TABLE OF CONTENTS

LIST OF TABLES.............................................................................................................................iii
LIST OF FIGURE.............................................................................................................................iii
CHAPTER 1: INTRODUCTION.........................................................................................................1
1.1 Background of Study...........................................................................................................1
1.2 Problem Statement.............................................................................................................4
1.3 Objectives............................................................................................................................6
1.4 Scope of Study.....................................................................................................................6
1.5 Significant of Study..............................................................................................................7
CHAPTER 2: LITERATURE REVIEW..................................................................................................9
2.1 Lemongrass.........................................................................................................................9
2.1.1 Overview of Lemongrass..............................................................................................9
2.1.2 Advantages of Lemongrass.........................................................................................10
2.1.3 Properties of Lemongrass Oil.........................................................................................13
2.2 Extraction Method of Lemongrass Essential Oil................................................................16
2.2.1.1 Solvent Extraction...................................................................................................16
2.2.1.2 Steam Distillation....................................................................................................17
2.2.1.3 Hydro Distillation.....................................................................................................17
2.2.1.4 Ultrasonic Assisted Extraction.................................................................................18
2.2.2 Previous Study of Lemongrass Oil..............................................................................18
2.3 Factors Affecting Extraction..............................................................................................23
2.3.1 Size.............................................................................................................................23
2.3.2 Time...........................................................................................................................23
2.3.3 Ratio...........................................................................................................................24
2.4 Analytical Test...................................................................................................................24
2.4.1 Fourier Transform Infrared Spectroscopy (FTIR)........................................................24
CHAPTER 3: METHODOLOGY.......................................................................................................25
3.1 INTRODUCTION.................................................................................................................25
3.2 Raw Material.....................................................................................................................25

i
 3.3 Chemicals and Equipment Used........................................................................................25
3.4 Outline Procedures............................................................................................................26
3.5 Experimental Work............................................................................................................27
3.5.1 Sample Preparation....................................................................................................27
3.5.2 Extraction Process......................................................................................................28
3.5.3 Analysis of Sample......................................................................................................30
3.5.3.1 Fourier Transformed Infrared Spectroscopy...........................................................30
CHAPTER 4: RESULT AND DISCUSSION........................................................................................31
4.1 Introduction......................................................................................................................31
4.2 Experimental Design..........................................................................................................31
4.3 Ultrasonic Assisted Extraction...........................................................................................32
4.3.1 Time of Extraction......................................................................................................33
4.3.2 Sample to Solvent Ratio.............................................................................................35
4.4 Fourier Transformed Infrared Spectroscopy.....................................................................38

ii
 LIST OF TABLES
Table 2. 1 Components of lemongrass.................................................................................13

Table 2. 2 Previous Results of Different Method.................................................................19

LIST OF FIGURE
Figure 2. 1 Lemongrass............................................................................................................9
Figure 2. 2 Lemongrass Tisane.............................................................................................11
Figure 2. 3 Citral  and Citral .............................................................................................14
Figure 2. 4 Percentage Oil Yield for Solvent and Steam Distillation Extraction
(M.A.Suryawanshi, 2016)........................................................................................................20
Figure 2. 5 Effect of Time on Lemongrass Oil Hydro Distillation Extraction (Thien Hien
Tran, 2019)................................................................................................................................21
Figure 2. 6 Results for Ultrasonic Assisted Extraction with Different Parameters
(Mohamed Amine Balti, 2017)................................................................................................22
Figure 2. 7 Different Results for Different Size of Material (E.T.Akhihiero, 2016)..........23

iii
 CHAPTER 1: INTRODUCTION

1.1 Background of Study

Nowadays, citrus fruits and plants are not an insignificant name in this
world. Numerous research has been made on every and each type of citrus
plants. One of it is lemongrass or its scientific name Cymbopogon. Lemongrass
belongs to the grass family, specifically the Poaceae family of grasses [CITATION
Lis12 \l 1033 ]. There are other citrus plants like lemongrass such as lemons,
limes, and oranges. Citrus plants usually have strong essence and have many
applications. Lemongrass has more than 50 types, but the most common types
are the Cymbopogon citrus and Cymbopogon flexuosus. The one used in this
experiment are the Cymbopogon citrus as this type is the mostly cultivated in
Malaysia. On the other hand, Cymbopogon flexuosus is common in India and
Thailand. Lemongrass is colored yellowish-green and has an aromatic citrus
scent. It has a bulbous bottom and tastes like lemon as its name goes [CITATION
Wha19 \l 1033 ].

Lemongrass has a lot of application and each of it varies on how it is

used. It is used in pharmaceutical, therapy, and also in food production.
Lemongrass is rich with aromatic scent and rich in vitamin B and folate which
promotes the production of energy and red blood cells [ CITATION Bet12 \l 1033 ]. In
the past, lemongrass tea is a very exclusive serving for the executives in China
for its refreshing and healthy effects on the body and mind. Lemongrass herb
tea has a relaxing effect that can relief stress and improve the circulation of

1
blood in the body. Other than that, lemongrass is an important ingredient for a
cuisine that is well known throughout the world especially at the Asia continent.
The cuisine is Tom Yum, a sour-spicy coconut broth. A veery popular and loved
by everyone especially Asian people. It is well suited to be ate with rice and
noodles.

2
 In lemongrass essential oil, there is an average of 70-80% of a
component known as citral. Citral contained in the lemongrass has many of its
uses such as in perfumery section and also can be used in making synthetic
vitamin A [ CITATION Wil17 \l 1033 ]. Lemongrass composition is Citral-α (40.8%),
Citral-β (32%), Nerol (4.18%), Geraniol (3.04%), Citronellal (2.10%),
Terpinolene (1.23%), Geranyl Acetate (0.83%), Myrecene (0.72%) and Terpinol
(0.45%). There are also other components such as Pinene, Limonene and
Linalool but it all just only traces of it [ CITATION Gag11 \l 1033 ].

