Regional Annual Fundamental Science Symposium (RAFSS 2011)

Regional Annual Fundamental Science Symposium (RAFSS 2011)

http://www.ibnusina.utm.my/rafss2011

A complete isolation of cellulose, hemicellulose and lignin from palm oil trunk (POT)
Tengku Nur Zulaikha Tengku Malim Busu1 , Safiah Syazana Mohtar1 , Hanapi Mat1,2*
1 2

Advanced Materials and Process Engineering Laboratory, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, 81310 UTM Skudai, Johor, Malaysia. Novel Materials Research Group, Nanotechnology Research Alliance, Universiti Teknologi Malaysia, 81310 UTM Skudai, Johor, Malaysia

ABSTRACT Cellulose, hemicelluloses and lignin compounds were isolated from oil palm trunk by acidic organosolvent treatment followed by hydrogen peroxide bleaching and two stage precipitation. The organosolvent treatment ensures the purity of cellulose as well as recovery of hemicelluloses and lignin compounds. Organic acids were more effective than alcohols on the degradation of lignin and hemicelluloses. Formic acid/acetic acid/water (30/60/10, v/v/v) system was found to be the most effective in delignification and the removal of non-cellulose polysaccharides from the OPB, and did not have any undesirable effects on lignocellulosic compound properties such as its molecular structure and thermal stability. By using 0.1% HCl as a catalyst at 85 oC for 4 h, the treatment removed 94.1% of the original lignin and 76.5% of the original hemicelluloses. The cyanamide activated hydrogen peroxide bleaching degraded substantial amounts of residual hemicelluloses and lignin, produced the cellulose samples having a relatively high purity. Under a best condition, the isolation process yielded relatively amount (% dry weight) of cellulose, hemicellulose and lignin which are 41.37%, 15.7% and 16.43% respectively. Cellulose, hemicelluloses and lignin from both preparations were further characterized by FTIR analysis and TGA analysis. | Isolation | Oil palm biomass | Cellulose | Hemicellulose | Lignin| 2010 Ibnu Sina Institute. All rights reserved.

1.

INTRODUCTION In this study, a modified sequential treatment was adopted to extract cellulose, hemicelluloses and lignin from oil palm biomass cell wall. 2. EXPERIMENTAL

Cellulose, hemicellulose and lignin are included as polysaccharides component of plant biomass that is found in plant cell walls such as wood and cereal straws. Cellulose is a linear polymer of glucose and hemicelullose is a branched polymer of xylose whereas, lignin is a phenolic polymeric complex that attaches with cellulose and hemicelullose [1]. These materials are particularly attractive because of their relatively low cost and plentiful supply. In recent years, there has been an increasing trend towards more efficient utilization of these components in industrial applications such as food additive, fuel source and others. Recently, cellulose, hemicelluloses and lignin is being isolated from agro-industrial residues such as sugarcane bagasse [2,3], waste woods [4] and cereal straw [5]. As there is an abundant waste of oil palm biomass which are empty fruit bunch (EFB), oil palm frond (OPF) and oil palm trunk (OPT), this raw materials is highly selective. It also will be a solution in oil palm waste management in oil palm industry. These compositions have demand in industry where value added products is highly recommended [6]. Although the methods in isolation of cellulose, hemicelluloses and lignin from oil palm biomass has been investigated by some authors, the investigation only focus on the extraction of individual or two components rather on the complete isolation for all those component without affecting the quality and yield of the extracted components.
Corresponding author at: Department of Chemical Engineering, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, 81310 UTM Skudai, Johor, Malaysia E-mail addresses: hbmat@cheme.utm.my (Hanapi Mat)

2.1 Materials, method and instruments Oil palm trunk was kindly supplied by local plantation field in form of dried and small pieces. All chemicals used were of analytical and reagent grade. Infrared (IR) spectra were recorded on Shimadzu 8000 or Perkin-Elmer series 1600 spectrometers as thin film for liquid samples or KBr pellet for solid samples. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) were grafted on simultaneous thermal analyzer (NETZSCH STA-409). 2.2 Organosolvent treatment The extraction of cellulose from all the trunk was performed according to Xu et al. [5] and Sun et al.[1]. Briefly, the samples of trunk were grinded and sieved to 0.8 to 1.0mm size to make sure of efficient isolation process. Then, the grinded trunk was dried in oven at 53oC for 16 hours. Soxhlet extraction using solvent toluene:ethanol (2:1)v/v was done to remove wax. Trunk samples were weighed as solvent to trunks ratio is 10:1(ml/g). De-wax process will be run for 16 hours at maintain temperature, 110oC. The de-waxed trunk samples will be dried to remove solvent from samples in oven at 53oC. Dried and de-waxed 1

