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AQUEOUS EXTRACTION OF EUGENIA CARYOPHYLLUS BUDS

(CLOVE) FOR ANTI -TOOTH DECAY

P. Amani,* S. Bharathbhushan Reddy, G. Karthik Reddy Ashwini, M. Rajeshwari,

Ragya E.

Venkateshwara Institute of Pharaceutiical Sciences, Cherlapally, Nalgonda, Telangana.

ABSTRACT

A cohort of patients with overgrowth syndromes has been identified

with congenital lipomatous overgrowth, deregulated fat deposits, and

mixed vascular malformations. The acronym CLOVES was given on a

heuristic basis to stand for congenital lipomatous overgrowth (CLO),

vascular malformation (V), epidermal nevi (E), and scoliosis and

spinal deformities (S). These patients have upper limb anomalies with

variable phenotypes. Although hand anomalies alone cannot make the

diagnosis, the foot, truncal, cutaneous and spinal anomalies are

particularly diagnostic. clove syndrome has emerged as a distinct

clinical entity diagnosed by clinical and radiographic examinations.

The overgrowth pattern is now easily distinguished from other overgrowth syndromes.

Toothpaste is a paste or gel to be used with a toothbrush to maintain and improve oral health

and aesthetics. Since their introduction several thousand years ago, toothpaste formulations

have evolved considerably - from suspensions of crushed egg shells or ashes to complex

formulations with often more than 20 ingredients. Among these can be compounds to combat

dental caries, gum disease, malodor, calculus, erosion and dentin hypersensitivity.

Furthermore, toothpastes contain abrasives to clean and whiten teeth, flavors for the purpose

of breath freshening and dyes for better visual appeal.

1. INTRODUCTION

Medicinal plants have been used in traditional healing practices for treating various human

ailments since time immemorial. Such traditional have provided the basis of scientific

investigation on medicinal plants which led to the discovery of many potential drug

molecules of alternative medicine for treating human disorders around the world. In recent

*Corresponding Author

P. Amani

Venkateshwara Institute of

Pharaceutiical Sciences,

Cherlapally, Nalgonda,

Telangana.

Article Received on

24 Nov. 2016.

Revised on 14 Dec. 2016,

Accepted on 04 Jan. 2017

DOI: 10.20959/wjpps20172-8531

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 6.647

Volume 6, Issue 02, 767-778 Research Article ISSN 2278 – 4357

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Amani et al. World Journal of Pharmacy and Pharmaceutical Sciences

years, there has been a dramatic rise in use of herbal drugs/preparations in the developed

countries because of their easy availability and cost effectiveness besides having desired

pharmacological effectiveness with high level of safety/ low toxicity profile. It is estimated

that world’s one-fourth population i.e. 1.42 billion people are dependent on traditional herbal

medicines for the treatment of various ailments. However, the lack of documentation and

stringent quality control procedures has hindered the easy acceptance of such plant drugs

(crude preparations) to be used as herbal medicine. According to the WHO, to ensure

reproducible quality of herbal plants (or preparations), physicochemical and photochemical

characterizations are required to be carried out for establishing their identity, purity, and

quality standards Therefore, there is a need for documentation of standardization studies for

profiling the quality control parameters of plants-derived crude/herbal drugs.

Plant image

Eugenia Caryophyllum (CLOVE)

Kingdom : Planate

Order : slightly aromatic

Family : myrtaceae

Genus : Eugenia

Species : caryophyllum

Biological source:

It consists of the dried flower buds of Eugenia caryophyllus.

Family: Myrtaceae.

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Macroscopical characters

I. Size: Length varies from 12 to 17 mm.

II. Type: Actinomorphic, bisexual, epigynous. The flower bud has a spherical head and a

sub-cylindrical hypanthium tapering at the lower end.

III. Calyx: Polysepalous, 4 hard and thick sepals with oil glands

IV. Corolla: Polypaptalous, 4 petals imbricate, enclose the stamens and forms the head of

the bud Androecium- numerous stamens, free and introrsely;

V. Gymnasium: Binocular, inferior with ovules stamens, free Placentation axial.

VI. Style: Single and erect.

VII. Colour: Dark brown;

VIII. Odour: Aromatic, spicy, Strong.

IX. Taste: Pungent, aromatic.

X. The volatile oil is situated in the schizolysigenous oil glands or ducts which are

present in all 1 parts of the flower buds (hypodermis).

