ACTIVATING EFFECT OF AROMATHERAPY

INHALATION WITH Citrus hystrix (KAFFIR

LIME) LEAVES OIL

ALISA STEPHANIE RASION

11". sfr«.. .... : ¢ý'si: lJ1ii Liý/: r. `t

, ýi

THESIS SUBMITTED IN FULFILLMENT FOR

THE DEGREE OF MASTER OF SCIENCE

FACULTY OF SCIENCE AND NATURAL

RESOURCES UNIVERSITY OF MALAYSIA

SABAH 2018

UNIVERSITI MALAYSIA SABAH

BORANG PENGESAHAN STATUS TESIS

JUDUL: ACTIVATING EFFECT OF AROMATHERAPY INHALATION WITH Citrus hystrix (KAFFIR LIME) LEAVES OIL

IJAZAH: SARJANA SAINS (KIMIA INDUSTRI)

Saya ALISA STEPHANIE RASION, sesi 2016-2018, mengaku membenarkan tesis Sarjana ini disimpan di Perpustakaan Universiti Malaysia Sabah dengan syarat-syarat kegunaan seperti berikut: - "

1. Tesis ini adalah hak milik Universiti Malaysia Sabah 2. Perpustakaan Universiti Malaysia Sabah dibenarkan membuat salinan untuk tujuan

pengajian sahaja. 3. Perpustakaan dibenarkan membuat salinan tesis ini sebagai bahan pertukaran antara

institusi pengajian tinggi. 4. Sila tandakan (/):

x

üýtºi6r: iýi:: 11 f Hi1ýc11 Uia VIYL, 'ýi ý

SULIT (Mengandungi maklumat yang berdarjah keselamatan atau kepentingan Malaysia seperti yang termaktub di dalam AKTA RAHSIA 1972)

TERHAD (Mengandungi maklumat TERHAD yang telah ditentukan oleh organisasi/badan di mana penyelidikan dijalankan

TIDAK TERHAD

Disahkan Oleh,

ALISA STEPHANIE RASION MS1521001T

NURULAIN BINTI ISMAIL PUSTAKAWAIWANAN

ABAH

(Tandatangan Pustakawan)

Tarikh :V October 2018 (Prof. Madya. Dr. How Siew Eng) Penyelia

DECLARATION

I declare that this thesis and the work presented in it are my own and has been

generated by me as the result of my own original research.

27t'' September 2018

Alisa Sianie Rasion MS1521001T

11

CERTIFICATION

NAME ALISA STEPHANIE RASION MATRIC NO. MS1521001T

TITLE ACTIVATING EFFECT OF AROMATHERAPY

INHALATION WITH Citrus hystrix (KAFFIR

LIME) LEAVES OIL DEGREE : MASTER OF SCIENCE (INDUSTRIAL

CHEMISTRY)

VIVA DATE 9TH AUGUST 2018

CERTIFIED BY;

SUPERVISOR

PROF. MADYA. DR. HOW SIEW ENG

SIGNATURE

"nýý ö

ACKNOWLEDGEMENTS

First and foremost, praises and thanks to the God, the Almighty, for His showers of blessings throughout my research work to complete the research successfully. I

would like to express my sincere gratitude to my advisor Prof. Madya. Dr. How Siew Eng for the continuous support of my master study and research, for her patience, motivation, and immense knowledge. Her guidance helped me in all the time of research and writing of this thesis and finally I am extremely grateful to my parents and to my friends for providing me with unfailing support and continuous encouragement throughout my years of study and through the process of researching and writing this thesis. This accomplishment would not have been

possible without them. Thank you.

