VIETNAM NATIONAL UNIVERSITY, HA NOI VNU UNIVERSITY OF SCIENCE FACULTY OF CHEMISTRY NGUYEN THI THU NGOC EUGENOL – CHEMICAL COMPOSITION OF CLOVES AND SOME DERIVATIVES WITH HIGH USE Instructor Dr Nguyen Hoang Yen Ha Noi – 2021 2 ACKNOWLEDGEMENT First of all, I would like to express my gratitude and sincere thanks to Dr Nguyen Hoang Yen has enthusiastically helped and oriented his way of thinking and working scientifically Those are very valuable suggestions not only in the process of making thi.
VIETNAM NATIONAL UNIVERSITY, HA NOI VNU UNIVERSITY OF SCIENCE FACULTY OF CHEMISTRY *** NGUYEN THI THU NGOC EUGENOL – CHEMICAL COMPOSITION OF CLOVES AND SOME DERIVATIVES WITH HIGH USE Instructor: Dr Nguyen Hoang Yen Ha Noi – 2021 ACKNOWLEDGEMENT First of all, I would like to express my gratitude and sincere thanks to Dr Nguyen Hoang Yen has enthusiastically helped and oriented his way of thinking and working scientifically Those are very valuable suggestions not only in the process of making this thesis, but also a stepping stone for me in the process of studying and setting up a career later Next, I would like to thank my family and friends, who are always willing to share and help in study and life Hope we will stick together forever Wish the best will always accompany everyone Thank you for all Student, Nguyen Thi Thu Ngoc CONTENT ACKNOWLEDGEMENT CONTENT SIGNAL TABLE OF ABBREVIATIONS INTRODUCTION CHAPTER 1: OVERVIEW Overview of clove 1.1 Scientific name 1.2 Description of clove 1.3 Distribution 1.4 Plant and harvest 1.5 Parts used to separate essential oils Effect Chemical composition of clove Eugenol 10 CHAPTER 2: EUGENOL EXTRACT FROM CLOVE OIL 11 CHAPTER 3: SOME DERIVATIVES OF EUGENOL 13 Isoeugenol 13 Zinc oxyde eugenol 14 Some derivatives of eugenol 15 CHAPTER 4: FUTURE RESEARCH DIRECTIONS 16 REFERENCES 17 SIGNAL TABLE OF ABBREVIATIONS Abbreviation Meaning DPPH 2,2-Diphenyl-1-picrylhydrazyl ABTS 2,2’-Azino-bis(3-ethylbenzothiazoline-sulfonic acid) DMBA 9,10-Dimethylbenz-A-Anthracene CH2Cl2 Dichloromethan ZOE Zinc oxyde - Eugenol INTRODUCTION Essential oils are an extremely valuable gift that nature bestows on humans Each essential oil contains tens to hundreds of different natural compounds, so essential oils are being considered as an excellent choice in therapy, aromatherapy and beauty One of the essential oils that is in great demand today is Syzygium aromaticum (clove) oil The main component of clove is eugenol (2-methoxy-4-allylphenol) It makes up about 76.8% of the composition of clove essential oil and has a very pleasant smell Eugenol has many uses: Eugenol in dental medicine is used as a toothache, with oxide e zinc Eugenol cement forming adhesive substance used for temporary fillings In the food industry, eugenol is used as a raw material for the synthesis of vanillin From Eugenol synthesized methyl e ugenol, a substance that attracts insects to gather in one place to easily trap them , and synthesize a number of substances that stimulate plant growth In addition, eugenol also plays an important role in the cosmetic industry In this yearbook, we want to clarify three things: - Extract of Eugenol from Clove - Synthesis of some derivatives of eugenol - Preliminary exploration of the biological effects of eugenol and other substances CHAPTER 1: OVERVIEW Overview of cloves 1.1 Scientific name: Syzygium aromaticum (L.) Merrill &L M Perry Tên đồng nghĩa: Eugenia caryophyllata Thunb Cloves are also known as Cong cloves, Hung Tu Huong, and Dinh Tu Scientific classification: Gender: Plantae Set: Myrtales Family: Myrtaceae Genus: Eugenia Species: E.caryophyllata Thunb 1.2 Description of clove: Figure Cloves + Small tree, 10-12m tall Leaves opposite, oval , pointed, not falling Figure Clove flower + Flowers grow into small cymbal and branched at the top of branches, flowers consist of thick sepals, bright red when ripe, pink petals, falling early when blooming, lots of stamens + Fruits are long berries, surrounded by sepals, usually containing only one seed 1.3 Distribution: Cloves are native to the Moluccas Islands (Indonesia) and are produced mainly in Tanzania, Malysia, Indonesia, the Zanziba Islands, Pemba (Tandania), Madagascar Our country used to import cloves Recently, we have tried to migrate a few trees but have not