EXTRACTION OF CELLULOSE FROM OIL PALM EMPTY FRUIT BUNCH

(OPEFB) AND PREPARATION OF TRANSPARENT CELLULOSE THIN FILM

NUR SYAZWANA BINTI IZZAHA

Malaysian Institute of Chemical and Bioengineering Technology

Universiti Kuala Lumpur

JANUARY 2020
EXTRACTION OF CELLULOSE OIL PALM EMPTY FRUIT BUNCH (OPEFB) AND
PREPARATION OF TRANPARENT CELLULOSE THIN FILM

NUR SYAZWANA BINTI IZZAHA

55217117071

A thesis submitted in fulfilment of the

requirements for the award of degree of
Bachelor of Chemical Engineering Technology (Hons.) in Bioprocess

Malaysian Institute of Chemical and Bioengineering Technology

Universiti Kuala Lumpur

JANUARY 2020
 TABLE OF CONTENT

LIST OF TABLES ..................................................................................................... v

LIST OF FIGURES ....................................................................................................vi

CHAPTER 1: INTRODUCTION ................................................................................. 1

1.1 Research background ...................................................................................... 2

1.2 Problem statement ........................................................................................... 3

1.3 Research objectives ......................................................................................... 4

1.4 Significant of the study ..................................................................................... 4

CHAPTER 2: LITERATURE RIVIEW ........................................................................ 6

2.1 Palm Oil Plantation in Malaysia ........................................................................ 6

2.2 Waste generated from Oil Palm Industry .......................................................... 9

2.3 Cellulose ........................................................................................................ 11

2.4 Nanocellulose ................................................................................................ 13

2.5 Transparent Nanocellulose Thin Film ............................................................. 15

CHAPTER 3: METHODOLOGY .............................................................................. 17

3.1 Apparatus, chemicals and materials used for the experiment . Error! Bookmark
not defined.

3.1.1 Collection of Samples.......................................... Error! Bookmark not defined.

3.1.2 Chemicals used .................................................... Error! Bookmark not defined.

3.1.3 List of apparatus used ......................................... Error! Bookmark not defined.

3.2 Preparation of sample ....................................... Error! Bookmark not defined.

3.3 Experimental procedure .................................... Error! Bookmark not defined.

 3.3.1 Extraction of OPEFB – Cellulose using Soda Pulping Process with
parameters manipulation .................................... Error! Bookmark not defined.

3.3.2 Alkali Bleaching Treatment ................................. Error! Bookmark not defined.

3.3.3 Nanocellulose preparation using Acid Hydrolysis Treatment ............... Error!

Bookmark not defined.

3.3.4 Preparation of Nanocellulose Solution ............. Error! Bookmark not defined.

3.3.5 Preparation of Nanocellulose Film .................... Error! Bookmark not defined.

3.4 Analysis ............................................................ Error! Bookmark not defined.

3.4.1 .Scanning Electron Microscope (SEM) ............. Error! Bookmark not defined.

3.4.2 Transmission Electron Microscope (TEM) ....... Error! Bookmark not defined.

3.4.3 Fourier Transmission Infrared Spectroscopy (FTIR) ..... Error! Bookmark not
defined.

3.4.4 X-Ray Diffraction (XRD) ...................................... Error! Bookmark not defined.

3.4.5 Thermogravimetric Analysis (TGA) ................... Error! Bookmark not defined.

3.5 Methodology Flowchart ..................................... Error! Bookmark not defined.

CHAPTER 4: EXPECTED RESULT ........................................................................ 25

4.1 Expected Result ............................................................................................. 25

4.2 Research Costing .......................................................................................... 27

4.3 Research Planning ......................................................................................... 28

4.3.1 Gantt Chart......................................................................................................... 29

REFERENCES ........................................................................................................ 31
 LIST OF TABLES

TABLE TITLE PAGE

2.1 The chemical composition of oil palm biomass waste 12

3.1 The ranges of independent variables for cellulose 19

extraction process

4.1 Typical functional group of cellulose absorption peaks 26

4.2 The cost for each materials per unit mass or volume (kg/L) 27

4.3 Final Year Project Gantt Chart 29

v
 LIST OF FIGURES

FIGURE TITLE PAGE

2.1 The oil palm trees 6

2.2 The total planted area of palm oil in Malaysia from 1975 to 7
2011, Million hectares
2.3 The oil palm trees and the various waste form 8

2.4 The overall palm oil wastes generations 9

2.5 The structure of cellulose 11

2.6 Acid hydrolysis of cellulose reaction 13

vi
 LIST OF ABBREVIATIONS

OPEFB Oil Palm Empty Fruit Bunch

GDP Gross Domestic Product
EFB Empty Fruit Bunch
OPF Oil Palm Fronds
OPT Oil Palm Trunks
PPF Palm Pressed Fibres
TGA Thermogravimetry Analyzer
FTIR Fourier-Transform Infra-Red
PSA Particle Size Analyzer
MPOB Malaysian Palm Oil Board