Alisa Stephanie Rasion

27th September 2018

iv

ABSTRACT

Citrus hystrix leaves oil has a scent with unique nuances because it is rich in natural constituent citronellal which possesses soothing properties and has the potential to be used as aromatherapy. However, there have been relatively few published about the C. hystrix leaves extract and its efficacy as aromatherapy. This research was aimed to investigate the effects of C. hystrix leaves oil on human behavioural parameters after inhalation. C. hystrix essential oil was obtained from fresh leaves by steam distillation and the oil yield obtained was 0.14% with pale yellow in colour while the chemical profile of C. hystrix leaves volatile oil was analysed using the gas cromatography-mass spectrometry (GC-MS). It was found that the citronellal is the highest percentage (85.00%) followed by citronellol (4.75%), citronellyl propanoate (2.74%), delta-3-carene (1.15%), geranyl acetate (0.84%), (E)-ß-ocimene (0.69%), linalool (0.42%), terpinolene (0.40%), myrcene (0.35%), camphene (0.25%), ß- pinene (0.24%), and a-Phellandrene is the lowest percentage (0.05%). 3-(4,5- dimethylthiazol- 2yl)-2,5-Diphenyl tetrazolium bromide (MTT) test was used to measure cytotoxicity of the oil and from this research it was found that the essential oil of C. hystrix (leaves) showed less toxic to the normal human liver cells (WRL 68) with the highest value of ICso (40.23±14.86 ug/mL) where it required a higher concentration of the C. hystrix (leaves) oil to inhibit the activity of the cells by 50% while the low dose concentration of paclitaxel reduced the percentage viable cells (0.0016±0.0003 ug/mL). There were 40 respondents participated this survey which were randomly assigned into two groups, control group (treated with virgin coconut oil (VCO) since this oil has no scent and it is typically used as a carrier oil) and the C. hystrix oil group and each group consists of 20 respondents. Visual analog scale (VAS) was used for this survey for the subjects to rate their emotional condition in order to assess subjective behavioural arousal in terms of relaxation, vigour, calmness, attentiveness, mood and alertness after inhalation. Data obtained in this study were analyzed using Mann-Whitney-U-Test, Wilcoxon Signed Rank Test, and Spearman's Rho Correlation. Based on the Mann-whitney U Test result, it showed that C. hystrix oil gave arousal effects in relaxation, vigorous and attentiveness and this data was supported by Wilcoxon signed rank test data for beginning and ending time where the data of C. hystrix oil category were significantly different in score of effects at the beginning and ending observation data for relaxation, vigorous and attentiveness rated by respondents. From this finding, C. hystrix oil gave arousal effects in relaxation, vigorous and attentiveness and these were supported by the Spearman's Rho Correlation test where relaxation had positive medium correlation with variable vigorous, r=0.47, n= 20, p<0.05, and variable vigorous had positive strong correlation with variable attentiveness, r=0.74, n= 20, p<0.05. The C. hystrix leaves oil has been experimentally shown to have a positive effect on relaxation, vigorous and attentiveness than subjects in the control group and these results provide evidence for its use in aromatherapy for the relief of depression and uplifting mood in humans and this might be due to higher content of citronellal in C. hystrix leaves oil and it is safe to be used.

V

melegakan kemurungan dan menaikkan mood manusia dan ini mungkin disebabkan o%h kandungan citronellal yang tinggi di dalam daun minyak pati C. hystrix dan is selamat digunakan.

vii

TABLE OF CONTENTS

TITLE

DECLARATION

CERTIFICATION

ACKNOWLEDGEMENTS

ABSTRACT

ASSRRAK

TABLE OF CONTENTS

LIST OF TABLES

LIST OF FIGURES

LIST OF ABBREVIATIONS/ SYMBOLS

LIST OF APPENDICES

CHAPTER 1: INTRODUCTION

1.1 Introduction

1.2 Significance of the Study

1.3 Scope of the Study

1.4 Research Objectives

CHAPTER 2: LITERATURE REVIEWS

2.1 Aromatherapy

2.2 Essential Oil

2.3 Composition of Essential Oil 2.4 Actions of Essential Oil On the Body

2.5 Aroma-therapeutic Effect of Essential Oil on Human Behavioural and Emotional Changes

2.6 Olfaction Physiology 2.7 Psychological Evaluation of Fragrance Effects Using a Visual Analog

Scale (VAS)

