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Title of Project

Optimization of Solvent extraction of Phytonutrient from Palm Oil


Unripe Fruit by using Response Surface Methodology

Students Name: Damien s/o Susai


ID Number: 55201113147
Section: 6 BCP

Supervisors Name
EN. ZAINAL ABIDIN YUSOFF

Malaysian Institute of Chemical And


Bioengineering Technology
University Kuala Lumpur

07 October 2015

CHAPTER 1: INTRODUCTION

1.1 Introduction
Phytonutrients (also referred to as phytochemicals) are compounds found in
plants. They serve various functions in plants, helping to protect the plant's
vitality. For example, some phytonutrients protect the plant from UV radiation
while others protect it from insect attack.
Not only do phytonutrients award benefit to the plants but they also provide
benefits to those who enjoy plant food. That's because they have healthpromoting properties including antioxidant, anti-inflammatory, and liverhealth-promoting activities.
Fruits and vegetables are concentrated sources of phytonutrients; other plant
foods like whole grains, legumes/beans, nuts and seeds, and herbs and
spices also contain phytonutrients. Since many phytonutrients also serve as
the pigment that gives foods their deep hues, you can identify many
phytonutrient-rich foods by looking for colorful foods; for example, look for
foods that are blue or purple like blueberries, blackberries and red cabbage
(rich in flavonoids); yellow-orange foods like carrots, winter squash, papaya,
and melon (rich in beta-carotene); red or pink foods like tomatoes, guava,
and watermelon (rich in lycopene); and green foods like kale, spinach, and
collard greens (rich in chlorophyll). Yet, since not all phytonutrients give color,
it's important to not overlook some off-white foods as wellfor example,
garlic, onions, and leeks are rich in powerful sulfur-containing phytonutrients.
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1.2 Background Studies


The oil palm (Elaeis guineensis) is native to many West African
countries, where local populations have used its oil for culinary and other
purposes. Large-scale plantations, established principally in tropical regions
(Asia, Africa and Latin America), are mostly aimed at the production of oil,
which is extracted from the fleshy mesocarp of the palm fruit, and endosperm
or kernel oil. Palm oil is different from other plant and animal oils in that it
contains 50% saturated fatty acids, 40% unsaturated fatty acids, and 10%
polyunsaturated fatty acids. The fruit also contains components that can
endow the oil with nutritional and health beneficial properties. These
phytonutrients include carotenoids (alpha-,beta-,and gamma-carotenes),
vitamin E (tocopherols and tocotrienols), sterols (sitosterol, stigmasterol and
campesterol), phospholipids, glycolipids and squalene. In addition, it is
recently reported that certain water-soluble powerful antioxidants, phenolic
acids and flavonoids, can be recovered from palm oil mill effluent. Owing to
its high content of phytonutrients with antioxidant properties, the possibility
exists that palm fruit offers some health advantages by reducing lipid
oxidation, oxidative stress and free radical damage. Accordingly, use of palm
fruit or its phytonutrient-rich fractions, particularly water-soluble antioxidants,
may confer some protection against a number of disorders or diseases
including

cardiovascular

disease,

cancers,

cataracts

and

macular

degeneration, cognitive impairment and Alzheimer's disease. However, whilst


prevention of disease through use of these phytonutrients as in either food
ingredients or nutraceuticals may be a worthwhile objective, dose response
data are required to evaluate their pharmacologic and toxicologic effects. In
addition, one area of concern about use of antioxidant phytonutrients is how
much suppression of oxidation may be compatible with good health, as toxic
free radicals are required for defence mechanisms. These food-health
concepts would probably spur the large-scale oil palm (and monoculture)
plantations, which are already seen to be a major cause of deforestation and
replacement of diverse ecosystems in many countries. However, the
environmental advantages of palm phytonutrients are that they are prepared
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from the readily available raw material from palm oil milling processes. Palm
fruit, one of only a few fatty fruits, is likely to have an increasingly
substantiated place in human health, not only through the provision of
acceptable dietary fats, but also its characteristic protective phytonutrients.

