows the effect of organic material to the gas production during fermentation process.

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 Figure 4. Relation between organic total solids and gas production during fermentation

It can be seen in Figure 4 that the content of organic substrates from waste or organic materials tends to decrease,
even takes place slowly. Organic matter is a source of carbon and essential nutrients needed by micro-organisms. It
used as a source of energy for bacteria metabolism during fermentation process. In this study, 1.2 kg substrate was
provided as input into the bioreactor containing 66.85% organic materials. From the total of 897.86 L gas, hydrogen
recovery as much as 660.77 L or equivalent with 73.59% of the total gas was produced (Fig. 5).

Figure 5. Production of total gas and hydrogen during fermentation

Based on the Equation 3 above, it was concluded that the production rate of biohydrogen gas in this study was
11.063 mol.L-1.h-1.

CONCLUSIONS
It can be concluded that agricultural waste (vegetables and fruits) can be used as the basic materials for
producing biogas hydrogen by an anaerobic fermentation using bacterial consortium. They produced 660.77 L of H2
out of 897.86 L total gas obtained (73.59%), with a production rate of 11.063 mol.L-1.h-1. This technique could be
applied to reduce, use, and solve the organic agricultural waste (garbage) problems.

ACKNOWLEDGEMENTS
This research was funded by Research Center for Physics LIPI (DIPA Fiscal Year 2015). We thank to
the Head of Research Center for Physics LIPI, all researchers, technicians, and others who have contributed to
this research.

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