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Energy Procedia 50 (2014) 702 710

The International Conference on Technologies and Materials for Renewable Energy, Environment
and Sustainability, TMREES14

Production of Hydrogen and Methane from Banana Peel by

Two Phase Anaerobic Fermentation
Chananchida Nathoaa, Ubonrat Sirisukpocab, Nipon Pisutpaisala,c, *
a
Department of Agro-Industrial, Food and Environmental Technology, Faculty of Applied Science,
King Mongkuts University of Technology North Bangkok, Bangkok, 10800, Thailand
b
Program of Computer, Faculty of Science and Technology, Nakhon Pathom Rajabhat University,
Nakhon Pathom, 73000, Thailand
c
The Biosensor and Bioelectronics Technology Centre, King Mongkuts University of Technology
Northn Bangkok 10800, Thailand

Abstract

A two phase anaerobic digestion of banana peel is an attractive process for producing hydrogen and methane. Comparative
performance and total energy recovery between two phase process (sequential hydrogen and methane fermentation) and one
phase process (methane fermentation) in were evaluated in batch reactor under mesophilic incubation at varying ratios of
feedstock to microbial inoculum (F/M) ranging from 2.5 10. F/M ratios influence biogas yield, production rate, and potential.
Best performance of one phase methane fermentation was observed at F/M of 5.0. At this condition, methane yield, production
rate, and potential were 251.3 mL g-1 VS, 2.05 mL h-1, and 352.9 mL, respectively. Hydrogen and methane yields of 209.9 and
284.1 mL g-1 VS mL g-1 VS were achieved at F/M of 5.0 in two stage process. Acetic acid is the main volatile fatty acids (VFAs)
produced in the hydrogen fermentation stage. Little amount of VFAs were accumulated in methane fermentation in both stage
processes. Total energy recovery in two stage process is higher than that in one stage by 81%. This work demonstrated two
phase achieved a better performance than one phase fermentation of banana peel.
2014 Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
2014 The Authors. Published by Elsevier Ltd.
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Selection and peer-review under responsibility of the Euro-Mediterranean Institute for Sustainable Development (EUMISD).
Selection and peer-review under responsibility of the Euro-Mediterranean Institute for Sustainable Development (EUMISD)
Keywords: Hydorogen; methane; banana peel; fermentation; two phase process

* Corresponding author. Nipon Pisutpaisal Tel.: +662-587-8257; fax: +662-587-8257.

E-mail address: ubonrat76@hotmail.com, npp@kmutnb.ac.th

1876-6102 2014 Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Selection and peer-review under responsibility of the Euro-Mediterranean Institute for Sustainable Development (EUMISD)
doi:10.1016/j.egypro.2014.06.086
 Chananchida Nathoa et al. / Energy Procedia 50 (2014) 702 710 703

1. Introduction

Hydrogen is one of the most promising alterative to fossil fuels due to its clean, renewable, and high energy yield
(122 kJ g-1). It could be either directly combusted or used to produce electricity through fuel cells. Many processes,
thermochemical, electrochemical, and biological can be used to generate hydrogen [1]. Thermochemical and
electrochemical processes are not renewable due to their dependence on fossil fuels. Biological hydrogen
production more environmentally friendly since it is less energy intensive than non-biological processes.
Biohydrogen can be generated from different metabolic pathways including direct water biophotolysis by green
algae, indirect water bio-photolysis by cyanobacteria, photofermentation by photosynthetic purple non-sulfur
bacteria, hydrogen synthesis via water-gas shift reaction by microalgae in Rhodospirillaceae family, and dark
fermentation by heterotrophic anaerobic bacteria. Higher production rate, wider spectrum of substrates, and lower
energy requirement make dark fermentation more advantageous than other processes [2-4]. Broad types of organic
feedstocks such as glucose, starch, municipal waste, livestock manure, crop residues, food waste, and wastewater
have been utilized as substrates in the dark fermentation [5-8].
Banana is one of the most popular tropical fruits consumed worldwide. In Thailand, banana peel of 200 tons is
daily generated, and its generating trend becomes continuously increased [9]. A small fraction of the peel is used
for animal feed and the remaining becomes rotted garbage. The peel is carbohydrate rich (Table 1) and is potentially
converted into hydrogen and methane in the fermentation process.

