ANTIMICROBIAL ACTIVITY AND PHYTOCHEMICAL SCREENING OF CRUDE EXTRACT OF zaleya decandra (L).Verm

1. ANTIMICROBIAL ACTIVITY AND PHYTOCHEMICAL
SCREENING OF CRUDE EXTRACT OF zaleya decandra
(L).Verm
BY
KARTHICKSAMY.S
KAVIARASU.E
KAVIN.V
KRISHNA JAYANDHA.S
MAHENDRA PRASANTH.M
UNDER THE GUIDANCE OF
S.SRI BUVANESHWARUI.,M.Pharm.,
ASSOCIATE PROFESSOR.M,
DEPARTMENT OF PHARMACEAUTICAL
BIOTECHNOLOGY

2. INTRODUCTION
ZALEYA DECANDRA;
• The plant Zaleya decandra is commonly called as Purslane (English)
belongs to family aizoaceae. It is an annual branched, succulent herb in
India
• Zaleya decandra and its species are used for antidiabetic [4], anti-
inflammatory,
• Antihyperglycemic, hepatoprotective, antioxidant and diuretic.
• Medicinal plants are a source of great economic value in Indian continent.
• Nature is an important source of medicines. Medicinal plants constitute
main sources of pharmaceuticals and health careproducts and
nutraceuticals.
• Many ancient nations awakened to the importance of herbal medicine
which brings more cures.

3. PLANT PROFILE
• Botanical name : Zaleyadecandra
• Synonyms :Zaliadecandra, zeleijadecandra
• Family : Aizoaceae

4. Medicinal Uses:
1. It has wide tradiotional medicinal uses.
2. Root of this plant used for treatment of Hepatitis,Asthma,
Orchitis.
3. Leaves (juice) dropped in nostril to relieve partial headache.
4. Flavonoids shows Anti-Inflammatory activity.
5. Curative properties – Treatment of various ailments such as
Burns , Wounds ,Fever , tooth ache ,Skin disease, Anti-
diabetic,Stomach disease ,Respiratory tract infection &
cough.

5. PREPARATION OF PLANT EXTRACT:
PROCEDURE:
SOXHLET METHOD OF ETHANOL EXTRACTION;
Plant material can be fresh (for example, a plant leaf) or dried. It
needs to be crushed, using a pestle and mortar, to provide a greater
surface area. The plant material should be sufficient to fill the
porous cellulose thimble (in our experiments we use an average of
14 g of thyme in a 25- x 80-mm thimble). All equipment should be
provided for students to assemble. Allowing students to build the
extraction apparatus may give them a greater appreciation for the
process of extraction, as opposed to testing an antimicrobial
compound out of a purchased bottle. The students should begin by
building a rig using stands and clamps to support the extraction
apparatus. Following this, the solvent (150 ml of ethanol) is added
to a round bottom flask, which is attached to a Soxhlet extractor .

6. • The crushed plant material is loaded into the thimble, which is placed
inside the Soxhlet extractor. The side arm is lagged with glass wool. The
solvent is heated using the isomantle and will begin to evaporate, moving
through the apparatus to the condenser. The condensate then drips into the
reservoir containing the thimble.
• Once the level of solvent reaches the siphon it pours back into the flask
and the cycle begins again. The process should run for a total of 10 hours.
Once the student has set up the extraction it can be left to run without
direct supervision. It is not advised to leave the equipment completely
alone due to the mix of running water and an electrical appliance, so a
technician or other lab user should be made aware.

7. • The equipment can beturned on and off when overnight running is not
permitted, and the time split over a number of days. For good practice, a
control should be added. This could be plant material that has no known
antimicrobial effect (for example, a carrier oil such as sunflower oil) at the
testing stage. Once the process has finished, the ethanol should be
evaporated using a rotary evaporator, leaving a small yield of extracted
plant material (about 2 to 3 ml) in the glass bottom flask.
PHYTOCHEMICAL SCREENING:
The prepared extract of the Zaleya decandra plants was used to test
various phytoconstituents present in them. Different chemical reagents
were prepared and specific test, for specific phytochemicals was done.
These various tests were qualitative and hence termed phytochemical
screening. All chemicals and solvents were procured from Fisher Scientific,
India, and were used without further purification.

8. EXPERIMENT OBSERVATION INFERENCE
Mayer’s Test:Filtrates
were treated with Mayer’s
reagent (Potassium
Mercuric Iodide).
Formation of a yellow
coloured precipitate.
Presence of alkaloids.
Molisch’s Test: Filtrates
were treated with 2 drops
of alcoholic α-naphthol
solution in a test tube.
Formation of the violet ring
at the junction.
Presence of
Carbohydrates.
Modified Borntrager’s
Test: Extracts were
treated with Ferric
Chloride solution and
immersed in boiling water
for about 5 minutes. The
mixture was cooled and
extracted with equal
volumes of benzene. The
benzene layer was
separated and treated
with ammonia solution.
Formation of rose-pink
colour in the ammoniacal
layer.
Presence of anthranol
glycosides.

9. EXPERIMENT OBSERVATION INFERENCE
Ferric Chloride Test:
Extracts were treated with
3-4 drops of ferric chloride
solution.
Formation of bluish black
colour.
Presence of phenols.
Gelatin Test: To the extract,
1% gelatin solution
containing sodium chloride
was added.
Formation of white
precipitate.
Presence of tannins.
Alkaline Reagent Test:
Extracts were treated with
few drops of sodium
hydroxide solution.
Formation of intense
yellow colour, which
becomes colourless on
addition of dilute acid.
Presence of flavonoids.

10. ANTI MICROBIALASSAY;
Preparation of Nutrient Agar;
• Suspend 28 g of nutrient agar powder in 1 litre of distilled water.
• Heat this mixture while stirring to fully dissolve all components.
• Autoclave the dissolved mixture at 121 degrees Celsius for 15 minutes.
• Once the nutrient agar has been autoclaved, allow it to cool but not
solidify.
• Pour nutrient agar into each plate and leave plates on the sterile surface
until the agar has solidified.
• Replace the lid of each Petri dish and store the plates in a refrigerator.

11. AGAR WELL DIFFUSION METHOD ;
• Agar wel ldiffusion method is widely used to evaluate the an-timicrobial
activity of plants or microbial extracts .
• Similarly to the procedure used in disk-diffusion method,thea gar plate
surface is inoculated by spreading a volume of the microbialin-oculum over
the entire agar surface.
• Then,a hole with a diameter of 6 to 8mm is punched aseptically with a
sterile corkborer or a tip, and a volume (20–100 mL) of the antimicrobial
agent or extract solution at desired concentration is introduced in to the
well.
• Then,agar plates are incubated undersuitable conditions depending upon
the test microorganism.
• The antimicrobial agent diffusesin the agar medium and inhibits the
growth of the microbial strain tested.

12. RESULT AND DISCUSSION;

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79. https://doi.org/10.1016/j.phymed.2020.153343, Accessed: 04.11.2020.
4.Ballard, C.R., & Marostica, M.R. (2019). Health benefits of flavonoid., in
book Bioactive Compounds, 185-201. https://doi.org/10.1016/b978-0-12-
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