1
 CHAPTER 1

INTRODUCTION

1.1 Research background

In Malaysia, agriculture is one of the most important industry. Malaysia’s

Gross Domestic Product (GDP) in agriculture sector is about 7.3 percent in 2018; with
major contributor is the oil palm sector with 37.9 percent (Hasan, 2019). Oil palm
industries produced 90 million tons of waste biomass with the largest contributor of 30
million tons from Empty Fruit Bunch. The biomass consists of lignocellulosic
component including from empty fruit bunch (EFB), oil palm fronds (OPF), oil palm
trunks (OPT) and palm pressed fibres (PPF). Cellulose can be obtained from oil palm
biomass as it is part of the lignocellulosic component. The highest cellulose content in
the biomass is 43-65% which is from oil palm empty fruit bunch (Dungani et al., 2018).

Cellulose (C6H10O5)n is natural linear polysaccharide. It is one of the most

abundant biomaterials on earth that can be synthesised by plants or some bacteria.
Plant cell walls are mainly composed by cellulose which help the plant to remain strong
and rigid whereas the other component composed together with the cellulose(Bailey,
2019). Cellulose in plant fiber are in amorphous phase that connected with crystalline
phase through intermolecular and intramolecular hydrogen bonds. The unique
properties of the cellulose are low density, good mechanical properties and
biodegradable (Wulandari et al., 2016).

2
 Cellulose in nanometer ranged is called nanocellulose. The diameter of the
particles are in the range of 10-20 nm (George & Sabapathi, 2015). Nanocellulose can
be produced using various method. It can be produced either by acid hydrolysis,
enzymatic hydrolysis or ultrasonic technique. The nanocellulose obtained by using
acid hydrolysis method has been selected by many researchers as the method gives
the smallest size particles compared to the other method. The nanocellulose has been
used in various field such as food wrappers which replacing the non-biodegradable
plastics, nanocellulose-based film and as the barrier in the separation process of
hazardous waste (Wulandari et al., 2016).

The nanocellulose-based film has several characteristics such as light in

weight, renewable, recyclable and compatible to the health and the environment. The
film can be prepared using various process including spraying, spin-coating, dip-
coating, chemical vapour deposition (CVD), evaporation and sputtering (Herrera
Rodriguez, 2015). The casting technique are widely used as the thickness of the film
produced will be distributed uniformly, maximum optical purity and low haze.
Nowadays, the solvent cast technology are used widely in productions of film as it
gives extremely high quality (Siemann, 2005).

1.2 Problem statement

In this thesis, for cellulose extraction from Oil Palm Empty Fruit Bunch
(OPEFB), the extraction using soda-pulping process has been conducted by several
researchers but the optimum parameters of the process have not yet been determined.
Thus, the parameter used in the research need to be determined carefully as it will
affect the whole extraction process. For the preparation of transparent nanocellulose
thin film, there are research that has been reported on the preparation of the film using
casting method but the film prepared using nanocellulose synthesis of cellulose
extracted from OPEFB is limited. This make the research quite challenging due to lack
of references about the information needed as the literature reported does not
focussing on the specified component. Therefore, this study will mainly focus on
parameters manipulation on cellulose extraction process and production of transparent

3
nanocellulose thin film from OPEFB as well as the nanocellulose synthesis and
preparation of transparent nanocellulose thin film.

1.3 Research objectives

This research is focusing on parameters manipulation of cellulose extraction

from OPEFB and to produce OPEFB based transparent cellulose thin film. Thus the
objective of this study are:

i. To extract cellulose using soda-pulping process by manipulating parameters

such as concentration of caustic soda used, extraction time and temperature.
ii. To prepare transparent cellulose film using casting method.
iii. To characterize the cellulose and transparent cellulose film produced.

1.4 Significant of the study

Currently, the natural fibers such as lignocellulosic biomass from oil palm
waste such as cellulose is one of the major applications in food packaging and film
industry. The cellulose obtains from OPEFB via extraction method such as soda-
pulping is widely explored and well documented in the open literature. However, the
optimization of parameters in the extraction process has not been determine. The
optimum parameters need to be determine to achieve the most optimize and effective
extraction process. Besides, the preparation of transparent cellulose thin film are also
widely explored and documented but the production of transparent cellulose thin film
from cellulose extracted from OPEFB is limited.

The analytical, thermal and morphological test will be conducted to determine

the characterization of each sample. The test will be performed by using Fourier-
Transform Infrared (FTIR) Spectroscopy, Particle Size Analyzer (PSA), Tensile and
Thermogravimetric Analysis (TGA) to determine the morphology, nanostructure,

4
functional group, crystallinity index and thermal stability of the sample respectively. By
conducting this study, the problem stated are expected to be solved and the optimum
temperature can be finalised to the extraction process.