Table 1. Organic content in ripped banana peel

Parameter % Wet weight

Moisture 83.5
Organic carbon 41.3
Starch 1.2
Protein 1.8
Cellulose 8.4
Himecellulos 5.3
Glucose 2.4
Total sugar 29

The present study compared the performance of one and two stage mesophilic fermentation of the banana peel based
on biogas yield and production rate, and overall energy recovery.

2. Materials and methods

2.1 Inoculum
Microbial seed used throughout the study was obtained from a full-scale up ow anaerobic sludge blanket
(UASB) reactor treating cassava wastewater (Eiamburapa Co., Ltd., Thailand). Coarse matter >0.5 mm diameter
was removed by sieving and the granules were washed twice with tap water. The fine granules, used in the hydrogen
fermentation were boiled at 100oC for 30 min to deactivate methanogens [10], while those without heat treatment
were used in methane production stage.

2.2 Banana peel

Ripped banana peel was obtained from local banana processing store. It was rinsed with tap water and ground in a
blender to particles of diameter approx. 0.5 mm. The food waste was adjusted to the desire concentration (g VS L-1)
with distilled water.
704 Chananchida Nathoa et al. / Energy Procedia 50 (2014) 702 710

2.3 Batch fermentation

The two stage batch fermentation system including H2 fermentation in the first stage and CH4 fermentation in the
second stage was set up in 500 mL screw-cap bottles with a working volume of 500 mL (375 mL of feed stock and
125 mL of inoculum). The inoculum in all conditions was fixed at 4.75 g total volatile solids (VS) per batch. The
banana peel was added at varying concentrations between 23.75-95.0 g VS L-1, corresponding to F/M ratios of 2.5-
10. The initial pH in the H2 fermentation was adjusted to 6.0 with 6 N NaOH or 6 N HCl. The system was flushed
with nitrogen gas to generate anaerobic conditions. H2 fermentation was conducted at 37C with rotary shaking at
150 rpm. After H2 fermentation was exhausted, the non-heat pretreated seed of 4.75 g VS per batch was added, the
initial pH was adjusted to 7.0, and the system was reflused with nitrogen gas, and the CH4 fermentation was
conducted at 37C. All experiments were setup in duplicate. During the fermentation experiment, total gas volume
and composition were periodically monitored by gas counters and gas chromatography, respectively. The liquid
samples were analyzed for pH and volatile fatty acids (VFA) every 4-6 h.

2.4 Analytical methods

Total solids (TS), total volatile solids (VS), and chemical oxygen demand (COD) were measured according to
Standard Methods 2540 G and 5220 B, respectively [11]. The amount of generated biogas was recorded using
liquid displacement gasometers. Biogas content (H2, CH4, and CO2) was measured periodically every 5 h using a
gas chromatograph (Shimadzu GC-8A, Kyoto, Japan) equipped with a thermal conductivity detector (TCD) with a
Unibeads C 60/80 column (GL Sciences, Inc., Tokyo, Japan). Helium was used as a carrier gas. The temperatures
of the injection port and the detector were 150 and 80C, respectively. VFAs were analyzed by gas chromatography
(Shimadzu GC-7A system equipped with a flame ionization detector and a Stabilwax DA capillary column (Restek
Corporation, PA, USA)). The temperatures of the injection port and detector were maintained at 240C [12].

2.5 Kinetics of gas production

The modified Gompertz equation (eqn. 1) was used to fit cumulative hydrogen/methane production data obtained
from each batch experiment. This model has long been used for describing hydrogen, methane, or biogas
production in batch fermentation experiments.

R e
H (t ) P exp  exp m (O  t )  1 (1)
P
where H(t) is cumulative hydrogen production (mL) during the incubation time, t (h), P is the biogas production
potential (mL), Rm is the maximum production rate (mL h-1), is the lag phase duration (h), and e is the exp(1) =
2.718. Biogas yield (Y) is calculated by dividing the biogas production potential by the amount of VS removed.