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trunk samples were soaked in two types of solvent which are formic acid/acetic acid/water (60/30/10) v/v/v with stirring at 400rpm in a 85oC water bath for 4 hours. The ratio of trunk to solvent formic acid/acetic acid/water is 1:20(g/mL). The crude cellulose was vacuum filtered using Whatman filter paper no. 4 through a Buchner funnel. The residue of filtration which is extracted crude cellulose will be rinsed with ethanol and distilled water to remove residual acid-soluble and alkali-soluble material. The rinsed crude cellulose will be dried in oven at 53oC for 16 hours whereas, the black liquor filtrates was kept for hemicellulose and lignin isolation. The organic solvent black liquor (filtrate) was left without neutralization. The black liquor then was concentrated at reduced pressure to remove the organic solvent from unstable polysaccharides using rotary evaporator. Then, the mixture was precipitated using 3 volume of 95% ethanol in centrifuge at 3000g for 30 minutes. The precipitate in the form of pellet that was obtained is identified as hemicellulose A and was further washed with 50mL of 95% (v/v) acidified ethanol at pH 2 to remove residual acid from the samples. Hemicellulose A then was air dried for 16 hours to evaporate the ethanol. The supernatant (from the first precipitation to get hemicellulose A) was preceded with re-precipitation of isolated lignin. The supernatant was evaporated to remove all the ethanol.

Then, the supernatant is mixed with 20% HCl to pH 1.5 and was kept for 24 hours at 4oC. The mixture will be centrifuged at 3000g for 15min. The precipitate that was obtained is identified as lignin A. Lignin A was further washed with 50mL acidified water at pH 2 and was freeze dried in a refrigerator. Both hemicellulose A and lignin B was kept freeze for characterization. The dried hemicelluloses A and lignin A were then collected and stored in an airtight bottle at room temperature. The residue from previous treatment was preceded by bleaching process. The hydrogen peroxide bleaching of the above residue was performed by post treatment with 1.8% H2O20.18% cyanamide (residue:extractant, 1:30, g/ml) at 50oC water bath under pH 10 for 4 h, respectively. The bleached cellulose obtained was filtered and washed with water and ethanol. Finally, cellulose was dried in an oven at 60oC for 16 h. The filtrate was used in isolation of hemicelluloses B and lignin B as a procedure used in isolation of hemicelluloses A and lignin A. Cellulose, hemicelluloses B and lignin B were freeze dried for characterization and then collected and stored in an airtight bottle at room temperature. Yield of cellulose, hemicellulose and lignin is given on a dry weight basis related to the starting material.

Figure 1. Isolation of cellulose, hemicellulose and lignin from oil palm biomass

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2.3 Characterization of cellulose, hemicellulose and lignin preparation FTIR spectrum analysis

sample weighed between 8 and 12 mg. Each sample was heated from room temperature to 600oC at a rate of 10oC per minute [2]. 1. RESULTS & DISCUSSION Yield and chemical composition

The infrared spectra of the cellulose, hemicelullose and lignin will be obtained on an FTIR spectrophotometer model Perkin Elmer System 2000 spectrophotometer in the frequency range of 4000-400 cm-1 using a KBr disc. The pressed-disk technique was employed for disk preparation, where 950mg of dried KBr and 5mg of sample were used. The KBr and sample mixtures were intimately mixed using pestle and mortar before being placed in a special die and compressed into a small disk. After the disk had been removed from the die, it was placed in a suitable holder in the spectrophotometer and scanned directly [6]. Thermal Analysis Thermal analysis of cellulose, hemicellulose and lignin preparations was performed using thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) on a simultaneous thermal analyzer (NETZSCH STA-409). The apparatus was continuously flushed with nitrogen. The

3.1

The yield of cellulose, hemicelluloses and lignin isolated from oil palm trunk are given in Table 1. The results showed that the bleaching of crude cellulose residues with 1.8% H2O20.18% cyanamide at 50 oC under pH 10.for 4 h released C1, 41.37wt% (% dry starting material) of cellulose. Then, precipitation of organic acid black liquor and bleaching filtrate released H1, 15.7 wt% of hemicelluloses and L1, 16.43 wt% of lignin. These results showed that the treatment with formic acidacetic acidH2O (30/60/10, v/v/v) using 0.1% HCl as a catalyst at 80 oC for 4h gave relatively high yields of cellulose, hemicelluloses and lignin. This high solubility of lignin than hemicelluloses indicate that lignin is dissolved or degraded more easily in the organosolv treatment as delignification occurred effectively during the organosolv treatment.