5
 CHAPTER 2

LITERATURE REVIEW

2.1 Palm Oil Plantation in Malaysia

Elaeis guineensis jacq.or commonly known as oil palm are originally from
West Africa. The trees grow in the wild and later was developed into an agriculture
crop. It was introduced to Malaya which now known as Malaysia, in early 1870’s by
the British as an ornamental plant. The first commercial planting in 1917 took place in
Tennamaran Estate in Selangor where the foundations for the vast oil palm plantations
and the palm oil industry in Malaysia lays nowadays. The planted palm oil is the tenera
variety which is a hybrid between dura and pisifera. Palm trees can group up to sixty
feet in height or more. The young and mature plant trunks are wrapped in the fronds
which give them a rough appearance whereas apart from the scars left by the fronds
that are withered and fallen off have a smoothers trunk is known as the older trees. It
has an economic life about 25 years. After 30 months field planting, the palm oil could
begin to be harvested (“The Oil Palm Tree,” 2020).

6
 Figure 2.1 The oil palm trees

Oil palm is the main corps in Malaysia and it is widely planted all over the
country especially in Johor, Sarawak and Sabah. Malaysia has the second largest oil
palm plantation area in the world which make it the second largest producer of palm
oil and a major exporter. Oil palm is one of the most important sector that change the
agriculture and economy scenario (Abdullah & Sulaiman, 2013). The planted area in
Malaysia continuously increasing at a rapid pace since 1960. Based on figure 2, 1.5
million hectares were planted with palm tree in 1985, then the plantation increased to
4.3 million hectares in 2007 and as of 2011, the total planted area was 4.9 million
hectares (“Palm Oil Industry,” 2015). In 2018, the total planted area were stood at 5.8
million hectares (Tan & Ho, 2019).

7
 Figure 2.2 The total planted area of palm oil in Malaysia from 1975 to 2011, Million
hectares (“Palm Oil Industry,” 2015)

Part of the total planted area, 4.49 million hectares of the land in Malaysia is
under oil palm cultivation, where 17.73 million tonnes of palm oil and 2.13 tonnes of
palm kernel is produced (“The Oil Palm Tree,” 2020). The oil palm known as the most
efficient oil-bearing crop in the world as it only required 0.26 hectares of land to
produce one tonne of while compared to soybean, sunflower and rapeseed that
required 2.22, 2 and 1.52 hectares respectively, to produce the same amount of oil.
Malaysia accounted for 11% of the world’s oils and fat production and 27% of export
trade of oils and fats (“The Oil Palm Tree,” 2020).

The increase in palm oil cultivation cause abundance of raw materials

available on the palm which consisting of 90% biomass wastes and only around 10%
of the oil (Abdullah & Sulaiman, 2013). The huge amount of waste generated have led
to the concerns over the management of the wastes. The disposal method that
commonly used for agricultural waste are including mulching or leaving the waste to
decompose naturally in the plantation area. This practices can cause harmful to the
environment despite the benefit obtain.

8
2.2 Waste generated from Oil Palm Industry

Oil palm industry has been producing large amount of biomass wastes in the
field and the oil pam mills. The pressed fruit fibres (PFF), empty fruit bunch (EFB), oil
palm shell (OPS) and oil palm mill effluent (POME) are waste generated from the mill
whereas the oil palm trunk (OPT) and oil palm fronds (OPF) are comprises from the
plantation. The fronds waste are produced during pruning and harvesting while the
trunks waste produced during the replanting. Generally, each oil palm trees will gives
24% of OPF during harvesting at fresh fruit bunches in a year. Figure 3 shows the oil
palm trees and the various waste from together with its respective derivatives.

Figure 2.3 The oil palm trees and the various waste form

In 1998, about 90 million tons of waste from oil palm fruit production were
generated. EFB, OPS and PFF contributing about 43-45% of the total waste (Abdullah
& Sulaiman, 2013). Figure 4 shows the summarization of the wastes generation of the
palm oil industry in general. The largest amount of solid biomass generated for
renewable resources is from oil palm empty fruit bunch (EFB) which is 15 million
tonnes, then followed by oil palm fronds (OPF) with 13 million tonnes of biomass.

9
 Figure 2.4 The overall palm oil wastes generation (Hassan & Abd-Aziz, 2012)

The moisture content in EFB are usually dried until it reaches about 40%.
MPOB researchers has done a study on converting EFB into paper-making pulp. This
is because EFB can be categorised as fibrous crop residues that known as
lignocellulosic residues or biomass. Lignocellulosic biomass is defined as an abundant
and renewable resources that can be obtain from plants that mainly composed of
polysaccharides such as cellulose and hemicellulose and an aromatic polymer such
as lignin (Zoghlami & Paës, 2019). Besides EFB, other waste such as OPT, OPF, PFF
and OPS are also categorised as fibrous crop residues.