developed and are currently losing seed first.1 1.4 Plant and harvest: Planted with seeds The 5th or 6th year will flower, but the highest harvest is in the 20th year Depending on the region, each year will be harvested 1-2 times, when the buds begin to red Pick flowers by hand while still in the bud stage, separating the stem and flower buds Hang or dry until it turns brown Do not wait until the fruit is harvested before harvesting because then the quality is much worse when in bud form 1.5 Parts used to separate essential oils Oleum Caryophyllorum commercial name is clove oil, is distilled from flower buds, flower stalks and leaves 90-95% of clove leaf essential oil is distilled from the leaves, usually only poor quality flower buds are used to store the essential oil Effect Since ancient times, the emperors of the Han Dynasty used to make their followers chew clove leaves to freshen their breath Found since ancient times, has become a valuable commodity and has caused a fierce competition between the Portuguese, Dutch and British for the position of the spice trading market Because Clove flower has been considered a precious spice to process curry powder along with turmeric, ginger, black pepper, chili Clove flowers are also used to marinate aromatic tobacco and make liqueurs, perfume Today, in many Southeast Asian countries, people still have the custom of using clove flowers for the purpose of freshening their breath, fighting bad breath and preventing diseases during outbreaks In our country, Hai Thuong Lan Ong also used clove flowers in combination with cinnamon, raw ginger, bone pain, bone anise, seashells, water shells, chau chau leaves, cactus latex, purple mulberry leaves jasmine leaves, vajra leaves, buying leaves, blood senses, turmeric, chyme seeds, dog blood seeds, pomelo leaves, star fruit mistletoe leaves, equal amounts of 10-20g, pound hot stars and apply to cure sprains , luxation Eugenol is the main component of the essential oil extracted from Clove flower buds It was the first component of an essential oil that has been shown to act as an antiseptic and pain reliever in dentistry that is still used today Eugenol is also an important agent in cosmetics and food flavoring Its properties towards various radicals such as DPPH, ABTS, superoxide and azide, hydroxyl and aloperoxyl radicals have been reported It has been shown to inhibit prostaglandin synthesis-7 and COX-2 inhibition value IC50 is 129 mol ( The IC50 value of was used to compare the potentials of the two antagonists) In carcinogenicity testing of different groups in CD-1 and F344 mice Eugenol did not bind to the tumor In the "skin paining" study by Van Duuren and Gold-schmidt, Eugenol effectively contributed to the inhibition of benzopyrene, a skin carcinogen Eugenol can inhibit DMBA-generine oil-causing cancer by about 84% (DMBA is a chemical commonly used to induce experimental breast cancer.) In the Bl6 Eugenol therapy resulted in a significant reduction in tumor growth (p = 0.0057), a nearly 40% reduction in tumor size, and a 19% increase in mean time to endpoint Many scientific evidences have proven that Eugenol has cytotoxic potential, which can be used to treat human cancer cells Chemical composition of cloves Clove essential oil with content of 15-20% in flower buds, 5-6 % in flower stalks and 2-3 % in leaves.1 The composition of clove essential oil is listed in the following table: Compound Percent limonene 0.1 1,8-cineole 0.1 methyl salicylate 0.1 methylchavicol 0.2 chavicol 0.1 eugenol 76.8 -caryophyllene 17.4 trans-isoeugenol 0.1 R-humulene 2.1 R-farnesene 0.1 eugenyl acetate 1.2 caryophyllene alcohol 0.1 caryophyllene oxide 0.4 R-humulene epoxide 0.1 Table 1.Chemical composition of clove essential oil Density: d15 = 1.043 – 1.068 Refractive Index: D20 = 1.529 – 1.537 Maximum productivity rotary: D20 = -1o35 From the table above we can see , the main component of clove essential oil is eugenol (76.8%), followed by −caryophyllene (17.4%), R-humulene (2.1%), eugenyl acetate (1.2%).9 Eugenol: • Chemical formula: Common name: eugenol Nomenclature name: 4- allyl-2methoxy-phenol hay 2- methoxy-4 (2 propenyl) - phenol • Physical properties: Colorless or pale yellow, liquid, transparent, darkened by exposure to air Has a characteristic smell of