3. Results and Discussions

3.1 One stage fermentation

pH in the reactor during one stage banana peel fermentation at varying F/M ratio in the range of 2.5 10 was
stable in the range of 6.72 7.30 (Fig. 1). The results implied that the banana peel might be composed of buffer
compounds that can maintain the fermentation pH to suit the methanogenic activity. Methane was produced after 12
h incubation for all F/M ratios with the methane content approximately4854 % and the accumulative methane
reached the plateau region after 240 h fermentation (Fig. 2). The cumulative methane profile data was fitted to the
modified Gompertz equation (R2 > 0.99). The methane evolution had the S-shape trend and the kinetics data from
the equation was statistically significant. Methane production rate and yield were increased as the increased F/M
ratios and maximized at F/M of 5.0 (Fig. 3). The kinetics parameters were decreased when the F/M ratio above 5.0
(Table 2). Methane yield obtained from the present study was comparable to those in the previous studies (Table 3).
Little amount of VFA (< 5 mM) accumulated in the reactor content suggested that no inhibitory effect of VFA on
the microorganisms.
 Chananchida Nathoa et al. / Energy Procedia 50 (2014) 702 710 705

8.0

7.5

7.0

6.5
pH

6.0

5.5

5.0
0 50 100 150 200 250 300

Time (h)

Fig. 1. pH during one stage fermentation under varying F/M ratio of 2.5 (U), 5 (), 7.5 ({), 10 (S). Symbols represent mean values of triplicate
experiments, error bars represent one standard deviation.

400
350

300

250
Cumulative CH4 (mL)

200

150

100

50

0
0 50 100 150 200 250 300
Time (h)

Fig. 2. Cumulative methane production from F/M ratios of 2.5 (U), 5 (), 7.5 ({), 10 (S). Symbols represent mean values of triplicate
experiments, error bars represent one standard deviation.
706 Chananchida Nathoa et al. / Energy Procedia 50 (2014) 702 710

Table 2. Summary of kinetics parameters in one stage methane production.

F/M Hm Rm Y % pH [VFA]
ratio (mL) (mL h-1 ) (mL g-1VS) mmol L-1 (h)
2.5 277.3 29 1.54 0.3 250.129 54 7.20.1 0.270.05 7.6
5.0 352.8 31 2.0 0.7 251.3 10 47 7.10.1 0.240.04 7.6
7.5 223.4 15 1.4 0.1 134.5 3 33 7.30.1 0.380.10 10.9
10.0 85.1 17 1.2 0.1 38.1 8 31 7.30.1 2.880.09 12.1

5300
A B
250
4
m (mL h )
-1

200
3
Y (mL g RTVS)

150
-1

2
100
1
50

0
0
2.5 5.0 7.5 10.0
300 2.5 5.0 F/M Raito 7.5 10.0
F/M ratio
B
250

200
Y (mL g TVS)

150
-1

100

50

0
2.5 5.0 7.5 10.0
F/M ratio

Fig. 3. Methane production rate (A) and yield (B) under varying F/M ratios. Histograms represent mean values of triplicate experiments, error
bars represent one standard deviation.
 Chananchida Nathoa et al. / Energy Procedia 50 (2014) 702 710 707

Table 3. Methane yield reported by previous studies

Waste Reactor Yield Reference

mL CH4 g-1 VS
Banana peel Batch 210 [13]
Banana peel Batch 266 [14]
Banana peel Batch 231 [15]
Banana peel Batch 251 This study

3.2 Two stage fermentation

pH in the hydrogen fermentation (first stage) slightly decreased and stabilized at 5.4 until the end of
fermentation, while that in the methane fermentation (second stage) was stabilized at the narrow range of 7.11
7.22 (Fig. 5). Those stabilized pH ranges are optimum for hydrogen- and methane-producing bacteria. Since no
buffer was added into the reactor, the results suggested that the stabilized pH in both stages was due to the buffer
capacity from the banana peel.

7
pH

5
Hydrogen Methane

4
0 50 100 150 200 250 300

Time (h)

Fig. 5. pH profile during two stage fermentation of F/M 5. Symbols represent mean values of quadruplicate experiments,
error bars represent one standard deviation.

