Study on extraction, chemical composition and antioxidant capacity of cinnamomum burmannii essential oil

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY VU THI THUY STUDY ON EXTRACTION, CHEMICAL COMPOSITION AND ANTIOXIDANT CAPACITY OF CINNAMOMUM BURMANNII ESSENTIAL OIL BACHELOR THESIS Study mode : Full time Major : Food technology Falcuty : International Program Office Batch : 2016 - 2020 THAI NGUYEN, 2022 ABTRACT Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Food Technology Student name Vu Thi Thuy Student ID DTN1654190005 Thesis Title Supervisor(s) Study on extraction, chemical composition and antioxidant capacity of Cinnamomum burmannii essential oil MSc Dinh Thi Kim Hoa MSc Nguyen Thuong Tuan Supervisor’s Signature Abstract: The aims of this study was to develop a suitable technological process for extracting essential oil from Cinnamomum burmannii leaves by steam distillation method In addition, the quality, chemical compositions and antioxidant capacity of the essential oil also were evaluated The most suitable parameters for extraction process were the wilting time of 84 hours; the ratio between material and distill water of 5/50 (w/v); 50 minutes for extraction The amount of obtained essential oil was 0.6 - 0.7% The chemical ingredient of essential oil consisted of 23 compounds that were reported for chemical structures The antioxidant activity of the essential oil from Cinnamomum burmannii leaves was revealed with the IC50 of 41.10 (µg/ml) Keywords Extraction, Chemical composition, Antioxidant activity, Cinnamomum burmannii essential oil Number of pages 64 Date of Submission June 1st, 2022 ii ACKNOWLEDGEMENT During my studies and internship at Thai Nguyên University of agriculture and forestry, I have now completed my graduation thesis In order to complete this thesis, I have been guided by the devoted guidance of my supervisor, along with the help of Thai Nguyen University of Agriculture and Forestry, the Institute of Life Sciences, and the Advanced Program Office I also received enthusiastic cooperation from colleagues, help, and encouragement from my family members In response to that sentiment, though here I would like to express my deep gratitude and respect to all collectives and individuals who created conditions to help me throughout the internship process First of all, I would like to express my sincere thanks to the school management board, the Dean of the Faculty of Food Technology, and the collective of teachers in the Faculty of Food Technology, Thai Nguyen University of Agriculture and Forestry that taught and mentored me during these time, as well as my graduation internship In particular, I sincerely thank for the attention and guidance of the instructing supervisor MSc Dinh Thi Kim Hoa and MSc Nguyen Thuong Tuan, who directly guided me to implement this thesis successfully Through this, I would also like to express my gratitude to family, relatives, and friends for helping and encouraging me during my study and practice at school Finally, I would like to respectfully send my sincere thanks and best wishes to the teachers and teachers on the evaluation committee Thank you sincerely! Thai Nguyen, May 23, 2022 Student VU THI THUY iii TABLE OF CONTENT LIST OF FIGURES LIST OF TABLE ABBREVIATIONS PART I INTRODUCTION 1.1 Research rationale 1.2 Research objective 1.3 Detail goals 1.4 Limitations PART II LITERATURE REVIEW 2.1 Overview of Cinnamomum burmannii tree 2.1.1 Characteristic of Cinnamomum burmannii 2.1.2 Distribution, ecological characteristic and growth 2.1.3 Uses of Cinnamomum burmannii leaves 2.2 Overview of essential oil 10 2.2.1 Definition of essential oil 10 2.2.2 Physical and chemical properties of essential oil 11 2.2.3 The role of essential oils in plant life 12 2.2.4 Biological activities and uses of essential oils 14 2.2.5 Cinnamomum burmannii essential oil 15 2.3 Methods extraction of essential oil 17 2.3.1 Hydrodistillation 18 2.3.2 Solvent extraction 19 2.3.3 Enfleurage 21 2.3.4 Maceration 21 2.3.5 Cold pressing extraction 21 PART III METHODOLOGY 23 3.1 Material and research scope 23 3.1.1 Material 23 3.1.2 Research scope 23 3.2 Work place and