Page

i 11

III iv

v VI

viii X

XII

XIII

xv

1

1 4

5

5

6

6

8

11

12

14

15

17

viii

2.8 Genus Citrus and Its Therapeutic Benefits 18

2.9 Kaffir Lime 21

2.10 Steam Distillation 26

2.11 Citronellal 27

2.11.1 Source of Citronellal 28

2.11.2 Biological Benefits of Citronellal 29

2.11.3 Toxicity of Citronellal 33

2.11.4 Method to Analyse the Toxicity of Citronellal 35

CHAPTER 3: RESEARCH METHODOLOGY 37

3.1 Sample Preparation 37

3.2 Extraction Method 38

3.3 Analysis of Volatile Compound 39

3.4 Toxicity Test 40

3.5 Subjects 42

3.6 Data Analysis 43

CHAPTER 4: RESULT AND DISCUSSION 45

4.1 Chemical Component of C. hystrix Volatile Oil 45

4.2 Toxicity of C. hystrix Oil 53

4.3 The Stimulating Effect of C. hystrix(Leaves) Oil Aroma on Human 54

4.3.1 Comparison of Control and Treatment Related to Six Variables 55

4.3.2 Comparison Beginning and Ending 56 4.3.3 Relationship Among Variables 59

CHAPTER 5: CONCLUSION

5.1 Conclusion 5.2 Future Research

REFERENCES

APPENDICES

63

63

64

65

81

ix

LIST OF TABLES

Page

Table 2.2 Percentage composition of leaves 22

oils of Kaffir lime

Table 3.1 GC-MS conditions for extracted oil 39

analysis

Table 3.2 Definition of relaxation, vigorous,

calmness, attentiveness, mood and

alertness

43

Table 4.1 Chemical composition of C. hystrix 45

leaves oil using GC-MS

Table 4.2 Comparison of chemical

composition of Citrus hystrix leaves

oil between the present data and

previous data of the existing literature (%)

Table 4.3 In vitro cytotoxicity of essential oil

of C. hystrix (leaves)

46

54

Table 4.4 Comparison of mean rank and sum 55

of rank between control and C. hystrix leaves oil category for the

effects of relaxation, vigorous,

calmness, attentiveness, mood and

alertness

X

Table 4.5 Rating mean (±standard deviation) 57

and comparison of rating at the beginning and ending observation time of six variables rated by 20

respondents for the effects of Control

Table 4.6 Rating mean (±standard deviation) 57

and comparison of rating at the beginning and ending observation time of six variables rated by 20

respondents for the effects of Citrus hytrix leaves oil

Table 4.7 Spearman's Rho Correlation

between six variables for the effects

of Citrus hytrix leaves oil

60

XI

LIST OF FIGURES

Page

Figure 2.1 The flow of air inhalation 13

into the nostrils

Figure 2.2 Structure of the

Citronellal

Figure 2.3 Structure of the

Citronellol, Citronellal and Hydroxycitronellal

27

29

Figure 3.1 Kaffir lime leaves 37

Figure 3.2 Steam distillation (40L) 38

Figure 4.1 The chromatogram of C. 49

hystrix oil components

Figure 4.2 The chromatogram of 50

standard citronella)

Figure 4.3 The chromatogram of C. 51 hystrix oil confirmed with the citronellal

XII

LIST OF ABBREVIATIONS/ SYMBOLS

Rpm

9

Kg

L

mL

Nm

mg

m

mm

eV

N9

NL

%

oc

Revolutions per minute

Gram

Kilogram

Liter

Mililiter

Micrometer

Miligram

Meter

Milimeter

Electronvolt

Microgram

Microlitre

Percentage

Degree Celsius

CO2 Carbon dioxide

XIII

nm

WRL68

DMEM

PSG

FBS

NMDA

ICso

L-NAM E

Ca2+

f

>

>

Nanometre

The human hepatic cell line

Dulbecco's Modified Eagle Medium

Penicillin-Streptomycin-Glutamine

Fetal bovine serum

N-methyl-D-aspartate (receptor that is used to control the synaptic

plasticity and memory function)

Inhibitory concentration by 50%

N(uo)-nitro-L-arginine methyl ester (induced hypertension)

Calcium ions

Standard deviation

Greater than or equal to

Less than

Greater than

XIV

LIST OF APPENDICES

Page

Appendix A The mass spectrum of each of the 81 identified compounds in C. hystrix leaves oil

Appendix B Library of GCMS 88

Appendix C GCMS (C, hystrixoil components) 90

Appendix D GCMS (C. hystrixoil confirmed 91 with the citronellal)

Appendix E Visual Analog Scale for the effect 92

of aromatherapy assessment

Appendix F Rating (Meant SD) for the

stimulating effect of control and C. hystrix aroma at the beginning

and ending observation time

Appendix G Research permission letter

Appendix H Consent form

99

111

113

xv

CHAPTER 1

INTRODUCTION

1.1 Introduction

Aromatherapy is defined by Sense of Smell Institute (SSI) as the therapeutic effects

of aromas on physical conditions such as digestive problems and menstrual disorders

as well as psychological conditions such as chronic depression (Balchin, 2006). In a

modern aromatherapy, both essential oils and their aromas are used for balancing,

harmonizing and promoting the health of body, mind and spirit where the effects of

an aroma can be instantaneous including both direct and indirect psychological

effects even thinking about a smell may have a similar effect to the smell itself.

