International Journal of Hydrogen Energy 31 (2006) 1284 1291 www.elsevier.

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Biohydrogen generation from palm oil mill efuent using anaerobic contact lter
Krishnan Vijayaraghavan , Desa Ahmad
Department of Biological and Agricultural Engineering, Faculty of Engineering, UPM, 43400 Serdang, Selangor, Malaysia Available online 14 February 2006

Abstract In this study treatment of palm oil mill efuent was carried out with the intention to produce hydrogen during the anaerobic degradation process. The hydrogen generating microora was isolated from the cow dung based on pH adjustment (pH 5) coupled with heat treatment (2 h). The microora was initially tested for its hydrogen generating capability for varying fermentation pH of 4, 5, 6 and 7 while degrading palm oil mill efuent. The results showed that the biogas generation and its hydrogen content decreased in the following order of pH 5, 6, 7 and 4. Further treatment of palm oil mill efuent was carried out at an optimized fermentation pH value of 5, for varying inuent COD concentration of 5,000; 10,000; 20,000; 30,000; 40,000 and 59,300 mg/L at a hydraulic retention time of 3; 5 and 7 d, respectively. The average biogas generation was found to be 0.42 L/g COD destroyed, with a hydrogen content of 57 2% at 7d HRT. The generated biogas was free from methane. As the hydraulic retention time increased the biogas generation also increased, with a marginal increase in the hydrogen content. For example at an initial COD concentration of 59,300 mg/L for a hydraulic retention time of 3; 5 and 7 d, the hydrogen generation were found to be 52.2; 72.4 and 102.6 mL respectively. The average volatile fatty acid content in the reactor was found to be in the range 1215 130 mg/L when the inuent COD concentrations were in the range 20,00059,300 mg/L. In the case of inuent COD concentration ranging between 5,000 and 10,000 mg/L, the average volatile fatty acid was found to be 830 90 mg/L. 2006 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.
Keywords: Palm oil mill efuent; Biohydrogen; Anaerobic digestion; Cow dung; Anaerobic contact lter

1. Introduction As the reserves of oil and gas are being depleted, security of energy supply has raised the demand towards the establishment of hydrogen economy. Sustainable hydrogen energy seems to be a logical conclusion to numerous environmental problems like acid rain, green house gases and overcoming the local and transboundary pollutants [1].

Corresponding author. Tel.: +60 6 03 8946 6416; fax: +60 6 03 8946 6425. E-mail address: vijay@eng.upm.edu.my (K. Vijayaraghavan).

There are many techniques available to harness hydrogen from fossil fuel, water and biomass. Among these hydrogen generation from biomass seemed to be favored as it takes care of the degradation of the waste and yield hydrogen as a byproduct. Moreover fermentation reactions are less energy intensive and independent of light requirement [2,3]. Anaerobic treatment with the intention to generate hydrogen from wastewater and solid waste has received considerable attention during the recent years, as the generated hydrogen and its combustion product are not green house gases [4]. Various attempts have been made to generate hydrogen from wastewater like paper mill [5], municipal solid waste [6,7], starch efuent [8], food

0360-3199/$30.00 2006 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.ijhydene.2005.12.002