10
2.3 Introduction to Cellulose

The most abundant renewable in the world is the cellulose. Nearly 100 billion
tons of cellulose are estimated to be produced every year with an annual growth of
about 40 billion tons (Kihlman, Wallberg, Stigsson, & Germgård, 2011). However, in
processing only used a tiny fraction from the total amount produced. Natural plant fiber
are composed by cellulose, small number of extractives and amorphous polymer such
as hemicellulose and lignin. In 1980s, cellulose was first isolated and recognized as a
chemical substance by an agricultural chemist. Cellulose is a linear polymer that linked
by ß-1,4-glycosidic bonds. The arrangement of the isomers is in a parallel row whereas
the hydroxyl groups are arranged in adjacent chains that held together by forming the
hydrogen bonds to hydrolysis the starch. The cellulose has strong tendency to form
intramolecular and intermolecular hydrogen bonds as the molecules is arranged in
linear form (Norrihan, 2008).

In plant, cellulose is the main substances that make up the cell walls. The
cellulose is naturally hydrophilic polymer that contributes to the physical stability of the
cells. According to Bhatnagar and Sain, (2005) the primary cell wall of plant consists
of 9-25% of cellulose microfibrils, 25-50% matrix of hemicellulose and 10-35% of
pectins whereas the secondary cell wall are formed from the lignin of primary cell walls
thickening and inclusion into the cell wall matrix. The secondary cell walls contain 40-
80% cellulose, 10-40% hemicellulose and 5-25% lignin (Bhatnagar & Sain, 2005). The
cellulose is the most abundant organic compound. It does not soluble in water due to
the severe network of intermolecular and intramolecular hydrogen bonding between
the glucose unit and hydrophobic interactions between the cellulose molecules
(Medronho, Romano, Miguel, Stigsson, & Lindman, 2012). It also contains high
crystallinity and high molecular weight. The most part of the cellulose are in crystalline
structure that have a degree of polymerisation (DP) of up to 15000 (Kihlman et al.,
2011).

11
 Figure 2.5 The structure of cellulose

Figure 2.5 shows the structure of the cellulose. Natural cellulose can be found
in the form of microfibrils that organized in the fibers, cell walls and other. The cellulose
chains in cellulose microfibrils are aligned parallel to the microfibrils axis. Whereas, the
cellulose chains in the cellulose fibres are organised and oriented in ultrastructural that
responsible to their mechanical strength (Malainine et al., 2002). Cellulose has several
characteristics such as odourless, flavorless, hydrophilic, insoluble in water and it is
biodegradable material. The melting point of cellulose is 467 °C and it can be degraded
into glucose by acid treatment at high temperature. Cellulose are commercially used
for paper manufacturing, textile industry, drug fillers and as a common household
materials such as coffee fillers, sponges, laxatives and films (Helmenstine, 2019).

Studied had been reported that the cellulose is polymorphic with six different
forms which are cellulose I, II, IIII, IIIII, IVI and IVII Cellulose I is the native cellulose that
has parallel chain orientation while cellulose II is in an anti-parallel chain orientation.
Cellulose I can be converted to cellulose II by treated with alkali that is known as
Mercerisation process (Norrihan, 2008). Cellulose III is a cellulose that mercerized with
ammonia and it is similar to cellulose II but differ by the chain. Cellulose III chain is
similar to cellulose Iα and cellulose Iß that is arranged in parallel. Mercerisation process
is an old method used for cellulose fibers modification. In 1844, John Mercer of Great
Harwood in Lancashire, England, is the first to device the process by treating cotton
fibre with sodium hydroxide. The reinforcement by alkaline treatment is the main
mechanism due to the degradation of hemicellulose and other amorphous content, as
the treatment will produce more effective products than the polar extractive treatment
(Dai & Fan, 2014).

12
 Based on the previous study, oil palm empty fruit bunch has the highest
cellulose composition compared to the other fibres such as oil palm fronds, oil palm
trunk and palm pressed fibers. The solid waste that has higher potential for commercial
exploitation is EFB and OPT compared to the other types of biomass (Dungani et al.,
2018). Cellulose can be obtain from the oil palm biomass as it is part of the
lignocellulosic component. Cellulose has a diameter of 2-20 nm and 100-40000 nm
long (“Cellulose Structure,” 2018). The table below shows the chemical composition of
different oil palm fibers from the biomass that consists of lignocellulosic such as
cellulose, hemicellulose and lignin content from different studies.

Table 2.1 The chemical composition of oil palm biomass waste (Dungani et al., 2018)

Fibers Cellulose, % Hemicellulose, % Lignin, %

EFB 43-65 17-33 13-37

OPF 40-50 34-38 20-21

OPT 29-37 12-17 18-23

OPS 27-35 15-19 48-55

In the previous study, Rasli et al., (2017) stated that the extracted cellulose
treated by alkali and undergo bleaching treatment that undergo analysis using SEM
and TEM will shows a smoother surface morphology compared to the raw fibers. The
treatment will also alter the colours of the fibers from brownish to white. The changes
for both colour and morphology to become lighter and smoother is due to the removal
of hemicellulose, lignin and other extractives from the treatment and process.

