EUGENOL THÀNH PHẦN HÓA HỌC CỦA ĐÁNH RĂNG VÀ MỘT SỐ CHẤT KHỬ MÙI CÓ CÔNG DỤNG CAO

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

2

CONTENT

ACKNOWLEDGEMENT............................................................................................. 2
CONTENT...................................................................................................................... 3
SIGNAL TABLE OF ABBREVIATIONS .................................................................. 4
INTRODUCTION ......................................................................................................... 5
CHAPTER 1: OVERVIEW .......................................................................................... 6
1. Overview of clove .................................................................................................... 6
1.1. Scientific name ................................................................................................... 6
1.2. Description of clove ............................................................................................ 6
1.3. Distribution ......................................................................................................... 7
1.4. Plant and harvest .............................................................................................. 7
1.5. Parts used to separate essential oils .................................................................... 7
2. Effect ........................................................................................................................ 7
3. Chemical composition of clove ........................................................................... 8
4. Eugenol .................................................................................................................. 10
CHAPTER 2: EUGENOL EXTRACT FROM CLOVE OIL ................................. 11
CHAPTER 3: SOME DERIVATIVES OF EUGENOL .......................................... 13
1. Isoeugenol............................................................................................................... 13
2. Zinc oxyde eugenol ................................................................................................ 14
3 . Some derivatives of eugenol .............................................................................. 15
CHAPTER 4: FUTURE RESEARCH DIRECTIONS ............................................ 16
REFERENCES ............................................................................................................ 17

3


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

4


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 .

5


CHAPTER 1: OVERVIEW
1. 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 1. Cloves

+ Small tree, 10-12m tall. Leaves opposite, oval , pointed, not falling.

Figure 2. Clove flower
6


+ Flowers grow into small cymbal and branched at the top of branches,
flowers consist of 4 thick sepals, bright red when ripe, 4 pink petals,
falling early when blooming, lots of stamens.
+ Fruits are long berries, surrounded by sepals, usually containing only
one seed. 1

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.

2. 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,
7


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. 2
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. 7 ( 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. 8

3. 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 9 is listed in the following table:

8


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. 1
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

9


4. 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..... 1

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 3.
Put about 5g of Clove buds in a 100ml flask. Add 5 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 5 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. 4
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. 5

12


CHAPTER 3: SOME DERIVATIVES OF EUGENOL

1. 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. 6
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, 5 and 6 in the benzene ring. 6
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. 5

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


2. 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


3. Some eugenol derivatives have special properties.

4.

`

Dihydroeugenol

Derivatives

Function

Substitute nitrogen at position 5 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 5 . 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 :
1.
Extraction of eugenol from clove essential oil
2.
Preparation of eugenol and their derivatives
3.
Testing the antibacterial activity of eugenol and its derivatives

16


REFERENCES
-

References Vietnamese
1. Bộ Y tế, Bộ Giáo dục và Đào tạo (1998),Bài giảng dược liệu tập II, Ha Noi.
2. Hải Thượng Lãn Ông, Thuốc Việt Nam, Nhà xuất bản Văn hóa dân tộc.
3. Phạm Trương Thị Thọ (1996), “Điều kiện tối ưu chiết xuất eugenol từ tinh
dầu hương nhu” ,Tạp chí khoa học và cơng nghệ , (2), pp 28 -33.
4. Giang Thị Sơn (1996) , “Điều chế eugenol có độ tinh khiết cao làm nguyên
liệu bán tổng hợp các dãn chất của eugenol”, Tạp chí dược học, (5), pp. 8-9.
5. Nguyễn Thị Hoàng Oanh, Tách eugenol từ tinh dầu hương nhu trắng và
chuyển hoá thành một số sản phẩm hương liệu ,Graduation essay , Vietnam
National University, Hanoi.
6. Phạm Trương Thị Thọ (1994), Nghiên cứu bán tổng hợp định hướng các dẫn
xuất eugenol có tác dụng kháng nấm Candida albican , Science doctoral
thesis, Hanoi.

-

References English
7. 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”.
8. 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”.
9. 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, 9 (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