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processing [9], domestic waste [10,11], rice winery [12] dairy waste [13]. Numerous work had been conduced successfully towards generating hydrogen from substrate like glucose [1418], sucrose [1923]. The major source of wastewater generation from palm oil mill are namely sterilizer condensate, hydrocyclone waste and separator sludge [24]. On an average 0.91.5 m3 of palm oil mill efuent (POME) is generated for each ton of crude palm oil produced [25]. The palm oil mill efuent is rich in organic carbon with a biochemical oxygen demand (BOD) value higher than 20 g/L and nitrogen content around 0.2 and 0.5 g/L as ammonia nitrogen and total nitrogen [26]. Atif et al. [27] studied the effect of hydrogen production from palm oil mill efuent using microora isolated form the sludge of an anaerobic pond treating palm oil mill efuent. The batch experiments showed a total yield of 4708 mL H2 /L of POME with a maximum evolution rate of 454 mL H2 /L POME hr. Microora isolated for sewage [14,19,2836], tomato eld soil [3] and potatoes/soya eld soil demonstrated hydrogen generating capability [34]. So far much of the isolated microoras for hydrogen generation are derived either from sewage, soil or pure culture. A new source of microora was tested for its hydrogen generating capability namely cow dung, which showed a promising sign towards hydrogen generation using anaerobic agar as substrate. In this article the isolated microora was tested for its ability to generate hydrogen from palm oil mill efuent. The efciency of the anaerobic process was evaluated based on the amount of organic matter destroyed, biogas generated and its hydrogen content for varying inuent COD concentration, pH and hydraulic retention time (HRT).

ter was measured in terms of BOD5 . The total nitrogen was determined by Kjeldhal method, whereas the volatile fatty acid content by distillation method. The total and volatile solids were determined at 105 C and 550 50 C [37]. The hydrogen and methane content in the biogas was determined by Drager method [38]. 2.3. Preservation of wastewater The raw palm oil mill efuent was collected from the collection pit of Golden Hope Plantation, Banting, Malaysia whose characteristics are shown in Table 1. The POME was preserved at a temperature less than 4 C but above freezing in order to prevent the wastewater from undergoing biodegradation due to microbial action [37]. 2.4. Isolation of seed microora from cow dung The isolation experiments were carried out by subjecting the cow dung having a solids content of 10%, to a pH adjustment at 5 0.1 for a retention period of 3 h. Thereafter the cow dung was subjected to heat treatment at 105 C for 2 h. The microora resulting from isolation experiments were initially tested for its hydrogen generating capability for a period of 4 weeks using anaerobic agar as a substrate. During this period for every 800 mL of isolated microora, 6% anaerobic agar was added as a substrate at a ow rate of 200 mL/d. The composition of the anaerobic agar is presented in Table 2. The fermentation pH and gaseous constituent namely hydrogen and methane were analyzed in order to determine the effectiveness of the isolated microora. 2.5. Start-up

2. Methods and material 2.1. Anaerobic digester set-up The experimental set-up of the up-ow anaerobic contact lter is shown in Fig. 1. The reactor unit consists of acrylic column (100 mm ID 1200 mm height). Rigid circular porous plastic balls of 40 mm diameter served as a packing material. The openings in porous ball were 3 mm with a cross uted at every 1/4th of the ball diameter. The reactor is of complete mixed type. 2.2. Analytical process The organic strength of the wastewater was determined by COD. The biodegradability of the wastewa-

The start-up operation was carried out in two staged manner consisting of (a) seedling stage: carried out using isolated microora from cow dung based on with pH adjustment coupled with heat treatment, (b) acclimatizing stage: the microora were acclimatized with palm oil mill efuent. The anaerobic fermentation was commenced by charging 100% of the reactor volume with isolated microora form cow dung, which was supplemented with 12 g of anaerobic agar medium. The digester content was allowed to remain for a hydraulic retention time of one week. During the second week feeding were carried out using palm oil mill efuent having a COD of 24,000150 mg/L supplemented with 1% glucose and 0.5% anaerobic agar at a hydraulic retention time of 7 d. The same feed characteristics were maintained till the end of 3rd week. On the 4th

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9 8

200 mm 7 10 180 mm

11 600 mm

4 3 2

12

310 mm

13

100 mm

1. 2. 3. 4. 5. 6. 7.