Microscopical characters of Clove flower bud

Transverse section of clove hypanthium below the ovary shows epidermis, cortex and

columella

1. Epidermis: Single layered small cells with straight walls and has a very thick cuticle.

Epidermal layer gets intercepted by Ranunculaceous type of stomata.

2. Cortex: The three distinct zones or regions in the cortex can be made out.

(a) The peripheral region containing 2 to 3 layers of big, ellipsoidal, schizo-lysigenous oil

glands embedded in the radially elongated parenchymatous cells.

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(b) The middle region containing 1 or 2 rings of bicollateral vascular bundles associated with

a few pericyclic fibres, embedded in thick walled parenchyma and

(c) The inner region made of loosely arranged aerenchyma.

3. Columella: Forms the central cylinder containing thick walled parenchyma with a ring of

bicollateral vascular bundles towards the periphery of the cylinder. Numerous sphaeraphides

are seen scattered throughout the columella and to a certain extent in the middle cortical zone.

Section cutting of clove bud

Chemical constituents

1. Volatile oil (16-21%):- Phenol chiefly Eugenol (80-88), acetyl eugenol (10- 15%); α and

β -Caryophyllene.

2. Tannins (10-13%) – Pyrogallol tannins.

3. Other substances are methyl furfural and dimethyl furfural.

Uses

1. Antiseptic.2. Stimulant.3. Carminative.4. Flavoring agent.5. Local anesthetic (Eugenol).6.

Spice 7. Used in toothache, dental preparations and mouthwashes.

8. Oil in perfumery.9. Clove oil and zinc oxide are used in temporary filling of dental

cavities.

MATERIALS AND METHODS

Collection

Inflorescence of clove is panicle or compound raceme and branches are opposite and

decussate. Clove buds are at first white, then green and finally become crimson-red in colour.

Collection of crimson-red coloured buds carried out in the dry weather from August to

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December. For collection of buds, natives climb the trees or put ladders and pick the buds,

with the stalk. Mobile platforms are also used for collection. Trees are also beaten by means

of bamboos sticks; clove buds fall on the ground and are collected. After collection stalks are

separated and then are put on coconut mats or concrete floors and dried in sun. During night

they are covered. Drying takes about three days. As a result of drying clove buds become

dark reddish-brown and loss about 70% of the weight. Clove is then graded according to size,

Condition and quality, packed into bales and exported.

Chemicals and Reagents

Methanol, Acetone, Sudan red III, water chemical in laboratory in used the clove.

Tragacanth, glycerin, magnesium carbonate, magnesium hydroxide, sodium lauryl sulphate,

flavoring agent (clove oil), vehicle (water), sorbitol dextrose.

Equipment’s required

Clevenger’s , heating mantle , beakers, mortar priest , petriplates , conical flask ,electronic

balance , fluorescence analysis cabin , round bottom flask ,pH meter , slides, stirrer, eggshell,

toothbrush.

3. EXTRACTION OF EUGENOL FROM CLOVES

EXPERIMENTAL PROCEDURES

Steam distillation

Set up a distillation apparatus using a 500-mL round-bottom flask. Use a Claisen as well as a

regular distillation head. You will use the main arm of the Claisen head to add water. Attach

the take-off adapter, securing it with a rubber band. Set the ring with asbestos gauze high

enough that you can control the amount of heat easily. Use a Fisher burner if possible.

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Mount the receiving flask onto a stand using a utility clamp. Use a tripod or a large ring

clamp and wire gauze to position the ice-water bath. Use a steam-bath bowl or 1000-mL

beaker for ice water.* At least half of the receiving flask should be immersed in ice water.

Weigh 14–16 g of cloves using a 100-mL beaker. Crack or squash them somewhat so water

can penetrate them better. Weigh again and record the exact mass.

Place the cloves in the round-bottom flask. Pour 150 mL of distilled water and drop in 3

boiling sticks.† Note the level of water.