There is various of benefits that can be obtained by the uses of Citral in

nowadays economy. Citral is known as a main component in fragrance and
flavoring. Citral emits a refreshing and natural taste and scent. It makes a very
good perfume and gives flavors to any cuisine. Other than that, it has an
unbelievable curing effect on the body. It is very popular as antispasmodic,
analgesic and anti-inflammatory effect. Other than gives off therapeutic scent, it
also helps in relaxing muscles and relieving cramps. Furthermore, in certain
places, lemongrass extract is used to healing wounds and helps in recovering
bone fractures.

The method on extracting lemongrass essential oil is variety. There is the

method of hydrodistillation, microwave-assisted hydrodistillation, steam
distillation, and ultrasonic assisted extraction. Among all of this, the most
optimum method is by the ultrasonic assisted extraction (UAE) as it uses way
less energy during the extraction and helps in reducing the costs of extraction. It
also protects the components from thermal damage that can occur to the extract
during the process [ CITATION TWC16 \l 1033 ]. Nevertheless, the method used in
the industry in obtaining lemongrass essential oil is by steam distillation. This is
because there is no large size of ultrasonic extraction bath. Therefore, steam
distillation method is used in industrial process of extracting lemongrass
essential oil.

3
1.2 Problem Statement

From thousand of years before, human has been using various of method
in maintaining the stability of the economy. Lots of new products has been
invented and created and each of them has its own specialties. In this case,
lemongrass plays a very important role. The component contained in
lemongrass that is Citral has shown various application and helps in creating
new products [ CITATION Ale19 \l 1033 ]. Therefore, the demand of lemongrass
extract has been booming and highly searched by many companies.

The demands on lemongrass essential oil that is increasing in nowadays

economy has open a brand-new opportunity to manufacturing companies.
Company that process and extracts essential oil from lemongrass is gaining
more and more profits. In this case, various methods have been run to confirm
the best way on extracting the essential oil from lemongrass. This action has
made the analysis and experiment on which method that gives the most
advantages in it from the term of quality, cost, and product yield. Therefore, it is
important on identifying the method which giving the widest opportunity for a
company to success and have bigger profits.

The industrial method used in extracting essential oil from lemongrass is

by using steam distillation method. This method is easy to extract the essential
oils, but it has a few downsides on this method. The drawbacks that may have
occur when running this process is that there will be thermal damage on the
components of the extract [ CITATION RAN14 \l 1033 ]. As direct steam is flowed to
the raw material, the high temperature of steam may change the constituents of
the product. This change may look small, but it deters the components
percentage in the extract.

4
 The method that will be implemented in this experiment is the ultrasonic
assisted extraction (UAE). UAE is an easier and less cost method in extracting
the essential oil from the lemongrass. Other than that, it also does not affect the
components of the product. This making it more flexible and preferrable
[ CITATION Yol13 \l 1033 ].

5
1.3 Objectives

The objectives of this experiment were:

1. To extract the essential oil (citral) from lemongrass by using ultrasonic

assisted extraction method.
2. To evaluate the effects of material size, time of extraction, and sample to
solvent ratio towards the essential oil extraction.
3. To determine the functional group of the raw material used by using
Fourier-Transformed-Infrared Spectroscopy (FTIR) method.
4. To analyze the functional group of essential oil obtained by using Fourier-
Transformed-Infrared Spectroscopy (FTIR) method.

1.4 Scope of Study

To achieve the objectives if this research study in the investigation of

extracting lemongrass essential oil by using UAE method, this project was run at
Lab 1C, Pilot Plant, and Analytical Lab 2B at UniKL MICET, Alor Gajah, Melaka.
The UAE method is run with several parameters and oil yield was recorded from
each run. The effects of parameter such as size of raw material (grind and
chunk) , time of extraction (2,4, and 6 hours), and sample to solvent ratio (1:1,
1:2, 1:3) to the yield of oil extracted. The temperature used in this experiment is
at 60 °C and the solvent used is ethanol. The sample and extracted product
were also analyzed to confirm and investigate its components and constituents
inside each compound.

6
1.5 Significant of Study

The application of lemongrass essential oil gives a lot of to the daily life of
a person. The components contained in the oil helps in many sectors and
making life easier whether in pharmaceutical section, culinary or even daily
necessities. By extracting and processing lemongrass essential oil, a handful of
things can be done without needed to use the lemongrass stalk itself. More
advantages and other benefits can be applied through this method. Generally
known, lemongrass has an aromatic and refreshing scent. This scent is known
to help in relieving stress and assists in relaxation process [ CITATION Sul17 \l
1033 ]. Other than that, citronella is a component consists in the lemongrass oil.
Usually, lemongrass is extracted to receive this component. It has many uses
such as citrusy scent that repels mosquitoes and bugs [ CITATION Bob16 \l 1033 ] .
Moreover, it has a sweet citrus scent making it more comfortable for human to
use it.

Other than help in repelling bugs and mosquitoes, lemongrass is famous

in its uses in cooking especially Asian cuisines. The sweet and refreshing taste
of lemongrass has made innovation to many dishes in this world. Lemongrass’s
scent also higher the appetite for the people in eating the dishes made with it. In
Brazil, the stalk is dried and then is made into tea. Lemongrass tea is good for
digestion and antispasmodic [ CITATION Sim19 \l 1033 ] . Furthermore, lemongrass
has a component which has an anti-inflammatory effect. In certain country,
lemongrass is boiled with water and is taken orally. Lemongrass boiled water
acts as emmenagogue.

