*

ESSENTIAL OIL MICROENCAPSULATION WITHIN ALGINATE:

FORMULATION AND EVALUATION OF ANTIFUNGAL ACTIVITY

UNDER GUIDANCE OF
MR. RAIBARINDER SINGH
Assistant Prof. (P’ceutics)
PREPARED BY
JATINDER VERMA
ROLL NO. 1502001041
 *
*INTRODUCTION
*OBJECTIVE & PLAN OF WORK
*LITERATURE REVIEW
*MATERIAL & METHODS
*RESULTS AND DISCUSSION
*JUSTIFICATION
 *
* Essential oils (EOs) are the volatile lipophilic components extracted
from plants.
* Many essential oils (EOs), such as garlic, cinnamon, thyme, oregano,
clove, basil, coriander, citrus peel, eucalyptus, ginger, rosemary, and
peppermint, among others, have been demonstrated antimicrobial
activity.
* The biological activity of EOs can be lost by volatilization of active
components or its degradation by act of high temperature, oxidation
and UV light.
* These disadvantages make the commercial application of these oils
limited.
* Micro- encapsulation as one of the most efficient methods of
formulation by which solids, liquids or even gases have the desired
characteristics in extremely tiny amounts may be enclosed in
microscopic particles formation of thin coatings of wall material with
limited permeability around the substances
 *Introduction (cont….)
* The commercial applications of these EOs require a suitable
formulation constituted by biodegradable compounds that
protect them from degradation and evaporation at the same
time that allows for a sustained release.
* Via microencapsulation to reduce the rate of evaporation of
the oil.
* Micronapsulation in Calcium alginate microspheres may
effectively reduce the evaporation rate of essential oils, thus
increase the potential antifungal activity .
 *

* The ability to incorporate reasonably high

concentrations of the drug.
* Stability of the preparation after synthesis with a
clinically acceptable shelf life.
* Controlled particle size and dispensability in
aqueous vehicles for injection.
* Release of active reagent with a good control over a
wide time scale.
* Biocompatibility with a controllable
biodegradability and
* Susceptibility to chemical
 *

* The aim of the present

study is to develop
microencapsulation of
clove oils in calcium
alginate by emulsion
extrusion technology in
order to achieve their
optimal antifungal
activity against
pathogenic fungi species
Aspergillus niger.
 *

* Extraction of volatile oil from clove by Clevenger

Apparatus.
* Microencapsulation of essential oils by alginate.
* Formation of fungal culture of aspergillus Niger.
* Evaluation of microcapsules antifungal activity by
the culture media.
 *

* ESSENTIAL OIL DATA

 Name: CLOVE
 Biological source: Cloves are the aromatic flower buds of
Syzygium aromaticum from family Myrtacecace.
* Collection and preparation: The flower buds are collected when
their lower part turns from green to crimson. The collection
takes place twice year in August and December. The
inflorescences are collected from movable platform. The cloves
are dried in open air and in mats and separated from their
peduncles, latter forming separate article of commerce known as
“clove stalks” If left to long on the tree, the buds open and the
petals fall, leaving” blown cloves” later the fruits known as”
mother cloves” are produced a proportion of these, usually at a
stage intermediate between that of a clove and a fully riped
fruit, are frequently found in a drug. Cloves are imported in bales
covered matting made of strips of coconut leaves.
* Microscopically character: Cloves are 10-17.5mm long.
* The stock of clove consists of cylindrical hypanthium or swelling
of the torus and above which is a bilocular ovary containing
numerous ovules attached to axile placentae. Consist of four
slightly projecting calyx teeth; four membranous imbricated
petals and numerous in curve stamens around large style.
* Cloves have strong, fragment, and spicy order and pungent,
aromatic taste. When indented with finger nail, they readily
exude oil. Cloves sink in freshly boiled and cold water.
* Uses: Cloves as used as a stimulant aromatic and a spice.
* The preparation of volatile oil is used as an antifungal
preparation.
 CLOVE OIL
* Colour: It is colourless or pale yellow liquid.
* Charactersites : It is slightly heavier than water (relative
density1.047 to 1.060). It is soluble in from 1 to 2 volume
alcohol.
* Chemical constituents: Clove oil contains 84 to 95% of phenols
(eugenol with about 3% of acyl ugnol) sesquiterpenes and small
quantities of ester, ketones and alcohols.
 *

* FOR EXTRACTION OF ESSENTIAL OIL:

* Appratus used : cleveger apparatus

Sr.no Ingredients Quantity

1 Cloves(powdered) 50g

2 Distilled water 250 ml

* MICROENCAPSULATION MATERIAL:
* Apparatus used: Beakers, stirrer ,syringe
Sr. no. Ingredients Quantity

1 Clove oil 10ml

2 Sodium alginate 10gm
3 Hexane 5ml
4 Calcium chloride -
5

* MICROBIAL STRAIN PREPRATION

* Apparatus used: beakers, petri dish,incubaor ,autoclave
sr. no. Ingredient Quantity

1 Peptone 10gm

2 Dextrose 40gm

3 Agar 15gm
 *
* Essential oil extraction by hydro-distillation
Dried flower buds of Cloves plant was taken

400 g of dry cloves were powdered

Then immersed them in a 250 ml flask with distilled water

for 3-4 hours.

Essential oils recovered in small opaque bottles

* Microencapsulation method

sodium alginate was dissolved in distilled water to produce alginate

solutions

sodium alginate solution (30 g) and EO (10 g) were homogenized

into a 200 mL beaker

oil was added to the alginate solution during mixing until the desired
oil loading was obtained.50 ml of alginate-oil emulsion was then sprayed
into a collecting water bath containing calcium chloride solution

resulting microcapsules were allowed to harden in the CaCl2 solution

oil-loaded alginate beads were collected from the cross-linking solution

using a sieve.
* Microbial culture
Suspend 65 g of the medium in one liter of purified water

Heat with frequent agitation and boil for 1 min to completely dissolve
the medium

Autoclave at 121° C for 15 minutes

Cool to 45 to 50°C and pour into petri dishes or tubes for slants

For processing of specimen, streak the specimen onto the medium

with a sterile inoculating loop in order to obtain isolated colonies

Incubate the plates at 25 – 30°C in an inverted position (agar side up)

with increased humidity.
 *

* ESSENTIAL OIL
* R%=M*100/M0
* Whereas
R (%): efficiency
M: weigh of extract
recovered in g.
M0: weigh of dry extract
used for the extraction in g

* R%=173*100/400
* =43.25%
*
*
 *

The final result I got while performing preparation

and evaluation of antifungal activity of essential oil
microcapsule is evaluated
 *
1. The result indicated that the encapsulation process of tested oil
can be considered as inexpensive and efficient technique to
maintain the anti- fungal activity of the oils.
2. It promote the ease of handling of the oils, where the
microencapsulated oil clove exhibited high growth inhibition for
Aspergillus niger
3. The results obtained from this study suggest that of all
formulation techniques, microencapsulation seems to be the best
choice for increasing the use of EOs.
4. The microencapsulation in alginate is expected to be suitable
for other hydrophobic essential oils.
LEARNINGS
While performing the project I came to knew about;
1. I came to know about extraction of essential oils
2. Making of microcapsules of alginate with essential oil of clove
3. Formation of fungal strain by extraction spores from garlic.
 *

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