cloves Solubility: Practically insoluble in water, soluble in 70% ethanol, insoluble in glycerol, miscible with ethanol 96%, with glacial acetic acid, with methylene chloride and with fatty oils Molecular Mass: 164.20 g/mol Density: 1.06 g/cm3 Melting temperature: -9oC Boiling temperature: 256oC Refractive Index: 1.540 – 1.542 10 • Chemical properties: Eugenol is an aromatic ring substance with a phenol functional group , containing a double bond and an ether Evaluate the characteristic reaction of phenol: with FeCl3, with NaOH to form a phenolate salt Take part in the characteristic reaction of an ether Join the addition reaction 10 CHAPTER 2: EUGENOL EXTRACT FROM CLOVE ESSENTIAL OIL - Use organic solvents Figure 3: Steam entrainment distillation unit Assemble a steam entrainment distiller as figure Put about 5g of Clove buds in a 100ml flask Add ml of water and some boiling ice Mark the water level to replenish the water lost during distillation and prevent the sample from drying out Heat the mixture until the distillation begins to stabilize Check the water level and add water as needed When about 20 ml of distillate has been collected, stop heating and transfer the product to a 125 ml separatory funnel Rinse the flask with ml of CH2Cl2 and add to the separating funnel Extract Eugenol with CH2Cl2 and collect the extract in a 25 ml erlenic flask Repeat the extraction with 5ml CH2Cl2 each time The dried extracts with Na2SO4 and smoke extract made anhydrous by pipette Evaporate the solvent to obtain pure eugenol.11 - Or extracting eugenol is based on the principle : Eugenol with OH of phenol when reacting with alkali will form alkaline eugenate This salt is soluble in water, insoluble in other components of the essential oil Based on this property, it is possible to separate alkaline eugenate from the unreacted essential oil Convert alkaline eugenate to eugenol by acid Purified by steam distillation or fractional distillation at reduced pressure ▪ Shake the essential oil with KOH Separate the potassium eugenate fraction Convert eugenate to eugenol with HCl solution Wash crude Eugenol with CaCO3 Purify by steam distillation.3 ▪ Use 13-14% NaOH to convert eugenol to eugenate, separate the sodium eugenate Use HC1 to convert eugenate to eugenol Distill the fraction at reduced pressure of 18-20 mmHg, take the fraction with boiling point at138 - 140 °C 11 ▪ Use KOH to convert eugenol to eugenate The eugenate separation was carried out at 80 - 90°C Acidification with H2SO4 to convert eugenate to Eugenol Fractional distillation at 139°C yields pure eugenol 12 CHAPTER 3: SOME DERIVATIVES OF EUGENOL Isoeugenol Isoeugenol is rare in the composition of natural essential oils It has a milder fragrance than eugenol and is included in several fragrances Isoeugenol has a stronger antifungal effect than Eugenol.6 Isoeugenol is the starting material for the synthesis of several aromatic substances, typically vanillin Isoeugenol is usually prepared by isomerization of eugenol in an alkaline medium at high temperature 11 Eugenol Isoeugenol To explain the possible difference in biological activity and reactivity of Isoeugenol compared with that of eugenol, it is suggested that the transfer of the double bond closer to the aromatic nucleus in the Isoeugenol molecule forms a conjugate chain, thereby affecting electron system in the benzene ring Nowadays, by quantum chemical method, it is possible to calculate the charge distribution of atoms in the molecule, showing that the conversion of the allyl group to propenyl has clearly changed the charge distribution at positions 3, and in the benzene ring The commercial Isoeugenol product prepared from Eugenol consisting of a mixture of cis and trans forms has attracted much attention because the trans-Isoeugenol product has a milder aroma than cis-Isoeugenol and is used as a better flavoring agent Trans isoeugenol Cis isoeugenol Characteristics: - Isoeugenol oily liquid is pale yellow, slightly fragrant than Eugenol 5, odor similar to carnations - Density: 1,079 – 1,085 (g/cm3) - Extract: 1,5720 – 1,5770 - Phenolic content: 99,5% - Boiling point: 266oC 13 Zinc oxyde – Eugenol (ZOE): For many years, the dental industry has used the complex between ZnO and Eugenol as a temporary filling material 12 ZnO