The method used in this experiment is ultrasonic assisted extraction. UAE

is preferred as it is more efficient and cost saving. Other methods such as hydro
distillation and steam distillation can be used too but it consumes too much
energy compared to UAE. Other than that, UAE also keeps the compounds to

7
be in good condition. On the other hand, steam distillation and hydro distillation
may cause thermal damage to the components of the essential oil extracted.
Other than that, UAE also gives of more yield than using steam and hydro
distillation method [ CITATION Kat20 \l 1033 ].

8
 CHAPTER 2: LITERATURE REVIEW

2.1 Lemongrass

2.1.1 Overview of Lemongrass

Cymbopogon citratus or generally known as lemongrass is an

herbaceous perennial plant. Came from the family of Gramineae, lemongrass is
popular and well known throughout the world. Common names such as barbed
wire grass, citronella grass and serai are one of the various calls of lemongrass
in other countries. Lemongrasses are mostly cultivated at Asia, Africa, and
Australia. There is also several planted in tropical island which has humid and
hot weather which is the optimum habitat for lemongrass . [ CITATION Mar121 \l 1033
]

Figure 2. 1 Lemongrass

9
 When talking about lemongrass, a light green stem with citrus scent will
be imagined. The stems are multiple and tender as it is an herbaceous perennial
plant. Herbaceous is a term for a plant which have no woody stems while
perennial means a plant that have a life term more than two years.
Lemongrasses cultivate in a humid and warm region. As for the survival of
lemongrass, after its growing season the plants will die at the end of the growing
season. In this case, new lemongrass will grow from the stems that left under
the ground. In normal occasion, four seasons country can cultivate lemongrass
only on summer and spring season only while during winter and autumn, the
stem while stay under the ground waiting for the next season to come and grow
a new piece [ CITATION Lem \l 1033 ].

2.1.2 Advantages of Lemongrass

Lemongrass has a long history in pharmaceutical section. Various

medicine can be made from lemongrass whether with addition of other
substances or using it right off the bat. For several distinct situations,
lemongrass is usually taken orally, rubbed directly to the skin, or inhaled as
aromatherapy. But to support some of its traditional applications, there is little
science evidence. For instance, the extraction of lemongrass oil has a more
concentrated content of lemongrass’ nutrients than using it directly. Lemongrass
oil benefits in skincare treatment as it has personification of detoxifying and
purifying. Various brands have utilized these properties of lemongrass such as
Garnier and Eminence [ CITATION Lem19 \l 1033 ] . These brands are top local
brands in our country and have many of its buyers from all over the world. This
shows that using lemongrass as base for skincare has many of its benefits.

10
 Figure 2. 2 Lemongrass Tisane
Other than skincare and medication, lemongrass also has been used as
flavoring especially, in food and beverages. Lemongrass has citrusy aroma
which similar to any citrus fruits such as lemons and oranges. Its flavor is unique
as it is like lemon but with the trails of ginger at the aftertaste. This eccentric
flavor and aroma are the perfect combination for making any food. Most popular
foods that utilized the uses of lemongrass is Tom Yum, Padprik and any seafood
dishes. The intriguing flavor of lemongrass packs a punch in the savoring dish.
For beverages, lemongrass is known in making tisanes or generally known as
herbal teas. In China, lemongrass tea is believed to balance the yin and yang
energy inside one body [ CITATION Ger01 \l 1033 ] . Claiming a patient with unbalanced
yin and yang energy will have various illness until the energy is balanced. On
the scientific discussion, lemongrass tea proved to heal the body by
detoxification. As lemongrass is natural diuretic, the metabolism of body
increased and helps in detoxify the body. Natural diuretic properties are the
increase of urine produced in body which enables in detoxification.

11
 In industries, lemongrass often used as fragrance scents in soaps and
deodorants. The refreshing aroma of citral inside lemongrass gives the wearers
best feeling when using it. Other than that, lemongrass oil is believed to remove
dandruff in hair scalps. Its antimicrobial and inflammatory properties decrease
the symptoms of dandruff. In findings, lemongrass tonics that contains 10% of
lemongrass oil helps in reducing dandruff by 81% [ CITATION Jam19 \l 1033 ].
Lemongrass has the component terpinene-4-ol that possesses antimicrobial
properties and block the growth of bacteria in scalps. Therefore, it is used widely
in industries and variants of toiletries manufactured has these properties.

12
2.1.3 Properties of Lemongrass Oil
Lemongrass is called citrus plant due to its high components of Citral.
Other than that, there is also other components that contained inside
lemongrass. Table 2.1 shows the list composition/compound in lemongrass
(citation)

Components of lemongrass Percentage of components (%)

Citral  40.8
Citral  32
Nerol 4.18
Geraniol 3.04
Citronellal 2.10
Terpinolene 1.23
Geranyl acetate 0.83
Myrecene 0.72
Terpinol 0.45
Methylheptenone 0.2
Borneol 0.1–0.4
Linalyl acetate 0.1
 Pinene 0.07
 Pinene 0.04
Limonene Traces
Linalool Traces
-caryophyllene Traces
Table 2. 1 Components of lemongrass

13
 Lemongrass consists of more than half citral inside it. There are two types
of citral inside lemongrass that is citral  and citral . These two compounds are
actually geometric isomers. Citral  is an E-isomer that is called geranial while
citral  is a Z-isomer known as neral

Figure 2. 3 Citral  and Citral  (citation)

Citral is the source for the strong lemon odor inside of lemongrass. Its
appearance is a pale-yellow liquid. This component is used in perfumery as the
odor at soaps, deodorant, and perfumes. The sweet and relaxing sensation of its
scent is interesting and suit for any perfumery products. Other than that, citral
also used in food and beverages as flavor. Citral fortifies and add the lemon-like
flavor to servings. Moreover, it has strong microbial qualities and used for
treatment in several country. Other usage of citral is in the manufacturing of
vitamin A. It is used to mask the smell of antibiotics and medicinal substances
[ CITATION EMe89 \l 1033 ].