2.4 Nanocellulose

Oil palm fibres consists of lignocellulosic component that are potentially

exploited in nanotechnology. Cellulose nanocrystals or nanocellulose are a
nanomaterials that are derived from the natural polymer that exist abundantly and

13
almost inexhaustible materials. The nanocellulose can be obtained primarily from
natural occurring cellulose fibers that are biodegradable. The fibers is a renewable
materials in nature, hence it is served as a sustainable and environmentally friendly
materials for applications. They are hydrophilic in nature, but they can be functionalized
to fulfil the various requirement needed. The example of the requirements are
development of high-performance nanocomposites that are using hydrophobic
polymer matrices. Due to their mechanical, optical, chemical and rheological properties,
the nanomaterials have received several significant interest (George & Sabapathi,
2015).

There are several methods that can be used to produce nanocellulose such
as acid hydrolysis, enzymatic hydrolysis or ultrasonic technique. The most common
technique used to produce nanocellulose is acid hydrolysis. This is because the
technique gives the smallest particles size among all. The hydrolysed cellulose is in
nanometer range with the particles diameter of 10-20 nm and 100-300 nm in length
(George & Sabapathi, 2015). The isolation of nanocellulose from cellulose to
nanocellulose using strong acid is known as acid hydrolysis process. Acid hydrolysis
under controlled conditions will allows the removal of the amorphous regions of
cellulose fibers while keeping the crystalline structure intact in the form of crystalline
nanoparticles. The isolated nanocellulose has been used as reinforcements in the field
of nanocomposites due to the attractive considerable attention (Sheltami, Abdullah,
Ahmad, Dufresne, & Kargarzadeh, 2012). The figure below shows the cellulose treated
using sulphuric acid to form nanocellulose by acid hydrolysis treatment.

H₂SO₄

Figure 2.6 Acid hydrolysis of cellulose reaction

14
 The nanocellulose has been used in various field such as food wrappers
which replacing the non-biodegradable plastics, nanocellulose-based film and as the
barrier in the separation process of hazardous waste (Wulandari et al., 2016). Besides
applications in food packaging, nanocomposites based on polymer matrix also has a
great potential to be applied in several fields such as aerospace, automotive,
biomedical industry and many more. The nanoscale size features with high aspect ratio
possessed excellent mechanical properties compared to composites reinforced with
micro-sized fillers. The nanoscale fibers or cellulose also gives outstanding
combination of optical, electrical, thermal, magnetic and other physical and chemical
properties (Ngadi, N. Lani, N.S. , Johari, A. , & Jusoh, 2014)

2.5 Transparent Nanocellulose Thin Film

Cellulose is an interest raw materials that attracted researchers over the last
few decades due to the abundant resources and having a unique structure with an
extraordinary renewable and biodegradable materials. The fibers have been applicable
in film, coating and the matrix of control-release systems mainly in the food packaging
area. The nanocellulose that are derived from cellulose has been used to produce a
nanocellulose-based film. The film has several characteristic such as light in weight,
renewable, recyclable and compatible to the health and the environment.

Usually, most of the film will be prepared using cellulose II structure which
then being synthesis using one of the common method such as hydrolysis to produce
nanocellulose (Zhang, Azuma, & Uyama, 2015). The solution used can be prepared
using common solvent such as N-dimethyacetamide (DMAc) or lithium chloride (LiCl)
to dissolve the nanocellulose. Not all solvents are able to dissolve the fibers. According
to Zhao et al., (2014) the nanocomposite films produced using DMAc or LiCl as the
solvent will showed good optical transparency and remarkable mechanical properties.
Besides, Zailuddin & Husseinsyah, (2016) also reported that the preparation of
cellulose regenerated using DMAc or LiCl will enhance the tensile strength and the
modulus of elasticity of the biocomposites film due to the presence of stiff and strong
cellulose from the regeneration process. The ionic liquid of DMAc or LiCl act as non-
derivatizing single-compund solvent for cellulose, thus it was found to be very effective.

15
 The film can be prepared using various process including spraying, spin-
coating, dip-coating, chemical vapour deposition (CVD), evaporation and sputtering
(Herrera Rodriguez, 2015). The casting technique are widely used as the thickness of
the film produced will be distributed uniformly, maximum optical purity and low haze.
Nowadays, the solvent cast technology are used widely in productions of film as it
gives extremely high quality (Siemann, 2005).