However, some evidences showed that inhaled or dermal applied essential oils enter

the blood stream and in relevant molecular, cellular or animal models, exert

measurable psychological effects, indicates that the effects are primarily cabenefits.

Extraction obtained from mostly basic plant parts such as stems, seeds, flowers, resins, leaves, barks, roots, or fruit rinds produced essential oils which are highly concentrated substances (Rao and Pandey, 2006). Essential oils flavour and their therapeutic or odoriferous properties often used in foods, medicines, and

cosmetics. There are three ways essential oils can be absorbed including through the

olfactory and respiratory systems (vapour inhalation), transdermally through lotions

or compresses involving massage and during bathing, or orally, through consumption

of essential oils in capsules or as additives to food or medicinal preparations, for

example but it is more to the herbal medicine than aromatherapy (Perry, and Perry, 2006). Citrus essential oil is the good source of aromatherapy and it was the most popular source of perfume and fragrance essences because of its good sour and sweet tastes, pleasant and refreshing aroma, as well as extensive growing areas worldwide.

1

There are several plants of Citrus rich in essential oil often used in the anxiety

treatment and among these plants are orange, Citrus sinensis (Faturi et al., 2010),

Citrus aurantium, bergamot (Carvalho-Freitas and Costa, 2002; Pultrini etat., 2006),

Citrus Amon, lemon (Ceccarellia et al., 2004) and these essential oils are often used

as aromatherapy agents for relieving and for reducing stress and depression which

have been used topically for their antibiotic and antiviral properties for over the years

(Setzer, 2009; Cooke and Ernst, 2000). Since the genus Citrus essential oil showed

positive emotional effects in studies with humans and demonstrated anxiolytic as well

as sedative effects in experimental models of anxiety and hence, it could be a great

alternative (Pimenta et al., 2012). Hence, C. hystrix(Kaffir lime) leaves will be used

for this research which is from the same genus Citrus and the C. hystrix leaves that

have a strong fragrant lemon scented. It was previously reported that heart disease,

dizziness and indigestion, as well as physical nourishment, could be treated by C.

hystrix which used as herbal medicine (Abdul, 2013). C. hystrix leaves have

antimicrobial properties and commonly been used for cooking and its essential oil or

oleoresin for aromatherapy (Sid et al., 2016).

C. hystrix is an indigenous plant in Southeast Asia including Malaysia,

Indonesia and Thailand. C. hystrix from Rutaceae family has it's own aroma and

commonly used as spices in cooking and medicinal herbs. C. hystrix has a bumpy

fruit with a rough skin while the leaves are an aromatic herb which used broadly in

adding a distinctive aroma and flavour to food. There are two types of essential oil

made from C. hystrix, one from the peel and the other from leaves. The C. hystrix

leaves contain oleoresin such as citronellal which give it aroma. The major component

in the C. hystrixleaves is citronellal (Kawiji, eta/., 2015). Basically, the higher content

of geraniol, citronellal hydrate, citronellal, linalool, linalyl acetate, the more delicate

its aroma (Kawiji et al., 2015). The major component of the ethyl acetate extract from leaves of C. hystrixis citronellal (73.08%) and that from hydrodistillation is also

citronellal (100%) while the major components of the hydrodistillated essential oils from C. hystrix peels are limonene (16.85%) and cironellal (3.57%) whereas

citronellal (2.80%), limonene (0.31%), and pinene (0.13%) were major components

of essential oil of the ethyl acetate extract (Megawati, 2009). Exposure to C. hystrix

2

aroma produced the effect of reducing "disguise iritation" which is evaluated based

on cognitive evaluation and it indicates that exposure to the C. hystrix aroma

produces a positive emotive response which may impact clarity of mind and health

(Megawati, 2009). C. hystrix could be a great alternative and has the potential in

research and commercialization for the solution in insect repellent, aromatherapy,

and spa practices, making shampoo, beauty product as well as antioxidants

compound (Nurhani etal., 2013).