14
 Referring to previous study, there are various studies on the
pharmacology activities on lemongrass. There are a lot of good effects of
lemongrass compositions and a lot more to be discovered. Lemongrass has
anti-amebic effect, antibacterial and antidiarrheal effects. The composition that
responsible on these effects are geranial and neral or known as citral  and
citral . Lemongrass is packed with various potential therapeutic effects and
profitable for the community [ CITATION Gag16 \l 1033 ].

In food manufacturing, dried lemongrass is crushed and grinded into

powder. This powder is seasoned and utilized as a seasoning for food
processing. The fragrant and bittersweetness flavor of the powder is fit for any
dish. Other than giving eccentric flavor, lemongrass powder also contains
therapeutic properties and can be categorized as healthy diet. This powder
traditionally used in several country such as Malaysia, Thai, and India [ CITATION
Syl13 \l 1033 ].

Lemongrass extract also can be utilized as insect repellent. Geranial

(citral ) and neral (citral ) is effective in repelling insects especially mosquitoes
and house fly. There is also geraniol and citronellol that known as repellent
compounds, but it presents in a small amount in the lemongrass extract. There
is past investigation on the activity of mosquitoes on lemongrass extract-soaked
blood pad and the otherwise. The investigation shows that the activity of
mosquitoes on blank blood-soaked pad is higher than on lemongrass extract-
soaked blood pad. The exposure of lemongrass extract to the pad disturbs the
mosquitoes and their activity is more concentrated on the blank blood soaked
pad [ CITATION Bal13 \l 1033 ].

15
2.2 Extraction Method of Lemongrass Essential Oil
The process of extraction is crucial to gain the essential oil of a part of
plant. Lemongrass essential oil is separated from the lemongrass plant through
the process of extraction. There are many extraction approaches to separate
essential oil from lemongrass. To extract essential oil from lemongrass, various
method can be used such as solvent extraction, steam distillation, hydro
distillation, and ultrasonic assisted extraction [ CITATION MAS16 \l 1033 ]. Each
method gives different results and yield of essential oil ad more ways are still in
research for the optimum results of extraction.

2.2.1.1 Solvent Extraction

Solvent extraction is an extracting method where lemongrass will be

sliced into smaller parts to increase its area of surface and placed into a flask
containing of hexane solvent. The content of the flask is then will be left for 36
hours that is a day and half. The extract in the lemongrass will be extracted
completely by then. After 36 hours has passed, the lemongrass will be
separated from the solvent and ethanol solution will be added to the solvent.
Essential oil is soluble in ethanol solution as ethanol is alcoholic solution. Next,
the solution will be transferred to a separating funnel and undergo a separating
process named liquid-liquid separation. After a few hours, two layers of solution
will be visible as the density of each solution is different. The bottom layer is the
ethanol and lemongrass extract while the top layer is the hexane solution.
Lastly, the ethanol solution is separated from the extract in the rotary
evaporator. The water bath temperature will be set at 78°C due to the
evaporation temperature of ethanol is that [ CITATION MAS16 \l 1033 ].

16
2.2.1.2 Steam Distillation

Distillation is a separating process in which a mixture is vaporized and

then condensed to acquire the liquid demanded. Steam distillation is a process
where hot steam is flowed through the sample to extract the components
inquired. The vapor will then flow into a condenser and will be condensed. The
condensed solution will be collected and transferred into a separating funnel.
Two forms of layer will be visible due to the difference in density of the two
solution. At the bottom layer is water while on the top layer is the essential oil
extracted. There will be a tap at the separating funnel and water will be
extinguished from the funnel. Once the water is fully discarded, essential oil can
be collected in another bottle or volumetric flask. The container of the oil needs
to be fully sealed to avoid the vaporization of essential oil [ CITATION MAS16 \l
1033 ].

2.2.1.3 Hydro Distillation

Hydro distillation is a method which has the same principle as steam

distillation but with a different method. As for steam distillation, steam is flowed
through the sample but for hydro distillation, the sample is boiled with the
solvent and the vaporized solvent is condensed. Method wise speaking, the
lemongrass will be boiled in deionized water and vapor will be condensed and
collected. Hydro distillation is usually done with a Clevenger apparatus. The
vapor flow into the Clevenger apparatus will be condensed with the stream of
cooling water. This process usually takes about 6 hours depending on the
sample. 100g of lemongrass sample took about 6 hours for the optimum result
[ CITATION Thi191 \l 1033 ].

17
2.2.1.4 Ultrasonic Assisted Extraction
Ultrasonic assisted extraction is a method where ultrasonic is used to
extract essential oil relating its high frequency wave in the bath. These waves
vibrate through the sample penetrating the compound and extracting it. The
sample will be simmered in ethanol solution as lemongrass compounds are
soluble in alcoholic solution. Implying heat to the process also helps in
shortening the extraction time. The compound contained inside solid sample will
be extracted into liquid phase [ CITATION Moh17 \l 1033 ] . This method is
inexpensive, simple and has green technology application. Other than that, this
method lowers the probability of thermal degradation by far compared to the
other methods.

