EVALUATION OF ANTIFUNGAL POTENTIAL OF LEUCAS LAVANDULIFOLIA

1. 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)

2.  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.

4. 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.

5. 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.

6.  Observations
BE: Benzene extract; AE: Acetone extract; ‘+’: presence of phytochemical; ‘-’: absence of phytochemical.
PHYTOCHEMICAL
CONSTITUENTS
L. Lavandulifolia
BE AE
ALKALOIDS + +
CARBOHYDRATES - +
GLYCOSIDES + -
SAPONINS + +
PHYTOSTEROLS - +
PHENOLS + -
TANINS + +
FLAVANOIDS + +
PROTIENS - -
AMINO ACIDS - -
.

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

8. 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.

9.  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.

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

11. Sl
no
Fungal
Strain
Test Standard Control
Concentration
(mg/ml)
Concentration
(mg/ml)
1 Trichop
hyton
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 Microsp
orum
fulvum
Concentration
(mg/ml)
Concentratio
n
(mg/ml)
Concentration
(mg/ml)
AE BE C1 CONTROL
1 1 1 -
Zone of inhibition
(mm)
Zone of
inhibition
(mm)
Zone of inhibition
(mm)
12.4 13.2 15.3 - -

12. 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 lavandulifolia
Test sample
PATHOGENS BENZENE EXTRACT ACETONE EXTRACT
MICROSPORUM
FULVUM
13.2 12.4
TRICOPHYTON
RUBRUM
14.7
HIGH
11.8
LOW

13. PATHOGENS STANDARD SAMPLE
MICROSPORUM
FULVUM
15.3
TRICOPHYTON
RUBRUM
17.5
Control sample
The control doesn’t show any antifungal zone against any of the
pathogens used.