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Kaffir Lime) Leaves Oil

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Kaffir Lime) Leaves Oil 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 BORANGPENGESAHAN STATUS TESIS JUDUL: ACTIVATINGEFFECT OF AROMATHERAPYINHALATION WITH Citrus hystrix (KAFFIR LIME) LEAVESOIL 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 (/): üýtºi6r: iýi:: f 11 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 TERHADyang telah ditentukan oleh organisasi/badan di mana penyelidikan dijalankan x TIDAK TERHAD DisahkanOleh, NURULAINBINTI ISMAIL PUSTAKAWAIWANAN ABAH ALISA STEPHANIERASION (TandatanganPustakawan) MS1521001T Tarikh :V October 2018 (Prof. Madya. Dr. How Siew Eng) Penyelia DECLARATION I declarethat this thesis and the work presentedin it are my own and has been generatedby me as the result of my own original research. 27t'' September2018 Alisa Sianie Rasion MS1521001T 11 CERTIFICATION NAME ALISA STEPHANIE RASION MATRICNO. MS1521001T TITLE ACTIVATING EFFECT OF AROMATHERAPY INHALATION WITH Citrus hystrix (KAFFIR LIME) LEAVES OIL DEGREE : MASTER OF SCIENCE (INDUSTRIAL CHEMISTRY) VIVA DATE 9THAUGUST 2018 CERTIFIED BY; SIGNATURE SUPERVISOR PROF. MADYA.DR. HOW SIEW ENG "nýýö ACKNOWLEDGEMENTS First and foremost, praisesand thanks to the God,the Almighty, for His showersof blessingsthroughout my research work to complete the research successfully.I would liketo expressmy sinceregratitude to my advisorProf. Madya.Dr. How Siew Eng for the continuoussupport of my master study and research,for her patience, motivation,and immense knowledge. Her guidance helped me in all the time of researchand writing of this thesis and finally I am extremelygrateful to my parents and to my friends for providing me with unfailing support and continuous encouragementthroughout my years of study and through the process of researchingand writing this thesis. This accomplishmentwould not have been possiblewithout them. Thank you. Alisa Stephanie Rasion 27thSeptember 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 hystrix that C. 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 data where the 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 has been oil 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 melegakankemurungan dan menaikkanmood manusiadan ini mungkin disebabkan o%h kandungancitronellal yang tinggi di dalam daun minyak pati C. hystrix dan is selamat digunakan. vii TABLE OF CONTENTS Page TITLE i DECLARATION 11 CERTIFICATION III ACKNOWLEDGEMENTS iv ABSTRACT v ASSRRAK VI TABLE OF CONTENTS viii LIST OF TABLES X LIST OF FIGURES XII LIST OF ABBREVIATIONS/ SYMBOLS XIII LIST OF APPENDICES xv CHAPTER 1: INTRODUCTION 1 1.1 Introduction 1 1.2 Significanceof the Study 4 1.3 Scopeof the Study 5 1.4 ResearchObjectives 5 CHAPTER 2: LITERATURE REVIEWS 6 2.1 Aromatherapy 6 2.2 EssentialOil 8 2.3 Compositionof EssentialOil 11 2.4 Actions of Essential Oil On the Body 12 2.5 Aroma-therapeuticEffect of EssentialOil on HumanBehavioural and 14 EmotionalChanges 2.6 OlfactionPhysiology 15 2.7 PsychologicalEvaluation of FragranceEffects Using a Visual Analog 17 Scale(VAS) 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 Comparisonof Controland Treatment Relatedto Six Variables 55 4.3.2 ComparisonBeginning and Ending 56 4.3.3 RelationshipAmong Variables 59 CHAPTER 5: CONCLUSION 63 5.1 Conclusion 63 5.2 Future Research 64 REFERENCES 65 APPENDICES 81 ix LIST OF TABLES Page Table 2.2 Percentagecomposition of leaves 22 oils of Kaffirlime Table3.1 GC-MSconditions for extractedoil 39 analysis Table3.2 Definitionof relaxation,vigorous, 43 calmness,attentiveness, mood and alertness Table4.1 Chemicalcomposition of C.hystrix 45 leavesoil usingGC-MS Table 4.2 Comparisonof chemical 46 compositionof Citrus hystrix leaves oil betweenthe present data and previousdata of the existing literature (%) Table4.3 In vitro cytotoxicityof essentialoil 54 of C. hystrix(leaves) Table4.4 Comparisonof mean rank and sum 55 of rank betweencontrol and C. hystrix leavesoil categoryfor the effects of relaxation,vigorous, calmness,attentiveness, mood and alertness X Table4.5 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 Ratingmean (±standard deviation) 57 and comparisonof rating at the beginningand ending observation time of six variablesrated by 20 respondentsfor the effects of Citrus hytrix leavesoil Table 4.7 Spearman's Rho Correlation 60 between six variables for the effects of Citrus hytrix leaves oil XI LIST OF FIGURES Page Figure2.1 The flow of air inhalation 13 into the nostrils Figure 2.2 Structure of the 27 Citronellal Figure 2.3 Structure of the 29 Citronellol,Citronellal and Hydroxycitronellal 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 Figure4.2 The chromatogramof 50 standard citronella) Figure4.3 The chromatogramof C. 51 hystrix oil confirmedwith
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