Since Citrus hystrix leaves oil has a scent with unique nuances because it is

rich in natural constituent citronella) which possesses soothing properties and has

the potential to be used as aromatherapy. Steam distillation was used in this research

because steam distillation produces more citronellal from the leaves than the other

methods where previous results reported that citronellal was the major component

in C. hystrix leaves oil with the highest percentage (80.04%) (Srisukh et al., 2012)

compared to other techniques such as solvent extraction, hydrodistillation,

supercritical fluid extraction, pressurised liquid extraction (PLE), and maceration

extraction where the results report that the percentage of citronellal is 74.8%,

66.85%, 1.41%, 25.87%, and 25.66% respectively (Tinjan and Jirapakkul, 2007; Loh

etal., 2013; Haiyee and Winitkitcharoen, 2012; Kawiji etal., 2015).

Citronellal gives therapeutic benefits including antiviral, antimicrobial, tonic,

vasodilators, a hypotensive, calming, antipyretic, sedative, and spasmolytic (Bouayed

and Bohn, 2012) and at the various activity levels, C. hystrix leaves oil has excellent

activity against many respiratory bacteria (Srisukh etal., 2012). Thus, it can be used

as air purifying to clean the air naturally using the air purifier. Steam distillation is

used to extract essential oil for this research because it is economy for small and

medium scale industry, cheap for a basic level process, as well as there is no changing

properties of oils produced by this method. The oil never spoil in this method since the steam reduces the boiling point of a particular component of the oil and this

method also faster than other methods (Kumar, 2010). Steam distillation is the oldest

3

method and simplest method to produce essential oil using leafy materials for

commercial purpose.

Before the essential oil of C. hystrix can be tested on humans, toxicity test

should be conducted first. Safety tests begin with acute toxicity testing where a single

dose of the test compound is given to animals and this test is focusing on determining

the range between the two possible doses which is no adverse effect and the dose

that is life-threatening. Toxicity divided into two types which are acute or chronic and

these types are based on the number of exposures to a poison as well as the time it

takes for toxic symptoms to develop. Short-term exposure with a high dose causes

acute toxicity while the repeated or long-term exposure with low doses causes

chronic exposure. MTT test is commonly used for cytotoxicity testing because of their

rapid, standardized and sensitive in determining if a material contains significant

quantities of hazard components. The MTT (3- [4,5-di m ethyl th iazol-2-yl]-2,5 diphenyl

tetrazolium bromide) assay determines the cells activity where it is based on the

conversion of MTT into formazan crystals by living cells (Meerloo et al., 2011). MTT

test can be extrapolated the results to toxicity to mammals since it uses tissue culture from cell (Tunjung et a/., 2015). MTT assay is more suitable technique for the

application to toxicity test for human and because for this reason MTT test is used for this research to determine the C. hystrix oil toxicity.

1.2 Significance of the Study

Hectic daily tasks of modern life can give a stressful life and people nowadays tend to take drugs of pain killer to stop the pain or the uncomfortable symptom that linked

to depression, anxiety and stress but drugs are toxic to the body and have the side effects compare to the natural molecules such as those found in essential oils. An

essential oil is multi-dimensional and filled with homeostatic intelligence where it has the capability in restoring the body to healthy balance and if there are changes in body conditions, the oil will adapting, increasing or reducing blood pressure as needed, stimulating or suppressing enzyme activity as needed, energizing or relaxing

4

as needed (Stewart, 2005). Treatment with essential oils is safer and fewer adverse

effects.

1.3 Scope of the Study

There have been relatively few published about the C. hystrix leaves oil and its

efficacy as aromatherapy. Therefore, the present study of C. hystrix leaves oil is to

investigate the potential of inhalation with C. hystrix leaves oil as aromatherapy on

healthy human emotional responses where the scope of this research is to study the

positive effect of C. hystrix leaves oil on stimulation of human relaxation, vigour,

calmness, attentiveness, mood and alertness which linked to individual performance.