1.3 Problem Statement


Malaysia is one of the biggest exporters of palm oil, contributes about 51% of
the worlds edible crude palm oil and accounts for 62% of world exports. In
the processing of palm oil, there are two types of products: crude palm oil
(CPO) and palm kernel oil (PKO). However, the major problem with this
process is the huge amount of wastes produced in the form of oil palm empty
fruit bunches (OPEFB), mesocarp fiber (MS), decanter cake (DC), and palm
oil mill effluent (POME) solids. Abundant oil palm unripe fruit bunches
(OPUFB) generated from the palm oil mill industry create huge problems for
the environment and the palm oil mill itself. In this project, two methods will
be used which is microwave assisted extraction and ultrasonic extraction.
Thus, comparative study will be made on which extraction will give out a
better quality as well as high yield of the essential oil extracted.

1.4 Research Objectives


The objectives of this project are:
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To extract Phytonutrients from Palm Oil Unripe Fruit using


solvent extraction method
To study the efficiency on the extraction based on solvent ratio,
time of extraction, temperature and type of solvents
To analyse Phytonutrients by using UV-VIS Spectroscopy
To study the optimum parameters for solvent extraction of
Phytonutrients by RSM method

CHAPTER 2: LITERATURE REVIEW

2.1 Phytonutrient
There has been a growing research interest in palm, especially palm
oil, which is one of the major edible plant oils in the tropical countries. The oil
palm (Elaeis guineensis) is native to many West African countries, where
local populations have used its oil for culinary and other purposes. Largescale plantations, established principally in tropical regions (Asia, Africa and
Latin America),are mostly aimed at the production of oil, which is extracted
from the fleshy mesocarp of the palm fruit, and endosperm or kernel oil. Palm
oil is different from other plant and animal oils in that it contains 50%
saturated fatty acids, 40% unsaturated fatty acids, and 10% polyunsaturated
fatty acids. The fruit also contains components that can endow the oil with
nutritional and health beneficial properties. These phytonutrients include
carotenoids
tocotrienols),

(-,

and

sterols

-carotenes),

(sitosterol,

vitamin

stigmasterol

(tocopherols

and

and

campesterol),

phospholipids, glycolipids and squalene. In addition, it is recently reported


that certain water-soluble powerful antioxidants, phenolic acids and
flavonoids,

can

be

recovered

from

palm

oil

mill

effluent.

2.2 Palm Oil Unripe Fruit


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Oil palm fruit is typically grown in the tropical areas in South East Asia,
South Africa and South America . Palm oil provides vital food for millions of
people and has been found to be very healthy for the human diet. Palm oil
has good resistance to oxidation and prolonged exposure to high
temperatures, thus making it ideal for frying. A high percentage of palm oil is
usually added to frying oils because of its structure and other economic
advantages. The Malaysian palm oil industry is considered to be highly
regulated. A major problem faced by oil palm exporters and producers is the
accurate grading of fresh oil palm fruits according to their ripeness levels
before processing. The maturity or ripeness of the oil palm fruits dictates the
quality as well as overall marketability of the palm oil produced .Oil palm
fruits can generally be categorized into four ripeness grades: ripe, underripe,
unripe, and overripe . Bunches range in color from yellow, reddish orange,
red, to black. A fruit before maturity is typically yellow at the base and dark
purple to black at the apex. A young palm has 50 to 100 red-violet ripe fruits
per bunch. The ratio of oil palm fruit pigments, such as carotenoids and
chlorophylls, affects the color of the oil palm fruit. For example, unripe fruits
have a higher proportion of chlorophyll that gradually decreases upon
maturity . Similarly, carotenoids increase as oil palm fruits mature . Color
changes resulting from biochemical reactions can likewise be related to fruit
maturity.
2.3 Solvent Extraction
Various solvents have been used to extract different phytoconstituents. The
plant parts are dried immediately either in an artificial environment at low
temperature (50-60C) or dried preferably in shade so as to bring down the
initial large moisture content to enable its prolonged storage life and . The
dried berries are pulverised by mechanical grinders and the oil is removed by
solvent extraction. The defatted material is then extracted in a soxhlet
apparatus or by soaking in water or alcohol (95% v/v). The resulting alcoholic
extract is filtered, concentrated in vacuo or by evaporation, treated with HCl
(12N) and refluxed for at least six hours. This can then be concentrated and
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used to determine the presence of phytoconstituents. Generally, the saponins