The cumulative H2 and CH4 fermentation profile data were fitted to the modified Gompertz equation (R2 > 0.99).
The hydrogen evolution had the S-shape trend and the kinetics data from the equation was statistically significant.
Lag phase approximately 12 h prior to the evolution of H2 was observed in the first stage fermentation (Fig. 6). No
methane was detected in this stage.
708 Chananchida Nathoa et al. / Energy Procedia 50 (2014) 702 710

100 800
Hydrogen Methane
80
600

Cumulative gas (mL)

Gas content (%)

60
400
40

200
20

0 0
0 50 100 150 200 250 300

Time (h)

Fig. 6. Profile of gas content () and cumulative gas volume (z) during two stage fermentation of F/M 5. Symbols represent mean values of
quadruplicate experiments, error bars represent one standard deviation.

The hydrogen yield and content of 209.9 mL g- 1VS and 35% were achieved after 12 h fermentation (Fig. 6, Fig.
7A). The results showed that the F/M ratios have strong effect on the H2 production. The methane yield and
content in the second stage were 284.12 mL g-1 VS and 61% (Fig. 7B). The analysis of VFA after 50 h fermentation
showed that acetate was the major fermentation by-product with 65 mM. The acetate dominant by-product
suggested the desirable condition for the hydrogen fermentation. Theoretically, maximum hydrogen yield achieved
when acetate-related pathways are the main routes for H2 production that yielded 4 mol H2/ mol glucose. Minimal
ethanol and propionate generated from the hydrogen consuming pathway were detected. Negligible amount of VFA
detected in the methane fermentation stage after 336 h fermentation indicated that VFA produced in the hydrogen
stage was efficiently converted to methane in the second stage. The overall energy recovery from two stages is
equivalent to 6.9 x 10-3 kW-h, which increased by 81% compared to that recovered from one stage fermentation 3.8
x 10-3 kW-h). The finding suggested that the first stage of hydrogen fermentation plays significant role in the
overall improvement of degradation efficiency and biogas recovery. Separation of combined hydrolysis,
acidogenesis, and acetogenesis in the first stage from the second stage fermentation allows more flexibility in
controlling of fermentation conditions and could be more beneficial for simultaneous treating and extracting
complex organic feedstocks or recalcritant organic matters.
 Chananchida Nathoa et al. / Energy Procedia 50 (2014) 702 710 709

250 8
A

200
6

Y (mL g VS)

Rm (mL h-1)
Hm (mL) 150 Hm
-1
4
100
Y

2
50
Rm

0 0
F/M 5
700 8
B
600

6
500
Y (mL g VS)
Mm (mL)

Rm (mL h )
-1
-1

400
4
Mm
300

200 Y
2

100 Rm

0 0
F/M 5

Fig. 7. Hydrogen (A) and methane (B) production potential, rate, and yield during two stage fermentation of F/M 5. Histograms represent mean
values of quadruplicate experiments, error bars represent one standard deviation.

Table 4. Summary of kinetics parameters in two stage fermentation at F/M 5.

Hydrogen stage
Hm Rm Y % pH [VFA]
(mL) (mL h-1 ) (mL g-1VS) mmol L-1 (h)
93.5 19 5.04 1.3 209.9 28 35 5.430.4 65.80.5 1.5
Methane stage
Mmax Rmax Yield % pH VFA
(mL) (mL h-1 ) (mL g-1VS) mmol L-1 (h)
617.6 21 4.4 0.5 284.1 34 67 7.10.1 6.50.1 1.2
710 Chananchida Nathoa et al. / Energy Procedia 50 (2014) 702 710

4. Conclusion

The current study successfully illustrated more efficient performance in two stage than one stage fermentation of
banana peel regarding the energy recovery. The total energy recovery from sequential hydrogen and methane
fermentation was improved by 81% compared to the one stage fermentation. Two stage process is feasible to
simultaneously treat and extract hydrogen and methane from agricultural waste containing high complex organic
molecules.

Acknowledgements
The authors would like to express their gratitude to the Electricity Generation Authority of Thailand (grant no.
52-2115-031-JOB NO. 518-KMUTNB) for the financial support.

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