Place a 250-mL separatory funnel with a joint in the main arm of the Claisen adapter,

inserting it through a ring clamp. Make sure the stopcock is closed and pour in about 200 mL

of distilled water. If a separatory fun-nel is not available, close the Claisen head with a glas

Teflon stopper.

Turn on cold water to flow briskly through the condenser and slowly bring the mixture to

boiling. Note that it will foam while boiling. Be careful when it just begins to boil or the foam

may burst and get into the con-denser and contaminate the distillate. Once the mixture is

boiling evenly, increase the heat.

Distill the mixture, making sure that it boils rapidly but that foam does not get into the

condenser. Foam should completely fill the round-bottom flask during the distillation.

Collect the milky distillate into a 250-mL Erlenmeyer flask standing in ice water. If a clear

distillate is being collected, you are distilling just water.

Continue distilling until no more oily material can be seen in the con-denser (it will take at

least 45–60 minutes). You should collect at least 150–200 mL, but no more than 225 mL, of

distillate.

Remove the clove residue from the flask while it is still warm, or at least fill the flask with

tap water. To remove the solids fill the flask with tap water, swirl, pour the mixture into a

1000-mL beaker, decant liquid into the sink, and then put the solid residue into the trash (do

not put the solid residue into the sink as it may clog the drain).

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TOOTHPASTE

4.1 INTRODUCTION

Toothpastes claim to clean, remove plaque, prevent cavities, and whiten teeth. Is there really

a difference in toothpastes Is one truly superior to another Does a higher price mean tooth

paste is better Teeth are the calcified structures in our mouths that are used to break down

food. Teeth are made of multiple tissues of varying density and hardness.

The two major parts of a tooth are the crown and the root. The roots of teeth are embedded in

the maxilla (upper jaw) or the mandible (lower jaw) and are covered by our gums. The crown

is the visible portion that extends above the gums.

Table 1. Components of toothpaste

Ingredients % by

weight

Tragacanth 1.0gm

Glycerin 31.0gm

Magnesium carbonate 1.0gm

Magnesium hydroxide 3.0gm

Sodium lauryl sulphate 1.0gm

Water q.s

Calcium carbonate 44.5gm

Clove oil 2gm

Table 2. Typical toothpaste ingredients

Gums Inorganic

Abrasives Surfactants Humectants Tartar Control

Thickeners Ingredient

Sodium Sodium lauryl

Glycerin Tetra sodium

carboxym

ethyl

Silica

thickeners Hydrated silica

sulfate pyrophosphate

cellulose

Sodium Dicalcium Sodium N-

lauryl

Cellulose

ethers aluminum phosphate Sorbitol Gantrez S-70

sarcosinate

silicates dehydrate

Xanthan

Gum Clays

Calcium Pluronics

Propylene

glycol

Sodium tri-

carbonate polyphosphate

Carrageen Sodium Xylitol

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ans bicarbonate

Sodium

alginate

Calcium Sodium lauryl Polyethylene

pyrophosphate sulfoacetate glycol

Carbopols Alumina

Some Active Toothpaste Ingredients are

Tartar control additives, such as tetra sodium pyrophosphate, do not remove tartar already

formed on your teeth; they react with the minerals in your saliva to prevent them from

forming tartar.

Baking soda, sodium bicarbonate, is a mild abrasive and reduces stains on the teeth. It

neutralizes acids in the saliva and provides a foaming action in the mouth.

Triclosan (2,4,4’–trichloro-2’-hydroxydiphenyl ether) is a chlorinated aromatic compound

with functional groups that include both phenols and ethers. It is an antibacterial additive that

clings to your teeth after brushing and continues to kill bacteria

Fluoride is effective in strengthening tooth enamel. If the toothpaste is approved by the

American Dental Association, then the fluoride is effective. Differences in the amount of

fluoride has little effect.

Whitening ingredients do little in your toothpaste other than remove some stains. (All

toothpastes remove stains.) No toothpaste can whiten your teeth permanently.

Gels vs. pastes are more of a personal preference. Gels use silica as an abrasive to polish

teeth while pastes usually use calcium carbonate.