Feed tank Feed pump Inlet line Sampling port Scum breaking pump Recirculation line Scum breaking line

8. Gas collection zone 9. Gas flow meter 10. Outlet line 11. Packing zone 12. Sludge accumulation zone 13. Bottom sludge wasting line

Fig. 1. Schematic diagram of upow anaerobic contact lter.

week the reactor was fed with palm oil mill efuent (24,000150 mg/L) supplemented with 0.5% of glucose. From the 5th week onwards palm oil mill efuent alone was fed into the anaerobic reactor at a COD 24,000150 mg/L till the end of 10th week. 2.6. Optimization of digestion pH The treatment of palm oil mill efuent was carried out at varying digestion pH namely 4, 5, 6 and 7 for a hydraulic retention time of 5 d. During the anaerobic

digestion process the reactor was monitored with respect to pH, volatile fatty acids (VFA), biogas generation, hydrogen content and COD removal. 2.7. Effect of hydraulic retention time on digestion efciency Based on the optimized pH value, the digester efciency was tested for varying inuent COD concentrations of 5,000; 10,000; 20,000; 30,000; 40,000 and 59,300 mg/L for a hydraulic retention time of 3,

K. Vijayaraghavan, D. Ahmad / International Journal of Hydrogen Energy 31 (2006) 1284 1291 Table 1 Characteristics of raw palm oil mill efuent (POME) Parametersa pH BOD COD Suspended solids Total nitrogen Temperature Concentration 3.5 0.1 24,710 59,300 17,260 692 84 1 Table 3 Microora viability test Fermentation period (week) 1 2 3 4 Cumulative biogas generation (L/week) 2.9 5.1 8.4 11.8

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Average hydrogen content (%) 53 54 54 56

Cumulative biogas generated (l/d)

a Except for pH and temperature all other parameters are in mg/L, temperature in ( C).

10 9 8 7 6 5 4 3 2 1 0
Biogas (l/d) Hydrogen (%)

70 60 50 40 30 20 10 0 1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69 Days of operation

Table 2 Characteristics of anaerobic agar medium Parametersa pH Casein enzymic hydrolysate Dextrose Sodium chloride Sodium thioglycollate Sodium formaldehyde sulphoxylate Methylene blue Agar
a Except for pH all other parameters are in g/L.

Concentration 5 0.2 20 10 5 2 1 0.002 20

Fig. 2. Experimental set-up of anaerobic digester.

5 and 7 d, respectively. The digester performance was evaluated with respect to biogas generation, hydrogen yield and COD reduction.

3. Results and discussion 3.1. Isolation of microora The microora was isolated from cow dung based on pH adjustment coupled with heat treatment. The reason for adopting pH adjusted coupled with heat treatment is to kill or suppress the methanogenic and nonsporulating bacteria. Lay et al. [39] stated that heat shock treatment resulted in enriching sporulating hydrogen bacteria like Clostridia. Oh et al. [40] investigation revealed that the heat treated inoculum at pH 6.2 or 7.5 resulted in higher hydrogen production when compared to inoculum which has been subjected to pH adjusted alone at 6.2. The viability of the isolated microora from cow dung was tested in an anaerobic jar having a capacity of 1 L at a fermentation pH of 5 0.1. During the fermentation period the substrate (6% anaerobic agar) addition was kept in continuous mode at a rate of 200 mL/d. The advantage in adopting continuous mode is that to over come substrate limitation. Cohen et al. [17] stated that interruption of feed could lead to sporu-

lation. Table 3 shows the cumulative biogas generation and its hydrogen content during the fermentation period of the microora viability test. The biogas generation showed a gradual rise with the fermentation period but the hydrogen content showed a marginal rise. For example during the end of 2nd and 4th week the cumulative biogas generation was 5.1 and 11.8 L/week, while its hydrogen content were 54 and 56%, respectively. In this present study microora isolated from cow dung based on pH adjustment coupled with heat treatment resulted in a biogas free from methane. Hence it can be concluded that the methanogenic and non sporulating bacteria are either killed or suppressed during isolation. Further experiments were carried out in the anaerobic contact lter using the isolated microora. 3.2. Start-up Fig. 2 shows the cumulative biogas generation and its hydrogen content during the start-up period. During the initial period of acclimatization the biogas generation showed a gradual rise till the 5th week, thereafter a drop in biogas generation was noticed from the early of 6th week to mid of 8th week. The reason for this type of behavior could be that, till the 4th week the palm oil mill efuent was supplemented with glucose and