time to proceed 23 3.3 Equipment and chemicals 24 3.4 Research content 25 iv 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 3.4.1 The research on factors affecting the content of Cinnamomum burmannii essential oil 25 3.4.2 Determination of the chemical composition of Cinnamomum burmannii essential oil by GC-MS method 25 3.4.3 Investigation of antioxidant capacity of essential oils 26 3.5 Research methods 26 3.5.1 The process of extracting essential oil from Cinnamomum burmannii leaves 26 3.5.2 Experimental design method 27 3.5.3 Determination of the chemical composition of Cinnamomum burmannii essential oil by GC-MS method 29 3.5.4 Investigation of antioxidant capacity of essential oil 29 3.5.5 Determination of moisture and ash content in Cinnamomum burmannii leaves 31 3.5.6 Determination of some physicochemical parameters of Cinnamomum burmannii essential oil 33 3.6 Data satistical analysis methods 36 PART IV RESULT AND DICUSSION 37 4.1 Moisture and ash content in Cinnamomum burmannii leaves 37 4.2 The result of studying factors affecting the distillation process of essential oils 37 4.2.1 Effect of extraction time 37 4.2.2 Effect of the material/distilled water ratio on essential oil content 38 4.2.3 Effect of wilting time of raw material on essential oil content 39 4.4 Result for determining the chemical composition of Cinnamomum burmannii essential oil by GC-MS method 41 4.5 Result for determining the antioxidant capacity of Cinnamomum burmannii essential oil 42 4.6 Result for determining some physicochemical of essential oils 44 4.7 Completing the extraction process of Cinnamomum burmannii essential oils 46 PART V CONCLUSION 48 5.1 Conclusion 48 5.2 Recommendation 48 REFERENCES 50 APPENDIXES 55 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 v 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 LIST OF FIGURES Figure 2.1 Cinnamomum burmannii Tree Figure 2.2 Main molecules of essential oils 10 Figure 3.1 Cinnamomum burmannii leaves 23 Figure 3.2 The process of extracting from Cinnamomum burmannii leaves 26 Figure 3.3 Reaction mechanism of 2,2-diphenyl-1-picrylhydrazyl (DPPH) with antioxidant 30 Figure 4.1 Graph showing the effect of time on essential oil content 38 Figure 4.2 The graph shows the effect of the amount of distilled water on the essential oil content 39 Figure 4.3 Graph showing the effect of wilting time on essential oil content 40 Figure 4.4 Correlation between free radical inhibitory activity and concentration of Cinnamomum burmannii essential oil 43 Figure 4.5 Correlation between free radical inhibitory activity and concentration of Ascorbic acid 43 Figure 4.6 Cinnamomum burmannii essential oil 45 Figure 4.7 The extraction process of Cinnamomum burmannii essential oils 46 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 LIST OF TABLE Table 3.1 Experiment Chemicals 24 Table 3.2 Experiment Equipment 24 Table 3.3 Laboratory instruments 25 Table 3.4 The experimental design to determine the effect of extraction time on essential oils 27 Table 3.5 Experimental design to determine the effect of (material/distilled water) ratio on essential oils 28 Table 3.6 The experimental design to determine the effect of wilting time of raw material on essential oils 28 Table 4.1 The moisture and ash content of Cinnamomum burmannii leaves 37 Table 4.2 Results of survey on extraction time of essential oils 37 Table 4.3 Result of effecting the material/distilled water ratio on essential oil content 38 Table 4.4 Results affect the wilting time of raw material on essential oil content 40 Table 4.5 Chemical composition of Cinnamomum burmannii essential oil 41 Table 4.7 Antioxidant activity of Ascorbic acid with DPPH 43 Table 4.8 Some physicochemical of Cinnamomum burmannii essential oils 45 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37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 ABBREVIATIONS ABTS 2,2'-azino-bis (3-ethylbenzothiazoline-6sulfonic acid) DPPH 2,2-diphenyl-1-picrylhydrazy GC-MS Gas Chromatography-Mass Spectrometry HPLC High-Performance Liquid Chromatography IA Acid index IC50 Half Maximal Inhibitory Concentration IE Saponification index IS Ester index SD