The previous study by Zhao et al., (2014) reported that the diameter of the
film produced is in the range of 15-40 nm. The study also characterized the morphology,
structure and the properties of the nanocomposites film using scanning electron
microscope (SEM), X-ray diffraction (XRD), thermal gravimetric analysis (TGA) and
mechanical testing. From the result obtained, it is clearly stated that the film exhibits
good optical transparency, thermal stability and remarkable enhance mechanical
properties. Based on the work provided, the cellulose nanocomposites film
manufactured gives a high performance and environmental-friendly product.

16
 CHAPTER 3

MATERIALS AND METHODOLOGY

3.1 Introduction

This chapter describes on all the chemicals employed, material used,

preparation of sample, the experimental set-up and procedure for extraction and
transparent cellulose thin film process and several related characterization conducted
to determine the properties of product that suit application proposed. The parameters
that need to be considered during the extraction process is the temperature of each
samples in this research.

3.2 Apparatus, materials and chemicals used in the research

3.2.1 Materials and chemicals

Table 3.1 list all the materials and chemicals involved in this research. 500
micron meter of OPEFB from MPOB in Selangor, Ethanol and Hydrogen Peroxide from
Sigma Aldrich, Sodium hydroxide, Dimethylacetamide and Lithium Chloride from H &
M Chemicals Company is obtained from UniKL MICET.

17
 Table 3.2: List of materials and chemicals used in the research

Raw materials/ Molecular

Application Supplier
Chemicals formula

Raw material for

Oil Palm Empty Fruit MPOB,
- cellulose
Bunch (OPEFB) Selangor
extraction

Fibre Sigma
Ethanol C2H5OH
preparation Aldrich
Bleaching
Hydrogen Peroxide H2O2 Sigma Aldrich
process
Extraction H&M
Sodium Hydroxide NaOH
process Chemicals
Dimethylacetamide
C4H9NO Film preparation H&M
(DMAc) Chemicals

Lithium Cloride LiCl Film preparation H&M

Chemicals

3.2.2 Apparatus

Table 3.2 list all the machine and apparatus used in this research for sample
preparation, cellulose extraction and transparent thin film preparation.

18
 Table 3.3: List of machine and apparatus used in the research

Machine / Apparatus Model Function / Testing

Soxhlet Extractor FAVORIT To de-wax the fibre for sample

preparation

Buchner funnel vacuum To filter the sample

pump

Orbital shaker To shake the sample in circular

motion with slow speed

Magnetic stirrer with hot To stir the sample while

plate heating

Waterbath To incubate samples in water

at constant temperature

Conventional oven To dry sample under constant

heat

Analytical balance To weight sample

Glass slide To cast the film

Fourier-Transform Infra- To identify the functional group

Red Spectroscopy of the sample
(FTIR)
Thermogravimetric To observe the thermal
Analysis (TGA) stability of the sample

Particle Size Analyzer To determine the particle size

(PSA) distribution

Tensile testing To determine the samples

response to the stress applied

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3.3 Flow of experiment

The figure below shows the experimental flow chart of sample preparation,
cellulose extraction and transparent thin film preparation process.

Sample preparation

OPEFB will be washed with distilled water, de-waxed using soxhlet extractor, autoclaved
with 10% NaOH, then suspended in 10% H₂O₂ and placed in orbital shaker

Cellulose extraction process

The fibre will be delignified with mixture of 20% formic acid and 10% hydrogen peroxide
(1:1) at 85°C for 60, 90, 120 min and bleached with 10% H₂O₂ for 90 min at 60°C

Preparation of transparent cellulose thin film solution

3g of extracted cellulose will be suspended in 100ml DMAc, added with 10g LiCl, heated
for 3 hours at 80°C and leaved overnight at 50°C

Preparation of transparent cellulose thin film

The solution will be casted on glass slide, left at ambient temperature for gelation, then
will be immersed in distilled water and dried in freeze-dryer

Sample analysis

The cellulose extracted and the film prepared will be analysed using TGA, FTIR, PSA
and tensile test

Figure 3.1: The flow of experiment in this research

20
3.4 Sample preparation

3.4.1 Washing process

100g of 500 microns OPEFB were soaked in 800 ml beaker and washed
several times with distilled water using Buchner funnel vacuum pump until the water
observed become clear. The purpose of washing is to remove the dirt and other
impurities from raw materials. Then, the fibres were dried in conventional at 40°C until
constant weight is obtained.

3.4.2 Pre-treatment of Oil Palm Empty Fruit Bunch (OPEFB)

12g of fibres were de-waxed with 120 ml of 70% Ethanol at ratio of fibre to
solvent 1:10 (g L-1) using Soxhlet extractor for 3 hours. The de-waxed fibres then were
collected and washed with distilled water for several times to remove the alcohol traces
before further drying in conventional oven at 40°C for overnight. The process were
repeated for several times until total of 50 g dried fibres is obtained.