Stimulating oil gives expected effect on mind and body. The benefits of this research for users are to improve the emotional balance and to improve overall attitude which

increases the life quality and health. Users can effectively perform their tasks and

make a maximum efficiency.

Research Objectives

Objectives of this research are:

a) To determine the chemical components of C. hystrix oil using Gas

cromatography-mass spectrometry. b) To evaluate the toxicity of C. hystrix oil.

c) To evaluate the stimulating effect of C. hystrix aroma on humans such as

relaxation, vigour, calmness, attentiveness, mood and alertness.

5

CHAPTER 2

LITERATURE REVIEW

2.1 Aromatherapy

Aromatherapy is practicing the use of natural oils and which also referring to essential

oils therapy. In aromatherapy, the main therapeutic agents are essential oils which

are highly concentrated substances extracted from plants and these oils are good to

apply in the treatment of mental and physical balance which give stress relief, revive

and revitalize the individual for a next day's duty (Ali et al., 2015). Aromatherapy is

the holistic practice of preventative approach and as an active technique to use during the stages of acute and chronic illness. The use of aromatic plants has been

around since Neolithic times and these aromatic plants have been used for beauty

treatment, perfuming, medicinal practices, cleansing, and ritual bathing as well as

religious ceremonies (Stiles, 2017).

In fragrance psychology field, there is a various number of group for study including perfumes (e. g. colognes, fine fragrances, ), cosmetic and toiletry products

with scent (e. g. hair-care, fragranced skin-care, and hand care products), environmental fragrances (e. g. room fresheners, potpourri) as well as aromatherapy

products (essential oils) (Graham, 2000). Fragrance psychology is the perception of individual odour where these odours are likely to affect a various field of human

psychological functioning, ranging from perception and mood to cognitive processes and attitude as well as the changes in humans induced by smelling the fragrances of essential oils (Lenochova et at., 2012; Sugawara et at., 2013). The psychology includes the product behavioural effects on the user including others perceptions (personality, the appearance, and projected image) self-perception (moods, self- image, confidence, self-esteem, and attitudes), interpersonal attraction (sexual

attraction and liking) and in psychology field also includes motivations or reasons for

using or purchasing a product which relating scent to differentiate situation and

6

lifestyles and to link individual fragrances with particular personality aspect and finally

it access therapeutic effects including mental and physical particularly to relevant

aromatherapy products (Graham, 2000).

Application of single or combined essential oils including diffusing the oils into

inhaled air, using in massage oil or adding to bathwater. The inhalation of these

essential oils molecules triggers the olfactory nerve which sending the messages to

the brain's limbic system including the cognitive, memory, and emotion that are said to stimulate the physiological responses for examples lavender promotes relaxation and eucalyptus relieves congestion. The utilization of extracted aromatic plant essence gives the aromatherapy benefits where the treatment outcome could be

achieved by conveying the odours to the central nervous system via the skin and olfactory system. The odours are dispersed in the blood and circulate to different

organs which can prevent and treat the central nervous system diseases and for this

reason, this application makes the aromatherapy as compatible therapies where the

essential oils as the major therapeutic agents can treat several diseases (Lee et al., 2011).

The major methods used in aromatherapy that use the essential oils are including inhalation, local application, and baths. These oils penetrate the human

skin surface and once the oils are in the system, they regulate themselves and fix the affected area or malfunction site and this type of therapy uses various permutation and combinations to get relief from numerous illness such as depression, insomnia, headache, indigestion, muscular pain, skin ailments, respiratory problems, urine associated complications, swollen joints and so on (Ali et al., 2015). Aromatherapy can be classified as massage aromatherapy, olfactory aromatherapy cosmetic aromatherapy, medical aromatherapy, and psycho-aromatherapy.

7

2.2 Essential Oil

Essential oils can be defined as volatile substances that are more or less volatile with

more or less odorous impact, which obtained by steam distillation or dry distillation

or by several mechanical treatments. Essential oils are colourless but have a pleasant

smelling liquids with high refractive index. The frequency of essential oils ranges are

several times greater than the frequency of herbs and food and thus when apply the

essential oils, the body's overall frequency increase quickly and stabilize the

struggling organ frequency. Since essential oil is a very concentrated form that

possesses nutritive values and healing properties, and thus the use of this oil can

improve overall immune function by healing the body, mind, and spirit, while reviving

the damaged tissue and bringing oxygen to deprived cells. (Westover, 2012).

