Publication Ref No.

: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-10/DEC/017 ISSN 0974 – 9446

ANTI-INFLAMMATORY AND ANTIMICROBIAL ACTIVITY OF HEXANE EXTRACT OF SEED OF

PSORALEA CORYLIFOLIA LINN.

Bina Gidwani *1,

R. N. Alaspure 2, Dr. N.J. Duragkar2
1
Columbia Institute of Pharmacy, CSVTU,
Near Tekari, Raipur (C.G.), 492010, India,
2
.Sharad Pawar College of Pharmacy, RTM,
BIna Gidwani Near Wanadongri, Nagpur (Maharashtra) India
Email:beenagidwani@gmail.com

ABSTRACT

The main objective of this study is to determine anti-inflammatory and antimicrobial activity of
hexane extract of seeds of of Psoralea corylifolia Linn. Anti-inflammatory activity of Psoralea corylifolia
Linn. was evaluated by carrageenan induced rat paw oedema assay. The seed of plant was extracted
in soxhlet apparatus using hexane as solvent. Nine healthy swiss albino rats of either sex were divided
into 3 groups, each consisting of 3 rats. The evaluation was done by Hind paw method and compared
with standard and control groups. Further, the antimicrobial activity of Psoralea corylifolia Linn was
studied by Disc diffusion method using different gram positive and gram negative bacterial and fungal
strains, which showed that it has prominent activity compared with the standard used. The hexane
extract of Psoralea Corylifolia showed significant anti- inflammatory activity (44%) when compared with
the standard (55%). For antimicrobial activity, zone of inhibition were found to be in concentration of
5µg / ml for bacterial strains and 4µg / ml for fungal strains.

Key Words: Psoralea corylifolia, Anti – inflammatory activity, Anti – microbial activity, bawchi seeds,
carrageenan induced method, disc diffusion method.
.

INTRODUCTION 1. Side effects of modern drugs in many

Now-a-day’s interest in natural products as
potent therapeutic agent has increased
tremendously, the modern clinicians are now
inclined towards the use of herbal medicine1, 2, 3
The reasons for that are:
cases
2. Efficiency of a number of phyto
pharmaceuticals and herbal drugs which
are often enhanced by the presence of
minor constituents having synergistic
action 4