Standard Deviation SDE Simultaneous Distillation Extraction SFE Supercritical Fluid Extraction SPME Solid-phase Micro-extraction W/V Weight/Volume 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37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 PART I INTRODUCTION 1.1 Research rationale Essential oils are natural ingredients Currently, in the world, people are increasingly interested in medical products derived from essential oils to ensure health and have no negative effects As a result, both domestic and international research on essential oils is rising in number and quality Vietnam is one of the nations in the tropical monsoon area that has a large and diversified collection of essential oils and aromatic plants, with over 300 essential oil species collected such as Peppermint (1.3%-2%), Gratissimum (0.9%-1.67%), Lemongrass (0.7%-1.5%), Rosemary (0.3%-0.5%), Cinnamon (1%-2.5%), They are used quite commonly in the market due to they mostly contain the main components such as cinnamic aldehyde, geraniol, menthol, borneol, citral, citronellal, menthol, … that pleasant aroma, anti-cold effect, antiinflammatoryand stress reduction Some essential oils can cure cancer such as Frankincense, Lemongrass, and Orange Cinnamomum burmanniiis a woody plant in Bao Lac district, Cao Bang province Cinnamomum burmanniileaves are often used by people as raw materials for food processing In addition, people often boil the leaves for a bath or sauna to prevent and treat diseases such as malaria, and bacterial infections, to control blood pressure and headaches Based on the above applications, we can see that Cinnamomum burmannii leaves are a useful ingredient for health, not only as a food additive but also as a common healing method for people However, these effects are insufficient to explain the phenomenon clearly from a scientific standpoint, and up to this point, there have been no in-depth studies on this leaf Therefore, I conducted the topic: “ Study on extraction, chemical composition and antioxidant capacity of Cinnamomum burmannii essential oil.” Thereby contributing to creating more directions and to clarify for the application of Cinnamomum burmanniiessential oil in the fields of medicine, pharmaceuticals, and cosmetics Simultaneously, maximizing the potential and improving the value of Cinnamomum burmannii essential oil on the market 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37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 1.2 Research objective Develop extraction procedure, investigate the chemical composition, and test the biological activity of the obtained Cinnamomum burmannii essential oil 1.3 Detail goals - Conduct an experiment to extract Cinnamomum burmannii essential oil by direct steam distillation method - Determination of some physicochemical indexes and chemical compositions of essential oils - Dertermination of antioxidant properties of Cinnamomum burmannii essential oil 1.4 Limitations Limitations that are expected to be encountered throughout the study: - Raw materials: Because Cinnamomum burmannii leaves are available in Bao Lac town, Cao Bang province, it is difficult to collect and transport raw materials - There is currently no study on the plant's composition and activities of Cinnamomum burmannii 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 REFERENCES English document Al-Dhubiab, B E (2012) Pharmaceutical applications and phytochemical profile of Cinnamomum Burmannii Pharmacognosy reviews, 6(12), 125 Abid Shah SF and Bashir S (2019) Comparative Proximate Analysis of Cinnamon And Lemongrass for its Nutritive Value JSM Human Nutri Food Sci 5: 3 Atti-Santos, A C., Rossato, M., Serafini, L A., Cassel, E., & Moyna, P (2005) Extraction of essential oils from lime (Citrus latifolia Tanaka) by hydrodistillation and supercritical carbon dioxide 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37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 Streptococcus pyogenes and Pseudomonas aeruginosa Int J Curr Microbiol App Sci, 3(2), 177-178 11 Cheng B, Xu Y, Zeng F, Yu X, Ding J, Wu Y, et al Chemical constituents of essential oil of Cinnamomum burmanniif heyneanum Yunnan Zhiwu Yanjiu 1992;14:105–10 12 Christenhusz, M J M.; Byng, J W (2016) "The number of known plants species in the world and its annual increase" Phytotaxa 261 (3): 201–217 13 Deng CC, Huo LN, Li PY, et al Chemical constituents and antioxidant activity of essential oils from 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37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 22 Kuspradini, Harlinda, et al "Bioactivity of essential oils from leaves of Dryobalanops lanceolata, Cinnamomum Burmannii, Cananga odorata, and Scorodocarpus borneensis." 