50g of the de-waxed fibres were suspended in 10% of 100 ml NaOH and
autoclaved for one hours at 121°C and 1.5 atm. Dark supernatant is obtained. The
mixture of supernatant and fibres were separated and filtered using Buchner funnel
vacuum pump, followed by several round of washing until clear water was observed.
The treated fibres then were suspended in 100 ml of 10% H2O2 in 500 ml Erlenmeyer
flask and were placed into orbital shaker for 1 hours for bleaching treatment. The
bleached fibres then were collected and filtered before further washed with distilled
water and dried in conventional oven at 40°C for overnight.

21
3.5 OPEFB-Cellulose Extraction Process

The extraction method were adapted from Nazir et al. (2013). 15g of pre-
treated fibres were placed in three different 250 ml Erlenmeyer flask. The fibres were
soaked with the mixture of 200 ml of 20% formic acid and 10% H2O2 at ratio 1:1 (v/v).
Then, the soaked fibres were placed in water bath at 85°C at different reaction time
which is 60 minutes, 90 minutes and 120 minutes each for delignification process. The
delignified fibres then were collected, filtered and washed with 10% formic acid before
further rinse with distilled water until pH 7 were obtained. The extracted cellulose
observed is light yellow in colour. The extracted cellulose then were re-suspended in
50 ml of 10% H2O2 each for 90 minutes at 60°C for bleaching process. The process
were repeated for several times until the extracted cellulose turns white. Lastly, the
white suspension of extracted cellulose is obtained. The cellulose then were filtered
and washed several times before the insoluble fraction of cellulose were collected and
the yield (w/w) were calculated. The α-cellulose content were determined using TAPPI
method.

3.6 Transparent Cellulose Thin Film preparation

3.6.1 Preparation of Cellulose solution

3g of extracted cellulose at reaction time 60 minutes were suspended in

100ml of DMAc at concentrations of 5% w/v. The suspension were heated at 80°C
under stirring condition for 30 minutes. 8g of LiCl then were added to the cellulose
suspension after 30 minutes. The mixture continue heated at 80°C under stirring
condition for another 3 hours. The heat were decreased to 50°C at the end of heating
and the temperature were maintained for overnight. The same preparation process
were repeated for extracted cellulose at 90 and 120 minutes of reaction time.

22
3.6.2 Casting method to prepare transparent cellulose thin film

Certain amount of cellulose solution prepared were casted on casting glass

slide and were gently pressed with other glass slide to spread the solution uniformly
between the glass slides to prepare the film. The glass slides that were spread with
the solution were left for gelation at ambient temperature for overnight. After gelation
was formed, the glass slides contain the film were immersed in distilled water for
generation. The water were exchanged every 2 hours for 3 days to regenerate the film
and to remove the solvent. The wet film obtained were dried in freeze-dryer for 4 days.
The freeze-dried film then undergo further characterization.

3.7 Sample characterization

3.7.1 Fourier-Transform Infra-Red Spectroscopy (FTIR)

The functional group and chemical bonding of the surface structure are
usually determined by using FTIR. The infrared (IR) radiation will penetrated through
the sample. The sample will absorbed some of the IR radiation and generate
interferogram which will be represented in wave-like pattern for intensity versus time.
The fourier transformation then will transformed the transmitted interferogram into
spectrum. The FTIR in this research were done using Perkin Elmer Spectrum 400 FT-
IR/FT-NIR within wavelength of ranged 600-4000 cm-1 at a rate of 0.02 cm-1 to identify
the functional group of the cellulose extracted and the transparent thin film prepared.
Approximately 0.01g of sample were weighted and scattered in diamond ATR crystals.
A spectrum were recorded for each samples using air as a reference. The diamond
crystals will be cleaned using tissue and immersed in pure distilled water between the
consecutive sample runs.

23
3.7.2 Thermogravimetric Analysis (TGA)

TGA were used to carry out non-isothermal temperature programmed thermal

studies. The interaction between the gas and solid sample could lead to mass changes
(weight loss or weight gain) of solid samples at any controlled temperature, hence
thermal profiles is created. TGA were done using Mettler Toledo TGA machine (model:
TGA/SDTA 85-F) to determine the thermal stability of cellulose extracted as well as
the transparent thin film prepared. 15 mg of powder cellulose with particle range of 100
μm were weighted and placed in crucible and added to balance pan. Nitrogen gas were
flowed at 10 ml min-1 and heated from room temperature to 600°C with heating rate of
10°C min-1. Once the samples is fully decompose, the decomposition temperature of
each samples can be measured.

3.7.3 Particle Size Analyzer (PSA)

The Particle Size Analyzer is used to characterise the size distribution of

particles size in given sample. When the beam of light or a laser is scattered by a group
of particles, the angle of scattered light is inversely proportional to the particle size,
thus the desired particle size of the sample is obtained. The test were run using Particle
Size Analysis (Mastersizer, Malvern 3000). The temperature set were at room
temperature and the refrective index used is 1.746 with 1.000 absorption index. The
dispersant medium used is water with the dispersant refrective index 1.330.