Essential oils have the ability to bring the body back to normal ranges within seconds

and improve overall feelings of well-being (Westover, 2012).

Essential oils are commonly used for scenting purposes including food

flavourings and cosmetic fragrances but since aromatherapy is getting popular in

recent years, it also leads to the increase uses of essential oils in aromatherapy.

(Japan external trade organization, 2011). The properties of essential oil such as

antibacterial, anti-infective, antifungal, antiparasitic, anti-tumoral, antiviral, and

antiseptic resulted from its unique oxygen or nutrient carrying capacity of essential

oil and this oil is highly concentrated, low volume and high value products (Joy etal.,

2001). In laboratory studies and clinical trials, essential oils have been shown to

destroy and to inhibit tested bacteria and viruses without causing harm (unlike drugs)

to cells, organs, tissues, or body (Westover, 2012). Most of the medications (drugs)

have potential side effects than the essential oils, for example, morphine which is

the prototype of opioid analgesics can exert side effects including sleepiness,

respiratory depression, motility, endocrine and autonomic nervous systems disorders,

and decreased gastrointestinal nausea (Sarmento-Neto eta/., 2015) while treatment

for superficial and deep infections caused by Trichophyton using Ketoconazole which

used as antifungal drugs give unpleasant side-effects including abdominal pain and

itching, nausea, and the toxicity of these drug limit its therapeutic use in many cases

(Shin and Lim, 2004). Rituximab used for the treatment of low-grade or follicular

8

CD20-positive, Hepatitis B and treatment of rheumatoid arthritis (RA) in combination

with methotrexate. However there is potentially fatal complication in a patient who

recently received rituximab such as caused allergic reaction, blood and lymphatic

system disorders, infectious complications, renal disorders, cardiac disorders,

adverse pulmonary reactions, neoplastic disorders, gastrointestinal disorders,

neurological disorders and developed itching, rash, or urticarial (Kasi eta/., 2012: 4).

The agent to treat the primary posttraumatic stress disorder (PTSD) is

Benzodiazepines but however it is dangerous in over-dosage and often abused

(Bernardy, 2013). Benzodiazepines gave poor sleep overall which leads to increase

the risk of fractures and falls, accidents, and give adverse cognitive effects to the

elderly where a cognitive "slowness" can be difficult to vary from cognitive reduction

(Lader, 2011) as well as causing mild traumatic brain injury (Department of Veterans

Affairs, 2010).

Unlike drugs, the long-term effects of essential oil use are considered mild.

Essential oil use is non-addictive and proven to provide desirable results (Emerald,

2016). Essential oils, such as Lavender (Lavandula angustifolia), Bergamot (Citrus

bergamia) and Ylang Ylang (Cananga odorata), could be implemented into

aromatherapy, holistic and massage practice for the potential management of PTSD

where the daily use of Lavender oil in aromatherapy with PTSD patients help to

decrease depression by 32.7% and to decrease sleep disturbances dramatically,

moodiness and overall ill health in 47 PTSD patients (Uehleke etas., 2012). In multiple

studies, Lavender essential oil has anti-anxiety effects caused by modulation of two

neurotransmitters, 5-hydroxytryptamine and dopamine (Umezu, 2010).

Administration of the Bergamot oil in concentrations of 1.0,2.5, and 5.0% w/w

demonstrates a strong anxiolytic effect measured through different tests, and its

effects on stress-induced plasma corticosterone has been compared to the effects of

diazepam (Saiyudthong and Marsden, 2010). Testing of the Bergamot essential oil

effects in experimental settings using different routes of administration shows very

promising results for the potential management of a variety of pathological conditions including bacterial and fungal infestation, cardiovalcular conditions and cancer. The

main effect of Bergamot essential oil as a neuroprotective in cancer was explained

9

REFERENCES

Abdul Rahman Bin Abdul Ghafar. 2013. Optimisation of kaffir lime leaves (citrus hystrix) volatile oil extraction by pressurised liquid extraction (pie) using response surface methodology (rsm). Faculty of Applied Sciences. Universiti Teknologi MARA.

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