Table 1. Lignocellulosic materials yield of Oil Palm Trunk (%, Dry Weight). Lignocellulosic materials Lignin, L1 a Hemicellulose, H1 Cellulose, C1 Composition (wt%) 16.43 15.70 41.37

It has been known for a long time that acetic acid acts as a solvent for lignin [8]. In this case, the acid catalyses hydrolysis of lignin and hemicelluloses, and its solvent dissolves the lignin fragments. In addition, a high concentration of acetic acid and formic acid is necessary in the treating liquor since the pH in solution need to be low enough to accelerate lignin hydrolysis as for high removal of lignin in pulping process [1]. Recently, chlorine-free bleaching process has been concerned as environmental issue. Thus, this alkaline peroxide is one of the solutions [9]. Alkaline peroxide chemical bleaching involves mainly electrophilic reactions of hydrogen peroxide as delignifying agents. The release of hemicelluloses and lignin during the alkaline peroxide bleaching process was suggested to be due to direct attack on a-aryl ether bonds between lignin and hemicelluloses. Although they are strong bleaching reagents, they are, however, limited in their effectiveness in degrading lignin [1].In order to increase the effectiveness without cause degradation of polysaccharide, the activation of oxygenbased reagents was being extensively studied. One particular area of interest has been in the activation of 3

hydrogen peroxide by cyanamide (H2NCN) [1,10,11,12]. This activator improves peroxide bleaching by increased lignin removal and results in an increased brightness [1,12,13]. Therefore, the present study has also been extended to the bleaching of the organosolv treated wheat straw with cyanamide activated hydrogen peroxide under alkaline conditions. 3.2 FTIR Spectra

Among the analysis techniques described in the literature, FT-IR spectroscopy shows interesting characteristics such as high sensitivity and selectivity, high signal-to-noise ratio, accuracy, data handling facility, mechanical simplicity and short time and small amount of sample required for the analysis [14]. In addition, the spectrum of a cellulose, hemicelluloses and lignin sample gives an overall view of its chemical structure. Figure 2 shows the FT-IR spectra of the cellulose sample C1. The major features of the spectra are the occurrence of three ester bands at 1382 (CCH3), and C O stretching band at 1269 cm-1 due to partial acetylation of

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hydroxyl groups in both polysaccharides and residual lignins [15], indicating that the cellulose was acetylated to a noticeable degree during the treatment with acetic acid/water or formic acid/acetic acid/water system The absorption at 3407 cm-1 is assigned to stretching of -OH groups and that one at 2923 cm-1 to the CH stretching. The band at 1634 cm-1 relates to the bending mode of the absorbed water. A noticeable peak at 1456 cm-1 is due to the CH2 bending. The absorbance at 1321 cm-1 originated from the CC and CO skeletal vibrations [4]. The peak at 1159 cm-1 arises from CO anti-symmetric bridge

stretching. The absorption band at 1106 cm-1 is attributed to COH skeletal vibration. The COC pyranose ring skeletal vibration gives a prominent band at 1034 cm-1. A small sharp band at 897 cm-1 represents the glycosidic C1H deformation with ring vibration contribution and OH bending, which is characteristic of b-glycosidic linkages between glucose in cellulose [16]. The spectrum do not show the carbonyl band at ~1739 cm-1(C=O) due to the saponification. The absence of the band at ~1513 cm-1 in the spectra indicates that the cellulose samples are relatively free of residual lignin [1].