3.7.4 Tensile testing

24
 CHAPTER 4

EXPECTED RESULTS

4.1 Expected Result

By the end of the study, the optimized parameters such as pH, acidity and
speed of the process will be finalized; hence the extraction is finalized. The cellulose
will be able to be extracted from OPEFB by using soda-pulping process and the
nanocellulose are able to be synthesised using acid hydrolysis from the extracted
cellulose. The transparent nanocellulose thin film will be prepared by casting method.
Then, several analysis will be conducted such as SEM, TEM, XRD, FTIR and TGA.

The colour of extracted cellulose are expected to change from brown to white
after alkali bleaching treatment. The surface morphology of the cellulose,
nanocellulose and transparent thin film will be identified in 3 dimension using SEM.
The extracted and treated cellulose including the nanocellulose and transparent thin
film are expected to have a smaller diameters and smoother surfaces compared to the
raw OPEFB. The changes for both colour and morphology to become lighter and
smoother is due to the removal of hemicellulose, lignin and other extractives from the
treatment and process.

The internal structure such as the crystallinity of the sample are identified in
2 dimension using TEM. The raw OPEFB are expected to shows unexposed crystalline
region surrounded with amorphous hemicellulose and lignin whereas the extracted

25
cellulose is expected to have the presence of large crystalline region with small
amorphous region. For the nanocellulose and transparent thin film, the result are
expected to have higher crystalline region intact and reduction in size of the fiber to
nanometer scale compared to the cellulose.

The crystallinity index of the samples will be able to determine using XRD.
The raw OPEFB are expected to have the lowest crystallinity index as it has high
amount of amorphous regions and the value are expected to increase after the
cellulose is extracted and treated using alkali bleaching treatment sue to the removal
of lignin and hemicellulose attached to the cellulosic fibers. The nanocellulose and
transparent thin film are expected to have the highest crystallinity index as acid
hydrolysis indicates the dissolution of an amorphous region of the cellulosic fibers.

The FTIR results are expected to shows changes in the functional group along
the process due to the various treatment. The lignocellulosic fibers consists of
hemicellulose, lignin and cellulose that are mainly composed by alkanes, esters,
ketones and alcohols. Table 4.1 below shows the typical functional group of cellulose
absorption peaks expected to appear after the process. At peaks 865 cm-1, it attributed
to the structure of cellulose due to ß-glycosidic linkages of the glucose ring and
cellulose chain.

Table 4.1 Typical functional group of cellulose absorption peaks

Functional group Peak, cm-1

CH2 bending 1432

CH bending 1375

C-O stretching 1326

C-H groups 865

The thermal stability of the structure are able to determine using TGA.
Lignocellulosic materials is a composite structure that will degrade below 400°C. Thus,
it is expected that the hemicellulose, lignin and cellulose will degrade at 180°C, 400°C

26
and around 300°C respectively. The thermal stability of the samples are expected to
increase along with the process due to the increased in crystallinity and intermolecular
hydrogen-bonded after the removal of amorphous content.

4.2 Research Costing

The cost of the materials required for this research is stated in Table 5.1. The
costing is calculated based on the price per unit mass or volume of each ingredients.

Table 4.2 The cost for each material per unit mass or volume (kg/L)

Amount Price (RM/kg

Materials Total price
needed or L)
Oil Palm Empty Fruit
5000 g 0.66 3.30
Bunch (OPEFB)
Sodium hydroxide
1200 ml 30.00 36.00
(NAOH)
Hydrogen peroxide
100 ml 50.00 5.00
(H2O2)
Sulphuric acid
105 ml 414.00 43.47
(H2SO4)
Dimethyacetamide
100 ml 686.00 68.6
(DMAc)

Lithium chloride (LiCl) 8g 771.00 6.17

TOTAL 162.54

27
4.3 Research Planning

The plan of this research is calculated to be done in the span of Semester 6

and Semester 7. The research will start on the first week in Semester 6 and expected
to be completed by the end of Semester 7 on the 17th week. The detailed of the
planning is according to the Gantt chart below.

28
 4.3.1 Gantt Chart

Table 4.3 Final Year Project Gantt Chart

FYP 1 FYP 2
No. Activities Jan- Feb- Mar- May- Jun- Jul - Aug- Sep- Oct- Nov- Dec-
Apr-20
20 20 20 20 20 20 20 20 20 20 20
1 Literature Survey

2 JSA preparation

3 JSA submission/approval

4 Proposal summary preparation

5 Proposal summary submission

6 Proposal summary presentation

7 Proposal preparation

8 Proposal submission

9 Cellulose extraction

10 Nanocellulose preparation

11 Transparent thin film preparation

12 Analytical tests

29
13 Thermal analysis

14 Morphology analysis

15 Experiment data collection

16 Result analysis

17 Writing thesis

18 Project presentation

19 Submission of Thesis (hard copy)

30
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