do have high molecular weight and hence their isolation in the purest form
poses some practical difficulties. The plant parts (tubers, roots, stems, leave
etc) are washed sliced and extracted with hot water or ethanol (95% v/v) for
several hours. The resulting extract is filtered, concentrated in vacuo and the
desired constituent is precipitated with ether. Exhaustive extraction (EE) is
usually carried out with different solvents of increasing polarity in order to
extract as much as possible the most active components with highest
biological activity.
2.4 Response Surface Methodology (RSM)
Response surface methodology (RSM) is a collection of mathematical
and statistical techniques for empirical model building. By careful design of
experiments, the objective is to optimize a response (output variable) which
is influenced by several independent variables (input variables). An
experiment is a series of tests, called runs, in which changes are made in the
input variables in order to identify the reasons for changes in the output
response.

CHAPTER 3: METHODOLOGY

3.1 Chemicals, Materials and Apparatus


Chemicals: ethanol
Raw Materials: Palm Oil Unripe Fruit
Instruments: microwave digester, laboratory ultrasonic cleaner, UV-Vis
Spectroscopy, digital weighing scale, rotary evaporator

3.2 Summarized Methodology

Overall Procedures
Before

run

the

experimental

procedure,

Response

Surface

Methodology(RSM) was designed for the whole process. The experiment


was followed the schedule that designated by RSM. When the experiment of
solvent extraction was finished, the extracted oil of Phytonutrients was
analysed by using HPLC. The experiment was carried out by three important
parameters of control which are different time of solvent extraction,
temperature and ratio of plant material mass to volume of solvent.
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Preparation of sample solution


For extraction of phytonutrient by solvent process, unripe palm fruits were
extracted using the solvent (ethanol). The amount of unripe palm fruits used
is kept constant.

Microwave assisted extraction


Prepared sample will be introduced into microwave digester for about 30
minutes at the temperature of 55 degree Celsius.
Ultrasonic assisted extraction
At this stage, a comparative study have been made on the extraction kinetics
of phytonutrients with and without the ultrasonic waves. The setting of
frequency is to be determined.
Evaporation via rotary evaporator
Solution will be heated and evaporated to obtain solvent free oil at
temperature of 80 degree Celsius.
UV-VIS Spectroscopy
Compound solutions were prepared and tested with UV-VIS Spectroscopy.
Performed

analysis

to

calculate/identify

the

concentration

bioactive

component in the oil solution.

CHAPTER 4: RESULT

4.1 EXPECTED RESULT


One of the main objectives of this research is to study the efficiency on the
extraction based on solvent ratio and time of extraction and to extract
PHYTONUTRIENTS from Palm Oil Unripe Fruit using solvent extraction
method and to study the optimum parameters for solvent extraction of
Phytonutrients by RSM method investigated that ethanol produced higher oil
yield at optimum temperature of 55 degree Celsius. And also, ethanol gave
the maximum oil yield.

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CHAPTER 5: RESEARCH PLAN


2015
Au Sep
g
t

Oc
t

No

De
c

Ja
n

Fe
b

Ma
r

Ap
r

Ma
y

Proposal
Search related journal,
identify the method for the
project
Submission the first draft
of the proposal
Submission
of
the
proposal
Presentation
of
the
proposal
FYP Report
Writing the first chapter of
the report
Experiment
Prepare all the raw
materials, and equipment
Run the experiment
FYP Report
Writing the second and
third chapter of the report
Submission
of
draft
chapter 1,2 and 3
Finish writing the report
Submission of the first
draft
Presentation
Presentation of slides
FYP Report
Submission of the final
draft and hard cover

11

Ju
n

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