Hydrogen content in biogas (%)

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80

Cumulative biogas generated (l)

Influent COD (mg/l)

70 COD removal (%) 60 50 40 30 20 10 0 3

10,000 59,300 HRT: 5 d

70 60 50 40 30 20 10 0 3

Influent COD (mg/l) 10,000 59,300 HRT: 5 d

5 pH

5 pH

Fig. 3. COD removal versus fermentation pH.

Fig. 4. Cumulative biogas generation versus fermentation pH.

60

thereafter its addition was stopped leading to this type of biogas generation pattern. Glucose being a simple sugar is preferred by the microora, whereas the palm oil mill efuent is a complex substance that needs sufcient time for the degradation to occur. On the mid of 8th week onwards a gradual rise in biogas generation pattern was observed. This clearly shows that the microbes are adapted to degrade the palm oil mill efuent. On the 9th and 10th week of start-up period the biogas generation were almost consistent. Hence it can be concluded that the reactor is in stable operating condition. Moreover the hydrogen content of biogas was 52 3% during the 3rd to 5th week, thereafter it ranged between 57 2% till the end of 10th week. 3.3. Effect of pH on hydrogen generation from palm oil mill efuent The effect of pH on the degradation of palm oil mill efuent for a HRT of 5 d is shown in Fig. 3. The anaerobic digestion was carried out for varying initial pH of 4, 5, 6, and 7, respectively, for an inuent COD concentration of 10,000 and 59,300 mg/L. At pH 4, irrespective of the initial COD concentration the COD removal percent was found to be lowest when compared to other operating pH namely 5, 6 and 7. A maximum COD removal occurred when the digester was operated at pH 5. The digester showed COD removal efciency in the decreasing order of pH 5, 6, 7 and 4, respectively. For an initial COD concentration of 10,000 mg/L at a digestion pH of 4, 5, 6 and 7, the COD removal efciencies were found to be 36, 67, 62 and 59%, respectively. In the case of an inuent COD concentration of 59,300 mg/L at pH 4, 5, 6 and 7, the corresponding COD removals were found to be 15, 29, 25 and 22%, respectively. The

50
Hydrogen content (%)

40 30 20 10 0 3 4 5 pH 6 7 8
Influent COD(mg/l) 10,000 59,300 HRT: 5 d

Fig. 5. Hydrogen content versus fermentation pH.

possible reason for low COD removal efciency at pH 4 could be due to the change in metabolic reaction resulting in shift in intermediate production pathway from acid production phase to solvent production phase as stated by Khanal et al. [41] and Byung and Zeikus [42]. Figs. 4 and 5 show the cumulative biogas generation and its corresponding hydrogen content during varying digestion pH of 4, 5, 6 and 7, respectively. The cumulative biogas generation and its hydrogen content varied depending on the digestion pH for a given organic strength. As shown in Fig. 4 for an inuent COD of 59,300 mg/L of palm oil mill efuent at 5 d HRT for a digestion pH of 4, 5, 6 and 7, the cumulative biogas generation were found to be 42, 73, 69 and 60 L respectively. Whereas the corresponding hydrogen contents for the above said condition were found to be 31, 56, 53 and 51%, respectively, as shown in Fig. 5. At pH 5 the biogas generation and its hydrogen content was high, while at pH 6 and 7 there was a marginal drop. Irrespective of digestion pH the biogas was free from

K. Vijayaraghavan, D. Ahmad / International Journal of Hydrogen Energy 31 (2006) 1284 1291

80 70
Cumulative biogas generated (L)
HRT (d) 3 5 7

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120 100 80 60 40 20 0 0 10000 20000 30000 40000 50000 60000 70000 Influent COD (mg/l)
HRT (d) 3 5 7

COD removal (%)

60 50 40 30 20 10 0 0

10000 20000 30000 40000 50000 60000 70000 Influent COD (mg/l)

Fig. 6. COD removal versus varying inuent COD concentration.