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Tạp chí Nghiên cứu khoa học Phát triển kinh tế Trường Đại học Tây Đô 06: 190-201 47 Nguyễn Thị Thu Thảo, Phùng Thị Lan Hương, Hoàng Thị Lý (2021) Nghiên cứu thành phần hóa học ứng dụng sản xuất dầu cao xoa tinh dầu quế trồng tỉnh Phú Thọ Study Aids & Test Prep Documents 48 Phát, Đ T., Quyên, N T C., Ngân, T T K., Nhựt, P T., Hiền, T T., Việt, N T., & Cường, L X (2019) Nghiên cứu mô hình động học trình chiết xuất thành phần hóa học tinh dầu vỏ Cam (Citrus sinensis) Journal of Science and Technology, 2(4), 19-25 49 Phúc, N Đ., Danh, P H., Anh, T T., & Phương, T H Y (2020) Chiết xuất tinh dầu Hương thảo phương pháp chưng cất nước Journal of Science and Technology, 3(2), 21-25 Internet document 50 https://duoclieuvietnam.org/tinh-dau-thuc-vat-va-nhung-ung-dung-trong-doisong-trang-thong-tin-duoc-lieu-cay-thuoc-vi-thuoc-viet-nam/ 51 The Seasoning and Spice Association, 2010 Culinary Herbs and Spices Available from:http://www.seasoningandspice.org.uk/ssa/background_culinary-herbsspices.aspx 52 https://caythuoc.org/que-ranh-tren-tren-dieu-tri-phong-thap-dau-nhucxuong-khop-va-chan-an.html 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 54 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Mean Sample Size = 3.000 Table A.2: Tables of data processed by SPSS software of the effect of distilled water volume on essential oil content ANOVA Essentialoilcontent Sum of Squares df Mean Square Between Groups 049 012 Within Groups 000 10 000 Total 049 14 F 1219847.233 Sig .000 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 55 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 Essentialoilcontent Duncan Volumeofdistillwater N Subset for alpha = 0.05 1.0000 2.0000 5.0000 3.0000 4.0000 3 439301 520067 549053 560737 612702 Sig 1.000 1.000 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 Table A.3: Tables of data processed by SPSS software of effect of sample wilting time on essential oil content ANOVA Essentialoilcontent Sum of Squares df Mean Square Between Groups 069 017 Within Groups 000 10 000 Total 069 14 F Sig 2865178.722 000 Essentialoilcontent Duncan wiltingtime N Subset for alpha = 0.05 5.0000 1.0000 2.0000 3.0000 4.0000 Sig 537534 577983 606900 682010 722533 1.000 1.000 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 56 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.2237.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.66 Table A.4: Tables of data processed by SPSS software of Antioxidant activity of Cinnamomum burmanniiwith DPPH ANOVA Dpph Sum of Squares Between Groups Mean Square 1035.975 588 10 059 4144.487 14 Within Groups Total df 4143.900 F Sig 17630.615 000 Dpph Duncan factor N Subset for alpha = 0.05 1.00 2.00 3.00 4.00 5.00 3 32.5521 45.6011 63.9890 76.4201 89.2322 Sig 1.000 1.000 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 Table A.5: Tables of data processed by SPSS software of antioxidant activity of Ascorbic acid with DPPH ANOVA Ascorbic acid Sum of Squares Between Groups Mean Square 4011.432 1002.858 007 10 001 4011.439 14 Within Groups Total df F 1519481.944 Sig .000 Ascorbic acid Duncan factor N Subset for alpha = 0.05 1.00 2.00 3.00 4.00 5.00 Sig 39.8153 49.0517 59.0333 74.8700 84.4801 1.000 1.000 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 57 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37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 58 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37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.99 59 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