Non-Active Ingredients Include

Sodium lauryl sulfate and ammonium lauryl sulfate are surfactants that promote foaming

action. The foaming action is equated with cleaning. Alginate or Xanthan gum is binding

agents to maintain the consistency of toothpaste for mouth feel properties.

Sodium benzoate or ethylparaben are preservatives to prevent growth of micro-organisms.

Humectants, such as water, orbital or glycerin, hold moisture so the toothpaste does not dry

up Flavoring is added to impart a pleasant taste. Sodium saccharine is added to give the

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toothpaste a sweet taste. Titanium dioxide is considered to be a coloring agent. It makes the

toothpaste opaque and gives it a white color.

Artificial dyes are added to make colored toothpastes such as blue, green or red.

In this experiment, we will make 3 simple toothpaste formulations and evaluate them, along

with some commercial toothpaste for its properties and cleaning effectiveness.

Evaluation parameters

Preparation of Toothpaste Solutions For Testing

Weigh 2.0 g of toothpaste into a 250-mL beaker. Add 80 mL of water to the beaker. Stir to

form a suspension. The mixture of toothpaste and water may have to sit for up to 30 minutes

in order to get a uniform suspension.

Determination of Hard and Sharp Edged Abrasive Particles

Squeeze a piece of toothpaste (homemade or commercial) approximately 3 cm long onto a

piece of waxed paper.

Using your finger, press it along its length to test for the presence of hard and sharp edged

abrasive particles

Abrasiveness

Place a pea-sized amount of toothpaste (homemade or commercial) on a clean plastic

microscope slide.

Add 1 or two drops of distilled water to the toothpaste sample.

Using a clean cotton swab, rub the toothpaste sample in a back and forth motion 25 times

using short 1 cm strokes.

Carefully rinse off the microscope slide and dry it with a soft tissue or paper towel.

Examine the slide under a dissecting microscope, illuminated from above, and determine the

amount of scratches on the surface of the slide. Rate the scratches on a scale from 0 (no

scratches) to 5 (a high degree of scratches).

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Determination of Spread Ability

Measure 1 g of toothpaste sample (homemade or commercial). Place the sample at the center

of a glass plate (10 x 10 cm). Place a second glass plate over the sample.

Place a 1 kg weight on top of the glass plate. Be careful to avoid sliding of the plate.

After 10 minutes, remove the weight and measure the diameter of the paste in centimeters

Determination of Ph

Pour about 40 mL of the toothpaste solutions prepared in step A, above, into clean 100 mL

beakers. We observed the pH

value 11.86

Measure the pH of each solution using a pH meter.

Determination of Foaming Ability

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Pour 30 mL of one of the toothpaste solutions prepared in step A, above, into a 100 mL

graduated cylinder.

Cover the top of the cylinder with a piece of Para film. Place your hand over the Para film

and shake the graduated cylinder 25 times.

Place the graduated cylinder on the lab bench and measure the height of the foam above the

water in cm. The foaming ability is the height of the foam.

CONCLUSION

Toothpaste was made long ago. The most important ingredient in toothpaste is fluoride.

There are lots of chemicals in toothpaste. The texture, fragrance, and color helps the

toothpaste, because without these things toothpaste wouldn't be as pleasing. If we didn't have

toothpaste, we would have bacteria and plaque. Our teeth would not be as strong and teeth

would decay. So, you should be thankful because without toothpaste our breath wouldn't be

as fresh as it is now.

RESULTS

Macroscopic Characters

Colour Crimson of dark brown

Taste Aromatic

odour Pungent and aromatic

Identication test Results

Preparation of toothpaste solution for testing +ve

Abrasiveness +ve

Determination of spread ability +ve

Determination of pH +ve

Cleaning ability +ve

DISCUSSIONS

The quantitative determination of pharmacognostical parameters is useful for setting standard

for crude drugs.Macroscopic studies play an important role for primary identification of

drugs. Whitening ingredients do little in your toothpaste other than remove from stains.as per

per evaluation parameter we checked all details of clove paste and determine the

spreadability property,foam property,abrasive effect of dental for bright ness of tooths.

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