2.2.2 Previous Study of Lemongrass Oil

There are various types of method can be used to process and extract
lemongrass essential oil. Each method has its own advantages and
disadvantages. By comparing each and every investigation of method used to
extract, the best method that gives the highest yield can be determined.
Therefore, previous study of lemongrass essential oil extraction method will be
compiled and compared to decide the best method of extraction.

18
Method Raw Material Parameter Result Reference
Solvent - Dried lemongrass Time: 36 hours of absorption time. Mass oil yield: 1.9g out of 150g [ CITATION MAS16 \l
Extraction - Hexane solution 20 hours of extraction time. sample. 1033 ]
- Ethanol Temperature: 78°C during % oil yield: 1.30% out of 150g sample.
separation process.
Steam - Dried lemongrass Time: 20 hours of extraction time. Mass oil yield: 1.01g out of 150g [ CITATION MAS16 \l
Distillation - Distilled water Temperature: 100°C sample. 1033 ]
% oil yield: 0.71% out of 150g sample.

Hydro - Dried lemongrass Time: 30, 60, 90, 120, 150 minutes Mass oil yield: 0.35g out of 100g [ CITATION Thi191 \l
Distillation - Deionized water of extraction time. sample. 1033 ]
Temperature: 100°C % oil yield: 0.35% out of 100g sample.

Ultrasonic - Dried lemongrass Time: 5 hours of extraction time. Mass oil yield: 3.093g out of 100g [ CITATION Moh17 \l
Assisted - Ethanol Temperature:30, 40, 50, 60°C sample. 1033 ]
Extraction Power: 70, 160, 250W (at 40kHz) % oil yield: 3.093% out of 100g sample.

Table 2. 2 Previous Results of Different Method

19
Table 2.2 show the previous study of lemongrass oil extraction using different
methods. M. A Suryawanshi 2016 reported that by using solvent extraction
method resulted about 1.3 % yield of lemongrass oil solvent extraction method
yields more percentage of essential oil compared to steam distillation method.
Using the same mass of sample that is 150g of dried lemongrass, solvent
extraction yields 0.6% more oil yield against steam distillation that is 0.70%.
Other than having more yield oil than steam distillation, solvent extraction is more
superior than steam distillation as the essential oil extracted will be less exposed
to thermal degradation, hydrolysis, and water solubilization of some compounds
[ CITATION MAS16 \l 1033 ].

20
 Figure 2. 4 Effect of Time on Lemongrass Oil Hydro Distillation Extraction
[ CITATION Thi191 \l 1033 ].

The extraction of lemongrass essential oil has been run and recorded in
the Indonesia Journal of Chemistry written by Thien Hien Tran (year). The figure
2.5 shows the yield of lemongrass essential oil collected with different time count
of extraction. The sample mass used is 100 g of dried lemongrass and the
highest yield recorded is 0.35%. Result shows that the oil does not yield more
than 0.35% even with different time of extraction. Theoretically, the longer
extraction time, the higher oil yield returned. However, the amount collected when
the time is extended still does not increase and the oil quality collected may be
affected if exposed to temperature for a long period [ CITATION Thi191 \l 1033 ].
According to a website article, “How Temperature Affect Essential Oils”, if an
essential oil is exposed to heat, it will automatically lose its medicinal qualities
[ CITATION Tas18 \l 1033 ].

21
 Figure 2. 5 Results for Ultrasonic Assisted Extraction with Different Parameters
[ CITATION Moh17 \l 1033 ].

The results for ultrasonic assisted extraction differ by its parameter. Based
on the result above collected from The Chemical Engineering and Processing
Journal written by Mohamed Amine Balti, the most compound extracted during
the investigation is at E4 with a maximum of 23 compounds. The parameter for
E4 is the highest ultrasonic power that is 250W, 5 hours of extraction and with
temperature of 30°C. The highest mass oil yield is 3.093 g out of 100 g sample
that is 3.093%. This result shows that ultrasonic assisted extraction deems the
best method on extracting essential oil [ CITATION Moh17 \l 1033 ]. Ultrasonic waves
break down the cell wall of lemongrass and improve solvent penetration and
results in better extraction of targeted sample [ CITATION BKT12 \l 1033 ].

22
2.3 Factors Affecting Extraction

2.3.1 Size

It is believed that smaller size of raw material yields more essential oil
during extraction [ CITATION Kir13 \l 1033 ]. An investigation has been made about
how size of raw material affects the yield of extraction.

Figure 2. 6 Different Results for Different Size of Material [ CITATION ETA16 \l 1033 ].

Having small particles of material increases the yield of essential oil

extracted. This is due to there is more area exposed by being small particle
rather than bigger particles. Other than that, the time of extraction will be
shortened too due to small particles material oil cells will be extracted far quickly
than bigger particles [ CITATION Sar17 \l 1033 ].

2.3.2 Time

Time is a big factor that affects the extraction process. Longer time of
extraction yields more essential oil than short duration of extraction. However,
when dealing with heat, duration of extraction needs to surveil. This is because
long expose to high temperature may have an effect to the composition of the
essential oil [ CITATION Val14 \l 1033 ] . A research has been made about effect of
time to oil yield. In this case, the material undergoes extraction process with two
different duration that is 20 minutes and 80 minutes. From the result, 80 minutes
extraction has more oil yield that is 0.338% compared to 20 minutes time,
0.282% [ CITATION Val14 \l 1033 ].

23
2.3.3 Ratio

Sample to solvent ratio plays a big role in optimizing the oil yield during
extraction. By increasing the solvent ratio, total yield of essential oil can be
increased as well. This phenomenon caused by the concentration gradient
between the sample and solvent became greater that leads to good mass
transfer among the composition [ CITATION Sep09 \l 1033 ].

Please explain further!!

2.4 Analytical Test

2.4.1 Fourier Transform Infrared Spectroscopy (FTIR)

Fourier Transform Infrared Spectroscopy or in short FTIR is a method on

analyzing the chemical properties of a material whether organic or inorganic. By
sending a range of infrared radiation through a sample, it can be detected the
radiation which is absorbed by the sample and radiation that managed to pass
through. By this, the molecular identification of compounds can be obtained. The
range of infrared radiation send is about 4000 to 400cm -1 [ CITATION FTI15 \l 1033 ].

Other than analyzing chemical compounds, FTIR seldom used to identify

contamination, additives, and oxidation in a material. It also can detect
decomposition or uncured monomers in a material. It is a simple and effective
way on analyzing materials and chemical composition. All material is suitable for
FTIR analysis except for homo-nuclear diatomic molecules due to its infrared
inactive status.

24
 CHAPTER 3: METHODOLOGY
3.1 INTRODUCTION
In this chapter, the procedures and method of extraction essential oil from
lemongrass carried out during the experiment will be described and elaborated.
The method used in this extraction process is the Ultrasonic Assisted Extraction.
The experiment started by preparing the raw material needed that is lemongrass
and the composition will be analyzed by using Fourier Transformed Infrared
Spectroscopy. After that, the sample undergo extraction process under few
parameters. Lastly, the essential oil collected is analyzed with Fourier
Transformed Infrared Spectroscopy.

3.2 Materials, Chemical, Equipment

In this study, several …………………………………………………………….

as despicted in table……

Material Apparatus Equipment

Lemongrass beaker FTIR
ethanol Oven
Ultrasonic bath

25
3.4 Outline Procedures

Citral oil analysis??

Figure 3 1 Overall Flow Chart

26
3.5 Experimental Work

3.5.1 Sample Preparation

The lemongrass leave was obtained from………The lemongrass obtained

are chopped and with a sharp knife. The lemongrass is then washed with tap
water until clean from any dust and were dried in a drying oven at 60°C for 24
hours. Dried lemongrass was separated into two parts that is to be chopped to
4-5cm for chunk size and another part grinded roughly in a dry blender until 2-
3cm for grind size. The dried lemongrass was also analyzed its chemical
composition using Fourier Transformed Infrared Spectroscopy.

Figure 3.1 Dried Lemongrass

27
3.5.2 Extraction Process

The extraction process was done by using the ultrasonic assisted

extraction. This method was carried by using an ultrasonic bath with the settings
for temperature. The experiment ran under different parameters which are size,
time, sample to solvent ratio.

Figure 3.2 Ultrasonic Cleaner

The dried lemongrass was weighted for 15g by using analytical balance
and was put in a 250ml beaker. 100mL of ethanol solvent was measured with
measuring cylinder and transferred into the beaker for extraction. The beaker
with content is then covered with aluminium foil and immersed inside an
ultrasonic bath filled with tap water. The experiment was carried under these
parameters: size of raw material (chunk and grind), time of extraction (2, 3, 4
hours) and sample to solvent ratio (1:1, 1:2, 1:3) that is a total of 12 runs. The
temperature of bath used was 70°C.

28
 Figure 3.3 Extraction Process

After the extraction process, the content of beaker was sieved with a
sieve and the essential oil was transferred into a test tube. The essential oil is
then sent into the rotary evaporator at 78°C that is ethanol evaporating point to
separate the ethanol content leaving only the extract. The extract obtained was
measured and results are recorded.

Figure 3.4 Rotary Evaporator

29
3.5.3 Analysis of Sample

3.5.3.1 Fourier Transformed Infrared Spectroscopy

The analysis was carried by using Fourier Transformed Infrared

Spectroscopy. The essential oil collected are transferred into agate mortar
containing KBr powder. The content was grinded using agate pestle until fine
and mixed completely. The die is assembled with lower pallet polished face up
and the ground mixture is transferred into a cylinder bore and distributed evenly.
The pellet is then closed with another pellet and pressed in a hydraulic press
forming a pellet. The die is removed from the press and sample is analyzed
inside the Fourier Transformed Infrared Spectroscopy machine. Operating
condition for FTIR

Figure 3.5 KBr Powder and Lemongrass Extract Grinded

30
 CHAPTER 4: RESULT AND DISCUSSION

4.1 Introduction
In this chapter, the results of the extraction process run by the ultrasonic
assisted extraction method will be explain and discussed. The theory and cause
of every outcome obtained under the parameter size of raw material (chunk and
grind), time of extraction (2, 3, 4 hours) and sample to solvent ratio (1:1, 1:2,
1:3) will be elaborate briefly. Analysis results of Fourier Transformed Infrared
Spectroscopy for the raw material and essential oil will be compared and
discussed detailly in this chapter of study.

4.2 Experimental Design

To extract the essential oil from the raw material, lemongrass ultrasonic
assisted extraction process was used. There are three different parameters for
this experiment that is size of raw material (chunk and grind), time of extraction
(2, 3, 4 hours) and sample to solvent ratio (1:1, 1:2, 1:3). All these parameters
are made to investigate the effects of different parameters on the oil yield of
extraction process. The analysis process was carried out by using Fourier
Transformed Infrared Spectroscopy to analyze the chemical composition of the
raw material and extract obtained.

31
4.3 Ultrasonic Assisted Extraction
Ultrasonic assisted extraction was used to extract essential oil from the
lemongrass. In this extraction process, 6 samples each for the size of material
chunk and grind was run under different parameters which is time of extraction
(2,3,4 hours) and sample to solvent ratio (1:1, 1:2, 1:3) by using the ultrasonic
bath. The duration for sample to solvent ratio is 2 hours and the temperature of
water bath is 70°C. The volume for 1:1 solvent is 100ml of ethanol.