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Herbal medicine use as Anti microbial:
Infectious diseases account for a high
proportion of health problems in the developing
countries like India. Microorganisms have
developed resistance to many antibiotics and
this has created immense clinical problem in
treatment of infectious diseases. The resistance
of the organisms increased due to
indiscriminate use of commercial antimicrobial
drugs commonly used for the treatment of
infectious diseases. This situation forced the
scientists to search for new anti microbial
substances from various sources including
medicinal plants.5
According to WHO, more than 80% of
the world’s population relies on traditional
medicine for their primary health care needs.
Plants used for traditional medicine contain a
wide range of substances that can be used to
treat chronic as well as infectious diseases. The
medicinal value of plants lies in some chemical
substances that produce a definite physiological
action on the human body. The most important
of these bioactive compounds of plants are
alkaloids, flavonoids, tannins and phenolic
compounds.
Anti microbial activity of plants can be detected
by observing the growth response of various
microorganisms to those plant tissue or
extracts, which are placed in contact with them.
In order to detect anti microbial activity, three
conditions must be fulfilled.6
1. The plant extract must be brought in contact
with the cell wall of the microorganisms that
have been selected for test.
2. Conditions must be adjusted such that the
microorganisms are able to grow when no
antimicrobial agents are present.
3. There must be some means of judging the
growth, if any made by test organism.
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Psoralea corylifolia Linn commonly
known as babchi, belonging to family
Leguminoseae is a road side herbaceous weed,
found in Bengal, Bombay, through out Indian
plains. According to Ayurveda, there are 44
species of Psoralea out of which 4 are exotic
and this species is widely used in skin
disorders. The active constituents of seeds are
essential oil, alkaloids, tannins, saponins and
traces of lignins. Furanocoumarins are the
major constituents in seed coat i.e Psoralen and
Bakuchinin.4 Fixed oil, starch and proteins are in
kernel and amygdalin in palisade cells of the
seed coat. Other constituents are stigmasterol
in unsaponified matter and fatty acid
composition of fixed oil are palmitic, oleic,
linoleic and stearic acids. Roots of the plant are
useful in dental caries, leaves are good for
diarrhea, fruits are laxative, aphrodisiac and in
inflammatory diseases of the skin.7, 8,9,10,11
OBJECTIVE
Psoralea corylifolia Linn is a well known plant
widely used for skin disorders, the main
objective of this study was to determine the
anti–inflammatory and antimicrobial activity of
hexane extract of Psoralea corylifolia seeds and
further utilize it for different pharmacological
investigations.
EXPERIMENTAL WORK
Plant material
The entire plant Psoralea corylifolia Linn was
collected from the local region of Nagpur
district. The plant specimen was dried and its
herbarium sheet was prepared and
authenticated from the Department of Botany,
Rashtrasant Tukadoji Maharaj Nagpur
University, Nagpur. The authentication sheet no
9135. The seeds were collected from the plant
and dried before extraction.
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Preparation of the extract
For the investigation purpose, the seeds of the
plant were collected, air dried in shade, under
normal environmental condition and then
subjected to size reduction to get coarse
powder. Such powdered material was charged
into the soxhlet apparatus, and extraction was
carried out using n-hexane as solvent.
Evaluation of anti – inflammatory activity
12,13
Both in-vivo and in-vitro methods are available
for the evaluation of anti-inflammatory agents
but among them in-vivo method the
carrageenan induced rat paw oedema assay is
believed to be one of the most reliable and also
the most widely used. Carrageenan is a mixture
of polysaccharides composed of sulfated
galactose units and is derived from Irish Sea
moss, Chondrous crispus. The oedema, which
develops in rat paw after carrageenan injection,
is a biphasic event. The initial phase is
attributed to the release of histamine and
serotonin, the oedema maintained between the
1st and 2nd phase to kinin like substances and
the 2nd phase to prostaglandins like compound.
The carrageenan assay method is
advantageous because,
1. The oedema is specifically inhibited by anti-
inflammatory compounds.
2. Single oral dose of drugs at non-toxic levels
are effective.
3. Low variability.
4. Better reproducibility.
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5. Carrageenan itself is neither antigenic nor
causes any systemic effects.
Animals
Nine healthy albino rats of either sex weighing
100 – 150 g each were taken and divided into 3
groups each consisting of 3 rats. This study was
approved by Ethical Committee and the Reg.
No. is 536/02/CPCSEA The animal work was
done in Sharad Pawar College of Pharmacy,
Nagpur on 15 December/SPCP/2007
Procedure
Group I: The first group served as control
and received normal saline.
Group II: The second group received the test
extract of Psoralea corylifolia.
Group III: The third group served as standard
and received Diclofenac sodium.
The first group was given normal saline,
the second group served as test, which
received extract of Psoralea Corylifolia 25mg/kg
and 50mg/kg and the third group received
diclofenac sodium 10mg/kg and was used as
standard. Before administration of test drug, the
hind paw thickness of all rats was measured by
using digital vernier caliper. Soon after the
measurement the drugs were administered
orally. One hour later subcutaneous injection of
0.1 ml of 1% w/v carrageenan in water was
injected into the planter surface of both hind
paws of each rat. The paw thickness was
further evaluated at an interval of 90 min, 180
min, 270 min and 360 min. 14
The activity of drug was expressed as percent
inhibition of oedema.
Percent inhibition = 100 * (1 – Tt/Tc)
Where, Tt and Tc are the average
increase in paw thickness of drug treated and
control group respectively.
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Antimicrobial activity 15, 16, 17
The antimicrobial activity (antibacterial and
antifungal) was done by disc diffusion method.
The basic idea of diffusion assay is as follows:
A suspected antimicrobial compound or
treatment is presented within a reservoir
created on an inoculated plate of agar medium;
following diffusion of the compound(s) through
the agar, “zone of inhibition” forms where
concentrations of the diffused molecules is
sufficient to inhibit microbial growth. Diffusion of
antimicrobial compounds from a reservoir over
time produces an outward gradient of
decreasing concentration of the compound.
Where concentration of the compound is
sufficient to inhibit the growth of the microbes,
the growth is blocked; resulting in the observed
zone, which extends outward from the reservoir
(with a corresponding decrease in
concentration) to the distance from the reservoir
at which the minimum concentration required for
inhibition exists.
2.4.1 Strains used
The Antimicrobial activity of Psoralea corylifolia
extract was studied against bacterial strains viz.
E. coli (Gram-negative), S. aureus (Gram-
positive) and B. substilis (Gram-positive). The
fungal strains used were C. albicans and A.
niger. For antibacterial activity tetracycline was
used as standard, while for antifungal activity
Griseofulvin was used as the standard.
2.4.2 Composition of Media
The medium should be such that it will promote
aster growth of microorganisms. (Refer Table
No. 02 & 03)
TABLE 2: NUTRIENT AGAR MEDIA (FOR
ANTIBACTERIAL ACTIVITY)
TABLE 3: POTATO DEXTROSE AGAR MEDIA
(FOR ANTIFUNGAL ACTIVITY)
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Procedure
Screening of antimicrobial activity was carried
out using the disc diffusion method. This
method depends on the diffusion of drug from
disc through the solidified agar layer of a
petridish to an extent such that growth of the
inoculated microorganism is prevented entirely
in a circular area “zone” around the cup
containing the solution of the compound under
test.One loopful of the stock culture was
inoculated at 10 ml of agar slant previously in
sterilized test tubes, and incubated at 37o for 24
h and 20o for 48 h respectively for bacteria and
fungi. About 3 ml of distilled water was added to
the test tube and a suspension of the culture
was obtained by shaking for few minutes. The
solutions were made by dissolving the test
compound in minimum amount of DMSO, and
volume was made with sterilized water to
produce a concentration of 100 µg/ml. All the
operations were carried out under aseptic
conditions. Respective sterile medium was
melted on water bath and kept at 45o in
constant temperature water bath. In each sterile
petridish 25 ml of molten medium was added
and 107 /ml of sub cultured organism under
study were inoculated. The culture and agar
medium were mixed and allowed to solidify.
Four disc of whatmann No.41 (4 mm diameter),
which were dipped in test samples were then
kept on the upper surface of the medium.
Solution was allowed to diffuse in the medium
for 2 h by keeping the petridish at room
temperature and then incubated for about 24 h
at 37o (for bacteria) and 48 h at 20o (for fungi).
RESULT
The anti-inflammatory activity was evaluated
and the percentage of inhibition of the extract
was compared with the standard. The
percentage inhibition of the test extract was
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found to be 44%, compared to standard which lead to development of new potent compounds
is 55%. The observations and results are shown with significant activity.
in Table 1. (Refer Table No. 01)