Figure 2. Fourier transform infrared (FT-IR) spectra of cellulose extracted with organosolvent treatment at 80oC for 4 h from oil palm trunk. Figure 3 shows FT-IR spectra of lignin sample isolated from OPB trunk, L1. The spectral profiles and the relative intensities of the bands were rather similar in all six spectras, which confirmed that the main of lignin structure did not change significantly during the organic acid treatment Aromatic skeleton vibrations occur at 1513 and 1456 cm1, in which the aromatic semicircle vibration (a vibration involving both C-C stretching and a change of the HCC bond angle) [17]. The weak band at 1700 cm1 is originated from the carbonyl and unconjugated ketone and carboxyl group stretching revealed that a noticeable oxidation of the lignin structure did occur during the organic acid treatment. Moreover, aliphatic CH stretchs in CH3 and syringyl ring breathing with CO stretching are clearly seen at 1382 and 1321 cm1, respectively. Furthermore, the intensity of band at 1374 cm1 was really low indicating that the oxidation occurred mainly at - position of lignin side chains during the organic acid treatment. The bands at 1112 and 1040 cm1 are indicative of the aromatic CH in-plan deformation for syringyl type and guaiacyl type, respectively. Aromatic CH out of bending exhibits at 844 cm1[5].

Figure 3. Fourier transform infrared (FT-IR) spectra of lignin extracted with organosolvent treatment at 80oC for 4 h from oil palm trunk. Figure 4 shows the FT-IR spectra of hemicelluloses, H1 obtained by precipitation in 3 volumes of 95% ethanol solutions from acid organic black liquor respectively. The absorption at 3434 cm-1 is attributed to the stretching of -OH groups. The C-H stretching vibration gives signals at 2935 and 2881 cm-1. The band at 1638 cm-1 is due to the bending mode of absorbed water. Obviously, the hemicelluloses samples showed the typical signal pattern for the hemicelluloses and had a specific band in the 1300-1000 cm-1 region, which is dominated by ring vibrations overlapped with stretching vibrations of side 4 groups (C-OH) and the glycosidic bond vibration (C-OC). The high absorbance at 1333 cm-1 arises from the C-C and C-O skeletal vibrations. In the anomeric region (950700 cm-1), a small band at 905 cm-1, which is due to the C-1 group frequency or ring frequency, is indicative of glycosidic linkages in hemicelluloses, whereas small peaks at 778 cm-1 in spectrum is characteristic of Ranomers in side chains. The small bands at 1462, 1423, 1333, 1276,and 1235 cm-1 represent C-H and C-O or OH bending vibrations in hemicelluloses, respectively. Evidently, the occurrence of an intensive band at 1513

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cm-1 in spectrum is due to aromatic skeletal vibrations in presence of small amount of bound lignin, in the

precipitated hemicellulose obtained in 3 volume of 95% ethanol solution [2].

Figure 4. Fourier Transform Infrared (FT-IR) spectra of hemicellulose extracted with organosolvent treatment at 80oC for 4 h from oil palm trunk.

(a)

(b)

(c) Figure 5. Thermogravimetric analysis/differential scanning calorimetry (TGA/DSC) curves of the (a)cellulose, (b)hemicelluloses and (c)lignin of oil palm trunk. 3.4 Thermal Stability thermogravimetric method. Figure 5.0 gives the TGA curves for cellulose (a), hemicelluloses (b) and lignin (c) respectively. TG curves indicate the weight loss of lignin samples in relation to the temperature of thermal 5

Thermal stability of the cellulose, hemicelluloses and lignin of oil palm trunk was investigated by

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degradation. As can be seen, all samples began to decompose at 250oC (C1), 200oC (H1) and 190oC (L1). When weight loss reached 50%, the temperature increased to 350oC (C1), 870oC (H1) and 880oC (L1). The thermal degradation of the cellulose sample took place rapidly when it started to decompose but hemicelluloses and lignin sample degraded gradually. The differences between these thermal stability corresponded to their significant difference of molecular weights. 4. CONCLUSION The treatment of oil palm trunk with combination of organic acids and alkaline hydrogen peroxide treatment is very well suitable for separation of the principal lignocellulosic components. This is practically true for isolation of celluloses, hemicelluloses and lignins. FTIR spectra indicated that the celluloses isolated by treatment with aqueous organic acid and hydrogen peroxide bleaching gave typical spectra. In addition, remarkable spectra of hemicelluloses and lignins suggested that organic acid treatment and precipitation of its black liquor did not give significant degradation of their molecule. Thermal stability also shows acceptable value of thermal degradation.

Research Program (Project No. 05-01-06-SF1006), Research University Grant (GUP 00H63) from UTM and MOHE, and the 2009 ExxonMobil Research Grant are gratefully acknowledged. REFERENCES
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ACKNOWLEDGEMENT The financial supports from the Ministry of Agriculture (MOA), Malaysia under the eScience

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