Figure 4 1 Ultrasonic Bath Cleaner

32
4.3.1 Time of Extraction
Time of Volume of Extracted Percentage Oil
Samples Size Extraction Essential Oil (ml) Yield (%)
(Hours)
1 2 2.0 2.00
2 3 3.6 3.60
Chunk
3 4 5.4 5.40
4 2 4.5 4.50
5 3 5.4 5.40
Grind
6 4 6.2 6.20
Table 4 1 Result for Extract with Time Parameter

From Table 4 1, sample 1, 2, 3 is for the size of sample chunk while

sample 4,5,6 is for the size grind. These samples were extracted using
ultrasonic assisted extraction with the solvent ethanol 100ml per run. From the
result recorded, the volume essential oil extracted from sample 1 is 2.0 ml, for
sample 2 is 3.6 ml, for sample 3 is 5.4 ml that is for the time of extraction 2, 3, 4
hours with the size of raw material chunk. The oil yield percentage are 2.0, 3.6
and 5.4% respectively. For the size of raw material grind, sample 4 obtained 4.5
ml, sample 5 is 5.4 ml and sample 6 is 6.2 ml. The percentage of oil yield are
4.50, 5.40, and 6.20% respectively. At the next page, Figure 4 2 and 4 3 shows
the graph for the Extract Volume (ml) vs Time of Extraction (Hours) and Oil Yield
Percentage vs Time of Extraction (Hours) for both chunk and grind size of raw
material.

33
 Extract Volume (ml) vs Time of Extraction (Hours)
7.00
6.00

Extract Volume (ml)

f(x) = 0.85 x + 2.82
5.00 f(x) = 1.7 x − 1.43 Chunk
4.00 Linear (Chunk)
3.00 Grind
2.00 Linear (Grind)
1.00
0.00
1.5 2 2.5 3 3.5 4 4.5
Time (hr))

Figure 4 2 Extract Volume (ml) against Time of Extraction (Hours)

Oil Yield Percentage (%) vs Time of Extraction (Hours)

7.00
Oil Yield Percentage (%)

6.00
f(x) = 0.85 x + 2.82
5.00 f(x) = 1.7 x − 1.43 Chunk
4.00 Linear (Chunk)
3.00 Grind
2.00 Linear (Grind)
1.00
0.00
1.5 2 2.5 3 3.5 4 4.5
Time of Extraction (Hours)

Figure 4 3 Oil Yield Percentage (%) against Time of Extraction (Hours)

From Figure 4 2, the extract volume for both size of material chunk and
grind increase over the time of extraction. The difference is that the extract
volume for grind size is higher than the size of chunk. Figure 4 3 shows the oil
yield percentage (%) increases over time similar with Figure 4 2. The highest
difference between size chunk and grind is at time of extraction 2 hours. Size of
material chunk obtained 2.0 ml extract volume, on the other hand; size of raw
material grind collected more than 50% of the volume extract for chunk that is
4.5ml. Fully observed, both size increases oil yield over time but grind size has
the higher efficiency.

34
4.3.2 Sample to Solvent Ratio
Samples Size Sample to Solvent Volume of Extracted Percentage Oil Yield
Ratio Essential Oil (ml) (%)
1 1:1 1.8 1.80
2 1:2 4.8 2.40
Chunk
3 1:3 7.6 2.50
4 1:1 4.7 4.70
5 1:2 6.4 3.20
Grind
6 1:3 8.7 2.90
Table 4 2 Result for Extract with Sample to Solvent Ratio

From Table 4 2, sample 1, 2, 3 is for the size of raw material chunk while
sample 4, 5, 6 is for the size of raw material grind. The parameter used are
different sample to solvent ratio that is 1:1, 1:2, and 1:3. For sample 1, extract
collected is 1.8ml, for sample 2 is 4.8ml and for sample 3 is 7.6ml. The
percentage oil yield is 1.80, 2.40, and 2.50% respectively. For the size of raw
material grind, sample 4 obtained 4.7ml extract, sample 5 is 6.4ml and sample 6
is 8.7ml. The oil yield percentage is 4.70, 3.20, and 2.90% respectively. Figure 4
4 and 4 5 at the next page shows the graph for Extract Volume (ml) vs Sample
to Solvent Ratio and Oil Yield Percentage (%) vs Sample to Solvent Ratio.

35
 Extract Volume (ml) vs Sample to Solvent Ratio
10.00

Extract Volume (ml)

8.00 f(x) = 2 x + 2.6
f(x) = 2.9 x − 1.07 Chunk
6.00 Linear (Chunk)
4.00 Grind
2.00 Linear (Grind)
0.00
1:1 1:2 1:3
Sample to Solvent Ratio

Figure 4 4 Extract Volume (ml) against Sample to Solvent Ratio

Oil Yield Percentage (%) vs Sample to Solvent Ratio

5.0
Oil Yield Percentage (%)

4.0 f(x) = − 0.9 x + 5.4

Chunk
3.0
Linear (Chunk)
2.0 f(x) = 0.35 x + 1.53 Grind
Linear (Grind)
1.0
0.0
1:1 1:2 1:3
Sample to Solvent Ratio

Figure 4 5 Oil Yield Percentage (%) against Sample to Solvent Ratio

From Figure 4 4, the extract volume for both chunk and grind size
increased over sample to solvent ratio increase. The data shows that grind size
has higher extract volume over the size chunk. Figure 4 5 shows the oil yield
percentage of both size over the sample to solvent ratio. From the data, it can
be observed that the percentage of oil yield for the size grind declining over the
sample to solvent ratio while the trendline for the size chunk increases steadily.
Nevertheless, the oil yield percentage for the size grind never goes under the
percentage of oil yield for the size chunk. Inspecting both data, the oil yield for
each size increases just for the grind size, increasing of solvent volume has a
lower efficiency.

