HIMA HARIDASAN

Dept. Of Microbiology,

St. Mary’s college,

Thrissur- 680020,

University of Calicut

E-mail: himaharidasan93@gmail.com

Family : Rutaceae

Native to India.

Leaves - Indian diet to improve

appetite and digestion.

Deciduous to semi-evergreen

aromatic tree

TRADITIONAL USES

Analgesic - pain-killing

Febrifuge - reduce fever

Stomachic - improves

appetite and digestion

Carminative - relieves

flatulence

Treatment of dysentery

& skin eruptions

PROPERTIES

Anti-oxidative –inhibits oxidation damages

Cytotoxic – cell-toxic

Antimicrobial

Anti-ulcer

Positive inotropic - modifies the force or speed of muscle contraction

Cholesterol reducing activities

1)Preliminary phytochemical analysis

2)Antimicrobial analysis

MATERIALS AND METHODS

1)PLANT MATERIAL

2)PREPARATION OF EXTRACT

a) The powdered leaves (30g) - methanol extraction

b) Filteration - whatman filter paper

c) Filtrate – evaporation (65ºc)

d) Organic semisolid mass was dried

The bacterial cultures: –

o Gram positive -Staphylococcus and Bacillus

o Gram negative - Escherichia coli, Pseudomonas, Proteus and Klebsiella (obtained from Poly clinic, Thrissur).

o Were grown and maintained in nutrient broth medium at 37ºc.

The fungal cultures :-

o Aspergillus flavus, Alternaria

and Pencillium ( isolated by

air sampling in Microbiology

laboratory, St.Mary’s college,

Thrissur).

o Were maintained on

sabourauds dextrose agar

(SDA) slants.

1.Detection of Tannins

By Ferric chloride test –

extract + distilled water

(5mg) (5ml)

mixture neutral 5%

feCl3 soln

added

blue green color

(presence of tanin)

2.Detection of Phenols

By Lead acetate test –

extract + distilled water

(5mg) (5ml)

10% Pb acetate

soln (3ml) added

bulky white precipitates

(presence of phenols)

3. Detection of flavanoids

The chloroform extract of the

solution

Treated with

NH4OH soln

Yellow fluorescence

(presence of flavanoids)

4. Detection of Terpenoids

Chloroform (2ml) + conc.

H2SO4

added carefully to

0.5ml of extract

red brown color at the

interface

(presence of terpenoid)

5. Detection of Quinone

Conc. H2SO4 (1ml)

added to

plant extract (1ml)

Red colour

(presence of Quinones)

6. Detection of steroids and

phytosteroids

Plant exract + chloroform

(0.5 ml) (0.5 ml)

added few drops of

conc. H2SO4

brown ring / bluish- brown

ring(steroids) (phytosteroids)

7. Detection of carbohydrate

Molisch’s test –

Molisch’s reagent + extract solution

(2 drops) (1 ml)

mixed well

Inclined the tube

Added conc. H2SO4(1ml) along the sides of the tube

Reddish violet ring at the junction of two liquids

(presence of carbohydrate)

8. Detection of Proteins

The extract + distilled water

(10ml)filtered through

Whattmann No.1 filter paper

filtrate

Used to tests for proteins and

aminoacids

8.a) Biuret test –Filtrate + 2% CUSO4 soln

(aliquot) (a drop)

added

95% ethanol(1ml)

added

excess of potassium

hydroxide pellets

The pink color in ethanol layer

(presence of proteins)

8.b) Detection of aminoacids

Ninhydrin test –

Ninhydrin solution + Filtrate

(2 drops) (2 ml)

Purple colour(presence of amino acids)

9. Detection of phytosterolsLibermann-Burchard’s test –

Extract +dry chloroform

Dissolved in a dry test tube

Acetic anhydride (several drops) was added

Conc.H2SO4 (2 drops) was added

Mixed carefully

After the reaction finished

Cholesterol conc. was measured using spectrophotometry

Method : Well diffusion Assay

Procedure :-

Swabbing

Short incubation

Wells cutting

100µg, 200µg, 300µg and 400µg of 0.5g leaf extract prepared in 1000ml of methanol – loaded.

Centre well- 100µl methanol, control.

Incubation

Zone of inhibition diameter - recorded.

1)PHYTOCHEMICAL SCREENING

SL.NO. PHYTOCHEMICALS OBSERVATION INFERENCE

1 Tanin blue green color Present

2 Phenol bulky white

precipitate

Present

3 Flavanoid yellow fluorescence Present

4 Terpenoids red brown colour at

the interface

Present

5 Quinone red colour Present

SL.NO PHYTOCHEMICALS OBSERVATION INFERENCE

6 Steroid &

phytosteroid

brown and bluish

brown ring

Present

7 Carbohydrate reddish violet ring Present

8 Protein pink colour Present

9 Amino acid purple colour Present

10 Phytosterols No deep green colour Absent

Phytochemical compounds in the methanolic extract of Murraya koiengii leaves

Sl.

No.

TEST

MICROORGANISM

ZONE OF

INHIBITION

1 Staphylococcus Present

2 Bacillus Present

3 Proteus Present

4 Escherichia coli Absent

5 Pseudomonas Present

6 Klebsiella Absent

ZONE OF INHIBITION (mm) FOR DIFFERENT QUANTITY OF EXTRACT

100µg 200 µg 300 µg 400 µg

20mm 25 mm 33 mm 40 mm

_ 14mm 16mm 21mm

_ 14mm 25mm 34mm

_ _ _ _

_ 20mm 25mm 30mm

_ _ _ _

Antibacterial efficiency of methanolic extracts of Murraya koiengii leaves.

Staphylococcus species inhibited by methanolicextracts of Murrayakoiengii leaves.

The three Gram positive bacteria showed sensitivity against the methanolic extract

Staphylococcus sp., emerged as the most susceptible.

Among the fungal strains, none of the tested ones showed considerable inhibition by the methanolic extract.

So, Murraya koenigii is a common remedy among the

various ayurvedic practitioners for treatment of

diversity of ailments.

So, it is used in foods.

Hippocrates, the father of medicine proclaimed

“Let food be thy medicine and medicine be thy food”.

THANK

YOU