Fig. 7. Cumulative biogas generation versus inuent COD.

Hydrogen content in biogas (%)

methane content. Earlier research have stated that the maximum hydrogen generation occurred at a pH of 6.2 [40] and 5.56.0 [36,41]. 3.4. Treatment of palm oil mill efuent based on the optimized pH value A pH value of 5 was found to be the optimum towards hydrogen generation from the palm oil mill efuent. Based on the optimized pH value the treatment of palm oil mill efuent was carried out at varying inuent COD concentrations of 5,000; 10,000; 20,000; 30,000; 40,000 and 59,300 mg/L for different hydraulic retention time of 3, 5 and 7 d, respectively. Fig. 6 shows the COD removal percent versus inuent COD concentration for an HRT of 3, 5 and 7 d. In the case of 3, 5 and 7 d HRT, for an inuent COD concentration above 20,000 mg/L the COD removal efciency decreased with the increase in the inuent COD concentration. However, as the HRT increased the COD removal also increased. For example at an inuent COD concentration of 20,000 mg/L for a HRT of 3, 5 and 7 d, the COD removal were found to be 48, 66 and 73%, respectively. Fig. 7 shows the cumulative biogas generation versus inuent COD concentration. The biogas generation showed an increasing trend with the rise in hydraulic retention time. For example above an inuent COD concentration of 20,000 mg/L the biogas generation lowered with the increase in inuent COD concentration. Whereas as the hydraulic retention time increased the biogas generation also increased. The possible reason for low biogas generation at 3 d HRT could be due to the accumulation of intermediate products. As the HRT increased to 5 and 7 d a higher percentage of metabolic reaction could have reached the end point resulting in a

100 90 80 70 60 50 40 30 20 10 0 0 10000 20000 30000 40000 50000 60000 70000 Influent COD (mg/l)
HRT (d) 3 5 7

Fig. 8. Hydrogen content versus inuent COD.

higher gaseous end product. Fig. 8 shows the hydrogen content of the biogas for varying hydraulic retention and inuent COD concentrations. Even though the biogas generation (Fig. 7) increased with the increase in hydraulic retention time the corresponding hydrogen content (Fig. 8) showed a marginal rise. For example at an inuent COD concentration of 20,000 mg/L for a HRT of 3, 5 and 7 d, the biogas generation were found to be 45.2, 56.3 and 63.9 L, whereas the corresponding hydrogen content were 53, 55 and 56%, respectively. In the case of inuent COD concentration of 59,300 mg/L for a HRT of 3, 5 and 7 d, the biogas generation were found to be 52.2; 72.4 and 102.6 L, whereas the corresponding hydrogen content were 56, 57 and 59%, respectively. In hydrogen content in the biogas did not show any variation as the inuent COD concentration was increased. Hence it can be concluded that the metabolic reaction of the hydrogen generating bacterial species occurs in a steady phase.

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1600 Volatile fatty acids (mg/l) 1400 1200 1000 800 600 400 200 0 0

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HRT (d) 3 5 7

10000 20000 30000 40000 50000 60000 70000 Influent COD (mg/l)

Fig. 9. Volatile fatty acid content versus inuent COD.

