EVALUATION OF ANTIFUNGAL POTENTIAL OF LEUCAS

LAVANDULIFOLIA

SUBMITTED BY: PRATIK SWARUP DAS Regd. No – 072331 of

2012-2013 Roll. No – BP/ 12/42

Under the guidance of Mrs. SMRITI REKHA CHANDA DAS

(M. Pharm), Asst.Professor, Dept. Of pharmaceutical chemistry (GIPS)

PRESENTATION ON

The aim and objective of this project work is to evaluate anti-fungal potential of aerial part of Leucas lavandulifolia on benzene and acetone extracts.

AIM & OBJECTIVE

AUTENTICATION OF PLANT

 COLLECTION OF PLANT MATERIAL: (AERIAL PARTS)

  Leucas lavandulifolia were collected from Azara,

Hatkhowapara of Guwahati (Assam). Washed properly washed in tap water and

rinsed in distilled water Dried in oven at a temperature of 35 – 400C for

3 days and powdered . Stored in air tight glass containers, protected

from sunlight until required for analysis.

EXPERIMENTAL WORK

 Method:   Preparation of benzene/acetone extracts 100g of powdered samples in 400ml

petroleum ether and kept for 3 days . powder samples were dried from petroleum

ether and dried in room temperature and soaked the powdered samples in 500ml of benzene/acetone and macerated for 7 days .

The extract was then filtered using filter paper then concentrated to 50ml by using rotary evaporator and stored in airtight container.

Extraction of plant material

Observations

BE: Benzene extract; AE: Acetone extract; ‘+’: presence of phytochemical; ‘-’: absence of phytochemical. 

PHYTOCHEMICAL SCREENING

PHYTOCHEMICAL

CONSTITUENTS

L. Lavandulifolia

BE AE

ALKALOIDS + +

CARBOHYDRATES - +

GLYCOSIDES + -

SAPONINS + +

PHYTOSTEROLS - +

PHENOLS + -

TANINS + +

FLAVANOIDS + +

PROTIENS - -

AMINO ACIDS - -

.

FUNGAL STRAIN Trichophyton rubrum & Microsporum fulvum. STANDARD DRUG 1% solution of miconazole nitrate.COMPOSITION OF SABOURAUD DEXTROSE AGAR (SDA):

ANTIFUNGAL ACTIVITY 

Ingredients Gm/L

Mycological peptone (enzymatic digest of casein and animal tissues) 10 gm

Dextrose 40 gm

Agar 15 gm

pH adjust to 5.6 at 250 C

Suspend 65 g of the medium in one liter of purified water.Heat with frequent agitation and boil for one minute to completely dissolve the medium.Autoclave at 121° C for 15 minutes.Media is being taken out from autoclave and pour it into the petridish.

PROCEDURE FOR PREPARATION OF MEDIA

A suspension of Microsporum fulvum and Trichophyton rubrum was added to the above sterile sabouraud media at 45oC and the mixture was transferred to sterile petri dishes and allowed to solidify.

Sterile 5.0 mm diameter blank discs were impregnated with test (extracts), standard (Miconazole) and control substances and were placed in petridishes containing a suitable agar medium seeded with the test organism (Trichophyton rubrum & Microsporum fulvum) using sterile loop.

The plates were made to stand for one hr at room temperature as a period of pre incubation diffusion to minimize the effect of variation in time between the applications of the different solutions.

Then the plates were incubated at 37oC for 18 hrs and observed for antifungal activity.

The diameters of the zones of inhibition were measured for the plates in which the zone of inhibition was observed.

Cultures should be examined at least weekly for fungal growth and should be held for 4 – 6 weeks before being reported as negative.

DISC DIFFUSION METHOD

The antifungal assay on acetone & benzene extracts of leucas lavandulifolia against pathogens (Trichophyton rubrum & Microsporum fulvum).

ANTIFUNGAL ASSAY

 Sl no

Fungal

Strain

Test Standard Control  Concentration

(mg/ml)Concentratio

n(mg/ml)

 

1 Trichophyton

rubrum

AE BE C1 -  1 1 1 -  

Zone of inhibition

(mm)

Zone of inhibition

(mm)

Zone of inhibition

(mm)  

11.8 14.7 17.5 - -  

 

2 Microsporum

fulvum

Concentration(mg/ml)

Concentration

(mg/ml)

Concentration(mg/ml)

AE BE C1 CONTROL1 1 1 -

Zone of inhibition

(mm)

Zone of inhibition

(mm)

Zone of inhibition

(mm)12.4 13.2 15.3 - -

OBSERVATIONS

Phytochemical tests The chemical tests of Leucas Lavandulifolia extracts

reveal the presence of alkaloids, carbohydrates, glycosides, saponins, phytosterols, phenols, tannins, flavanoids, proteins & amino acids.

 Results Anti-fungal assay of Leucas lavandulifoliaTest sample

RESULTS & DISCUSSIONS

PATHOGENS BENZENE EXTRACT

ACETONE EXTRACT

MICROSPORUM FULVUM

13.2 12.4

TRICOPHYTONRUBRUM

14.7HIGH

11.8LOW

PATHOGENS STANDARD SAMPLEMICROSPORUM

FULVUM15.3

TRICOPHYTONRUBRUM

17.5

Standard sample

Control sample The control doesn’t show any antifungal zone against any of the pathogens used.

THANK YOU