TABLE 1 STUDY OF ANTI – INFLAMMATORY REFERENCES:

ACTIVITY OF EXTRACT OF PSORALEA
CORYLIFOLIA 1. WHO General guidelines for
Methodologies on Research and Evaluation of
Figure 1 and 2 shows the graphical herbal medicines. 2000; 14: 237.
representation of comparative anti-inflammatory 2. Chaudhari AC, Dixit SK. Quality Control
activity. The in-vitro antimicrobial activity was and Standardization of Herbal Raw materials
carried out by Disc diffusion method using and Traditional remedies. Eastern
different bacterial and fungal strains, which Pharmacist.1995; 3: 23.
showed that the oil posses’ prominent activity 3. Patani A. Indian Herbal
compared with that of standard. Zone of Pharmacoepoeia. Revised new ed. Indian drug
inhibition were found to be prominent in manufacturer association publications. Mumbai;
concentration of 5µg / ml for bacterial strains 2002. p. 550.
and 4µg / ml for fungal strains (Table 4 and 5). 4. Lai PK, Roy J (June 2004). "Antimicrobial
Figure 3 and 4 shows the antibacterial and and chemopreventive properties of herbs and
antifungal activities of hexane extract of seeds spices". Curr. Med. Chem. 11 (11): 1451–60.
of Psoralea corylifolia with the standard. 5. Rao MR, Reddy IB. Antimicrobial activity
of some Indian medicinal plants. Indian J.
Microbiology. 2006; 46.
DISCUSSION 6. The Indian Pharmacopoeia. Govt. of
India, Ministry of Health and Family Welfare.
On the basis of the result, it is concluded that The Controller of Publication, 1996; A-53, 54,
the extract of Psoralea corylifolia Linn. 89.
Possesses significant anti-inflammatory activity. 7. Mukherjee PK. Quality Control of Herbal
Also, the plant showed prominent antimicrobial Drugs.1st ed. Business Horizon Pharmaceutical
activity when compared with the standard. Publishers; 2002; 600.
Further, pharmacological studies can be done 8. Rangari VD, Agrawal SR. Chemistry and
on the basis of the results obtained. This Pharmacology of PSORALEA CORYLIFOLIA
relevant information is helpful for further Indian Drugs.1992; 29: 662.
investigation. 9. Kirtikar KR, Basu BD. Indian Medicinal
Plants.2nd ed. Mahendra publication; 994.p.
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techniques such as preparative TLC, column Encyclopedia. 1st ed. Shri Satguru
chromatography etc, it would be possible to publication; 1998;. 76.
evaluate and study other activities of the plant. 11. Rajpal V. Standardization of Botanicals.
These isolated constituents in pure form would 2nd ed. Eastern Publisher; 2005; 284.
possess prominent anti inflammatory, analgesic 12. Costa-lotufo LV , Lucena DF, Netu MA,
and anti microbial activity, and can be used in Bezerra JNS, Viana GSB. Analgesic,
treatment of various skin disorders. This can Antiinflammatory and Central Depressor Effects