mill efuent. Based on the optimized pH value, treatment of palm oil mill efuent was carried out using varying inuent COD concentrations namely 5000; 10,000; 20,000; 30,000; 40,000 and 59,300 mg/L for a hydraulic retention time of 3, 5 and 7 d, respectively. For the above said inuent COD concentration at a hydraulic retention time of 7 d, the COD removal efciencies were 64; 70; 73; 52; 44 and 40%, respectively. The average volatile fatty acid content in the reactor was found to be in the range of 1215 130 mg/L when the inuent COD concentration was in the range of 20,00059,300 mg/L. In the case of inuent COD concentration ranging between 5,000 and 10,000 mg/L the average volatile fatty acid was found to be 830 90 mg/L, respectively. Acknowledgements This research was supported by the Fundamental Research Grant of Universiti Putra Malaysia, Project Number: 02-03-03-057J/55180. References
[1] Maddy J, Cherryman S, Hawkes FR, Hawkes DL, Dinsdale RM, Guwy AJ, et al. Hydrogen-2003, University of Glamorgan Pontypridd, Wales, UK. [2] Das D, Veziroglu TN. Hydrogen production by biological processes: a survey of literature. Int J Hydrogen Energy 2001;26:1328. [3] J. Gorman, Hydrogen: the next generation. Science News 2002. [4] Koroneos C, Dompros A, Roumbas G, Moussiopoulos N. Life cycle assessment of hydrogen fuel production processes. Int J Hydrogen Energy 2004;29:1143450. [5] Idania VV, Richard S, Derek R, Noemi RS. Hctor M. PV Hydrogen generation via anaerobic fermentation of paper mill wastes. Bioresource Technology, Available online 18 April 2005. [6] Idania Valdez-Vazquez, Elvira Ros-Leal, Fernando EsparzaGarca, Franco Cecchi, Hctor M. Poggi-Varaldo Semicontinuous solid substrate anaerobic reactors for H2 production from organic waste: mesophilic versus thermophilic regime. Int J Hydrogen Energy 2005;30:138391. [7] Lay JJ, Lee YJ, Noike T. Feasibility of biological hydrogen production from organic fraction of municipal solid waste. Water Res 1999;33(11):257986. [8] Zhang T, Liu H, Fang HHP. Biohydrogen production from starch in wastewater under thermophilic condition. J Environ Manage 2003;69:14956. [9] Shin HS, Youn JH, Kim SH. Hydrogen production from food waste in anaerobic mesophilic and thermophilic acidogenesis. Int J Hydrogen Energy 2004;29:135563. [10] Van Ginkel S, Oh SE, Logan BE. Biohydrogen gas production from food processing and domestic wastewaters. Int J Hydrogen Energy 2005;30:153542. [11] Claassen PAM, van der Wal FJ, van Noorden DE, Elbersen HW, van Wichen JM. Bioprocess for hydrogen and methane production in Wageningen. In: 12th European conference and

The volatile fatty acid content versus inuent COD concentration for varying hydraulic retention time is shown in Fig. 9. At an inuent COD concentration ranging between 5,000 and 10,000 mg/L the VFA content ranged between 830 90 mg/L as acetate. In the case of inuent COD concentration ranging between 20,000 and 59,300 mg/L the VFA content ranged between 1215 130 mg/L as acetate. Irrespective of inuent COD concentration the volatile fatty acids showed an increasing trend as the hydraulic retention time increased. Lay et al. [39] stated that high solid organic waste such as egg, lean meat, fat meat, chicken skin, potato and rice yielded a VFA content of 18; 35; 4; 11; 23 and 5 g/L as acetate at 6.25 d HRT, when heat shock digested sludge from pig manure was used as a seeding material. Horiuchi et al. [43] stated an average volatile fatty content of 3500 mg/L during the anaerobic acidogenesis at pH 5. While Chang et al. [23] stated a VFA value ranging between 11,083 and 13,693 mg COD/L at an HRT of 4 to 24 h, while treating synthetic substrate in UASB reactor.

4. Conclusion The microora isolated from cow dung based on pH adjustment coupled with heat treatment proved to be promising candidate towards hydrogen generation. The isolation experiments were carried out by subjecting the cow dung to an initial pH of 5 for 3 h, followed by heat treatment at 105 C for 2 h. As pH play a vital role towards the metabolic reaction, optimization of pH were carried out by conducting the experiments at different fermentation pH namely 4; 5; 6 and 7, respectively. At pH 5 maximum COD removal, biogas generation and hydrogen content were observed while treating palm oil

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