36
 Theoretically, size of raw material plays a big role on the yield obtained
during the extraction. Smaller size of raw material enables more area of effect
for the solvent (ethanol) to conduct composition transfer [ CITATION Sai14 \l 1033 ] .
Other than that, ultrasonic waves can penetrate the cell wall efficiently due to the
small particles. Rapid extraction and solvent diffusivity can be elevated better if
the particle is ground fine. On the other hand, bigger size of raw material leads
to slow extraction process. The total area exposed is smaller than the small
particles. Moreover, it takes a few times for the ultrasonic waves to penetrate all
the cell wall of the lemongrass thus slowing down composition transfer [ CITATION
Kha11 \l 1033 ].

Referring Figure 4 2 and 4 3, the oil yield obtained increases over time.
Going up the time 2, 3, and 4 hours, the extract volume collected are different
and increasing as the time increases. The longer the time of extraction, more
glandular walls are ruptured, and solvent diffusion can operate mass transfer
efficiently [ CITATION Mar13 \l 1033 ]. Warning ahead, exposal of heat for a long
time may cause damage to the extract such as thermal degradation. The
structure of essential oil collected can be affected as they are prone to damage
such as oxidative damage, chemical transformation, or unnecessary
polymerization [ CITATION Sti13 \l 1033 ]. From Figure 4 2, it can be observed the oil
yield for the size chunk increases from 2.0 ml at 2 hours to 3.6 ml at 3 hours and
5.4 ml at 4 hours. At Figure 4 2 also can be observed the increase of oil yield for
the size grind from 4.5 ml at 2 hours to 5.4 ml at 3 hours and at 4 hours it
increases to 6.2 ml. All these occur under the same temperature (70°C) and
same solvent volume (100 ml of ethanol) by changing only its time of extraction.

37
 Based on Figure 4 4 and 4 5, the oil yield recorded increases over the
sample to solvent ratio increase. The usual solvent volume used is 100ml and
the sample to solvent ratio tested are 1:1, 1:2, and 1:3. Looking into the extract
volume collected for raw material size chunk are 1.8 ml for 1:1 ratio, 4.8 ml for
1:2 ratio, and 7.6 ml for 7:6 ratio. Referring Figure 4 5, the percentage of oil yield
seems increasing over the increase of solvent used. For grind size raw material,
the extract volume produced are, 4.7 ml for 1:1 ratio, 6.4 ml for 1:2 ratio and at
1:3 ratio the extract volume is 8.7 ml. However, the percentage oil yield has
been seen at declining rate. The percentage falls from 4.7% at 1:1 ratio to 3.2%
at 1:2 ratio and 2.9% at 1:3 ratio. According to a research, the declining rate of
percentage happened due to too much solvent used. Too much solvent leads to
the stop of extraction process because of the solvent diffusivity making the
material and solvent reach an equilibrium point thus mass transfer cannot be
operate anymore [ CITATION Ach04 \l 1033 ] . Therefore, increase of solvent at small
size raw material is not efficient and the oil yield for chunk size raw material can
be predicted to be obtained more than grind size if the time of extraction is
extended [ CITATION Sai14 \l 1033 ].

4.4 Fourier Transformed Infrared Spectroscopy

Fourier Transformed Infrared (FTIR) Spectroscopy was employed to
characterize the different functional group of a material. The spectrum recorded
use a range of 4000-400cm -1. Dried lemongrass and lemongrass essential oil
obtained undergo FTIR analysis to detail its components and changes on the
raw material and the extract. Figure 4 6 and Figure 4 7 are the results of FTIR
analysis to dried lemongrass and lemongrass extract. In this analysis, hydroxyl
ion (OH-) and carbonyl group (C=O) are expected as Citral (C 10H16O) is a
component makes up from these molecules.

38
 Figure 4 6 Dried Lemongrass FTIR Result

Figure 4 7 Lemongrass Extract FTIR Result

Lemongrass is a citral plant which its chemical compound is C 10H16O.
Based on this, it is to be expected that there is hydroxyl ion (OH -) and carbonyl
group (C=O) when it is analyzed [ CITATION Dhe15 \l 1033 ]. Referring the IR
Spectrum Table for frequency range, hydroxyl ion (OH -) ranges from 3000-
4000cm-1 and carbonyl group (C=O) ranges from 1600-1700cm -1 [ CITATION
IRS20 \l 1033 ]. From Figure 4 6 and 4 7, it can be observed that there is a peak at
3446.56 and 3409.44cm-1, respectively. This phenomenon shows the existence
of hydroxyl ion (OH-) where the range is from 3000-4000cm -1. Next, is there is
also a peak can be observed at wavelength 1638.16 and 1637.58cm -1 at the
FTIR result. Looking back at the IR table, carbonyl group (C=O) ranges from

39
1600-1700cm-1 thus it is believable that carbonyl group is detected in these
compounds. Therefore, citral existence is confirmed in the compound of both
dried lemongrass and lemongrass extract. Other than that, lemongrass also
contains terpenes such as citronellal, nerol, limonene and geraniol [ CITATION
Lem18 \l 1033 ]. Therefore, the peak at around 2900cm -1 stands for C-H, C=O and
O-H peak at around 1700cm-1 and 3400cm-1 as mentioned before, and C-O
peak at around 1100cm-1 confirms there is terpenes is one of lemongrass’
compositions.

40