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of the Hydroalcoholic Extract and Fractions activity of the seeds of Argimonia eupatoria.
from Aeolanthus suaveolens. Biol.Pharm. Fitoteripia. 2003; 73: 133.
Bull.2007; 27:821. 16. Khan MR, Khihana M, Omoloso AD.
13. Kulkarni SK. In; Handbookss of Antimicrobial activity of Michelia champaca
Experimental Pharmacology, 2nd ed. Vallabh .Fitoteripia 2003; 73:744.
Prakashan, New Delhi ; 2005;.127. 17. Pistelli, Betroli AT, Lepori DH.
14. Kupeli E , Tosun A, Yesilada E. Antimicrobial and antifungal activity of crude
Assessment of anti-inflammatory and anti- extracts and isolated saponins from Astragalus
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Ethanopharmacol 2007; 113:333.
15. Copland A, Nahar AT, Tomlinson CTM.
Antibacterial and free radical scavenging

TABLES AND FIGURES:

TABLE 1 STUDY OF ANTI – INFLAMMATORY ACTIVITY OF EXTRACT OF PSORALEA

CORYLIFOLIA

Percent inhibition
Compound 0 hr 1 hr 3 hr 5 hr
1 hr 3 hr 5 hr
7.24 ± 7.34± 7.19 ± 6.92 ±
Control - - -
0.93 0.99 1.06 1.23
Psoralea corylifolia 6.93 ± 6.23 ± 5.59 ± 4.21 ± 28.6 38.8
11%
extract (25mg/kg) 0.56 0.96 1.05 0.47 % %
Psoralea corylifolia 7.83 ± 6.88 ± 5.37 ± 4.43 ± 31.6
16% 44%
extract (50mg/kg) 0.48 0.29 0.48 0.15 %
Standard (diclofenac 7.55 ± 6.15 ± 4.77 ± 3.54 ± 21.5
43% 55%
sodium 10mg/kg) 0.88 0.15 0.09 0.16 %

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TABLE 2: NUTRIENT AGAR MEDIA (FOR ANTIBACTERIAL ACTIVITY)

Sr.No INGREDIENT QUANTITY
1. Beef extract 10.0g
2. Peptone 10.0g
3. Sodium chloride 5.0g
4. Agar 12.0g
5. water Upto 1000ml
TABLE 3: POTATO DEXTROSE AGAR MEDIA (FOR ANTIFUNGAL ACTIVITY)

Sr.No INGREDIENT QUANTITY

1. Potato infusion 0.2g

2. Dextrose 20g

3. Agar 15g

4. Water Upto 1000ml

TABLE 4: ANTIBACTERIAL ACTIVITY

Zone of Inhibition in mm
Microorganism Psoralea extract Tetracycline
1µg/ ml 2µg/ ml 3µg/ ml 4µg/ ml 5µg/ ml (5µg / ml)
E. coli 6 8 11 13 15 17
B. subtilis 5 7 9 11 14 21
S. aureus 7 9 13 15 19 23

TABLE 5: ANTIFUNGAL ACTIVITY

Microorganism Zone of Inhibition in mm

Psoralea extract Griseofulvin
1µg / ml 2µg / ml 3µg / ml 4µg / ml 5µg / ml (5µg / ml)
C. albicans 9 11 16 20 27 25
A. niger 7 9 11 14 19 17

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Figure 1 and 2 shows the graphical representation of comparative anti-inflammatory activity..

Fig.1: Anti-inflammatory activity of Psoralea extract with standard Diclofenac sodium

10.0 Control
PCO 25mg
PCO 50mg
paw thickness

7.5 Std.

5.0

2.5

0.0
0 1 2 5
Hours

Fig.2: Percent inhibition of paw thickness of rat by Psoralea extract & Diclofenac sodium

P C O 25mg
60 P C O 50mg
S td
50
% Inhibition

40

30

20

10

0
1 2 5
H o ur s

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Fig. 3: Antibacterial activity of Psoralea extract and Tetracycline

Antibacterial activity
Zone of Inhibition

25
20 Psoralea oil
in mm

15 (5µg/ml)
10 Tetracycline
5 (5µg/ml)
0
1 2 3
Fig. 3: Antibacterial activity of Psoralea extract and Tetracycline
Bacterial strains

Fig. 4: Antifungal activity of Psoralea extract and Griseofulvin

Antifungal activity
Zone of Inhibition in

30
25
Psoralea oil
20
(4µg/ml)
mm

15
Griseofulvin
10
(5µg/ml)
5
0
1 2
Fig. 4: Antifungal activity of Psoralea extract and Griseofulvin
Fungal strains

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