can t Create complex with substances tamarind group t hoxy in ortho position compared with phenol OH: Eugenol Guaiacol Methylguaiacol With Eugenol, ZnO complexed according to the following equation: In dentistry, ZOE is used in the simple way that ZnO powder is mixed with Eugenol in a 1:1 ratio to form a homogeneous fine powder This fine powder used for temporary fillings is called Zinc Eugenat It is often used in dentistry when the decay is very deep or very close to the nerve or pulp chamber For persons with a dry socket as a complication of tooth extraction, packing the dry socket with a eugenol-zinc oxide paste on iodoform gauze is effective for reducing acute pain.13 The placement of a ZOE "temporary" for a few to several days prior to the placement of the final filling can help to sedate the pulp But, ZOE had in vitro cytotoxicity majorly due to release of Zn ions.14 In spite of severe in vitro cytotoxicity, ZOE showed relatively good biocompatiblity in animal study when ZOE was applied on dentin.15 When ZOE was used as dentin-protective based materials, use of dental composite resin on ZOE was strongly prevented due to its inhibition of resin polymerization through radical scavenging effect.16 It is classified as an intermediate restorative material and has anaesthetic and antibacterial properties The exact mechanism of anesthetic effect from ZOE was not revealed perfectly, but possibly through anti-inflammatory effect, modulating immune-cells to less inflamed status.17 14 Some eugenol derivatives have special properties ` Dihydroeugenol Derivatives Function Substitute nitrogen at position of dihydroeugenol Sodium salt of dihydroeugenol Pain relief, no skin irritation 18 Slight drop in blood pressure 18 Dihydroeugenol derivative with an amine group attached to the position Cure bleeding but quite toxic 19 Esters of eugenol with carboxylic acids Significant pain relief 25 Mixture of eugenol and isoeugenol Cure AIDE 20 Axetyleugenol Axetyisoeugenol Cosmetic flavoring , with the smell of almonds 21 Eugenol acetate Antibacterial and anti-virulence against drug-resistant Acinetobacter baumannii 22 Methyleugenol Methyl isoeugenol Anti-fungal, capable of repelling many types of insects 23 Vanillin Flavoring agents in foods, beverages and pharmaceuticals 24 Table 2: Some eugenol derivatives have special properties 15 CHAPTER 4: FUTURE RESEARCH DIRECTIONS Eugenol is a naturally occurring phenolic molecule found in some plants It has antiinflammatory, antioxidant, and pain-relieving properties Eugenol has many uses - Relieve toothache, mouth ulcers: - Kills Candida fungus - Used in the production of stabilizers and antioxidants for plastics and rubber - Strong antibacterial - Reduces the presence of Listeria monocytogenes and Lactobacillus sakei bacteria in food As an anesthetic: development of eugenol derivatives as intravenous anesthetics, as an alternative to propanidide We can see that eugenol is extracted to serve many different industries for life We see that eugenol's properties are anti-inflammatory and antioxidant, so it can be applied in cosmetics , creating cleansers, shower gels, and exfoliating scrubs in people with acneprone skin Eugenol can destroy plankton cells, and biofilms of Staphylococcus Aureus bacteria that cause acne, boils, skin sores, rashes, blisters, impetigo and Staphylococcal burn skin syndrome The eugenol in clove oil has rejuvenating and stimulating properties, which can increase blood flow to the skin and make it look radiant reducing the effects of aging, like wrinkles, sagging skin, and blemishes Eugenol has antiseptic properties, so it is very useful for people who often pop pimples Not only that, it has a characteristic scent that is quite fragrant, helping people to feel comfortable From the above uses, I would like to study to clarify the antibacterial properties of eugenol and its derivatives : Extraction of eugenol from clove essential oil Preparation of eugenol and their derivatives Testing the antibacterial activity of eugenol and its derivatives 16 REFERENCES - References Vietnamese Bộ Y tế, Bộ Giáo dục Đào tạo (1998),Bài giảng dược liệu tập II, Ha Noi Hải Thượng Lãn Ông, Thuốc Việt Nam, Nhà xuất Văn hóa dân tộc Phạm Trương Thị Thọ (1996), “Điều kiện tối ưu chiết xuất eugenol từ tinh dầu hương nhu” ,Tạp chí khoa học cơng nghệ , (2), pp 28 -33 Giang Thị Sơn (1996) , “Điều chế eugenol có độ tinh khiết cao làm nguyên liệu bán tổng hợp dãn chất eugenol”, Tạp chí dược học, (5), pp 8-9 Nguyễn Thị Hoàng Oanh, Tách eugenol từ tinh dầu hương nhu trắng chuyển hoá thành số sản phẩm hương liệu ,Graduation essay , Vietnam National University, Hanoi Phạm Trương Thị Thọ (1994), Nghiên cứu bán tổng hợp định hướng dẫn xuất eugenol có tác dụng kháng nấm Candida albican , Science doctoral thesis, Hanoi - References English Yanling Geng, Jianhua Liu, Ruimin Lv, Jinpeng Yuan, Yunliang Lin and Xiao Wang (2007), “An efficient method for extraction, sepration and purification of Eugenol from Eugenia caryophyllata by supercritical fluid extraction and high-speed counter-current chromatography” Chae-Bin Yoo, Ki-Tae Lee, Kyu-Seok Cho, Joohun Ha, Hee-Jun Park, JungHwan Nam, Uk-Hyun Kil and Kyung-Tae Lee (2005), “Eugenol isolated from the essential oil of Eugenia caryophyllata induces a reactive oxygen speciesmediated apoptosis in HL-60 human promyelocytic leukemia cells” Leopold Jirovetz, Gerhard Buchbauer, Ivanka Stoilova, Albena Stoyanova, Albert Krastanov and Erich Schmidt (2006), “Chemical Composition and Antioxidant Properties of Clove Leaf Essential Oil”, Journal of Agricultural and Food Chemistry, 54, 17, pp 6303-6307 10 British pharmacopoeia (2009) 11 Cerritos Community College, Extraction and purification of Eugenol 12 Kick Othmer , Encyclopedia of Chemical Technology, 6, pp 779, 823, 824 13 Tarakji B, Saleh LA, Umair A, Azzeghaiby SN and Hanouneh S ( 2015), "Systemic review of dry socket: aetiology, treatment, and prevention" J Clin Diagn Res, (4) 14 Lee Jung-Hwan; Lee Hae-Hyoung; Kim Kyoung-Nam and Kim KwangMahn (2016), "Cytotoxicity and anti-inflammatory effects of zinc ions and eugenol during setting of ZOE in immortalized human oral keratinocytes grown as three-dimensional spheroids", Dental Materials, 32 (5), pp.93 – 104 15 Sabir Ardo; Tabbu Charles R; Agustiono Purwanto and Sosroseno Wihaskoro (2005), "Histological analysis of rat dental pulp tissue capped with propolis" Journal of Oral Science, 47 (3), pp 135–138 16 Carvalho Ceci Nunes; Bauer José roberto De Oliveira; Loguercio Alessandro Dourado and Reis Alessandra (2007), "Effect of Zoe Temporary Restoration on Resin-Dentin Bond Strength Using Different Adhesive Strategies", Journal 17 of Esthetic and Restorative Dentistry, 19 (3), pp 144–152 17 Lee Jung-Hwan; Lee Hae-Hyoung; Kim Hae-Won; Yu Je-Wook; Kim Kyoung-Nam; Kim Kwang-Mahn (2017), "Immunomodulatory, antiinflammatory effect of ZOE-based dental materials", Dental Materials, 33 (1), pp 1–12 18 Brady Dann ( 1915), Nitrovanilin – J Chem Soc , 107, 1861 19 David E Levin and Alexander Lowy, (1993), “Derivatives of Dihydroeugenol and Certain Pharm properties of Some of the compounds”, J Am Chem Soc, 55, pp.1995-2000 20 Kaempgen dicter (1990), Eugenol-isoeugenol mixtures for Treatment of AIDS Ger.offen 21 Steffen Arctander (1969), Perfume and Favor Chemical Montclair N.J, USA, pp 198 22 Syed MK & Voravuthikunchai SP (2016), Flavour and Fragrance Journal 31(6), pp.448-454 23 Tan Keng Hong and Nishida Ritsuo (2012), "Methyl Eugenol: Its Occurrence, Distribution, and Role in Nature, Especially in Relation to Insect Behavior and Pollination", Journal of Insect Science, 12 (56), pp 1–60 24 Fund for Research into Industrial Development, Growth and Equity (FRIDGE) (2004), Study into the Establishment of an Aroma and Fragrance Fine Chemicals Value Chain in South Africa, Part Three: Aroma Chemicals Derived from Petrochemical Feedstocks, National Economic Development and Labor Council - French references 25 Mathieu Andre & Blum Jean (1979), Demande de Brevet D’invention No79 24051, Paris 18 ... độ tinh khiết cao làm nguyên liệu bán tổng hợp dãn chất eugenol? ??, Tạp chí dược học, (5), pp 8-9 Nguyễn Thị Hoàng Oanh, Tách eugenol từ tinh dầu hương nhu trắng chuyển hoá thành số sản phẩm hương... Văn hóa dân tộc Phạm Trương Thị Thọ (1996), “Điều kiện tối ưu chiết xuất eugenol từ tinh dầu hương nhu” ,Tạp chí khoa học công nghệ , (2), pp 28 -33 Giang Thị Sơn (1996) , “Điều chế eugenol có. .. clove Eugenol 10 CHAPTER 2: EUGENOL EXTRACT FROM CLOVE OIL 11 CHAPTER 3: SOME DERIVATIVES OF EUGENOL 13 Isoeugenol 13 Zinc oxyde eugenol