ANTIMICROBIAL ACTIVITY FROM THE

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ANTIMICROBIAL ACTIVITY FROM THE SEEDS OF
PSORALEA CORYLIFOLIA   Dissertation submitted to
the UNIVERSITY OF MADRAS   In the partial
fulfillment of the requirements For the Degree of
  POST GRADUATE DIPLOMA IN HERBAL
BIOTECHNOLOGY   By  DR. MD. OBIEDULLAH BAIG Reg.
No. 43830408     DEPARTMENT OF BIOTECHNOLOGY UNIVE
RSITY OF MADRAS, GUINDY CAMPUS CHENNAI 600
025  APRIL - 2005
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ACKNOWLEDGEMENT
I wish to express my sincere gratitude and
indebtedness to Dr. Shyamala , Reader, Department
of University of Madras for the help and expert
guidance in completing this project. I am
extremely grateful to Dr. P. ramasamy, Professor
and Head-In-Charge, Department of Biotechnology,
University of Madras for giving consent to carry
out this work in the department. I express my
sincere thanks to Ms. K. Uma Maheshwari,
Lecturer, Department of Biotechnology, University
of Madras for her valuable guidance and
suggestion rendered during the course of my work.
  I express my sincere thanks to the Research
Scholars of the Department of Biotechnology,
University of Madras for their help during the
project work.   I thank all my family and
friends for enabling me to complete this study
successfully.  (Md. Obiedullah Baig)
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INTRODUCTION
All over the world, plants have been one of the
most important sources of medicine since the
beginning of the human civilization. Tropical
forests touch the daily lives of every one on
earth through products and ecological services
they provide. The active ingredients for 25 of
the world's prescription drugs are substances
derived from plants, most of which grow in
tropical rain forests (the medicinal properties
of plants are associated with their chemical
constituents). About 80 of the world's
population relies on plant extracts for
medicines. Atleast 40 of all medicines, drugs
and pharmaceuticals worth 100 billion per year
owe their existence to the genetic resources of
wild plants, mostly from tropical forest of less
developed countries. About 70 of the 3,000
plants from tropical forests were identified by
the National Cancer Institute (NCI) as sources of
cancer fighting chemicals (Tyler Miller, JR
1996).   The herbal medicine represents
probably the first and certainly the oldest
system of human health care on the basis of
knowledge obtained through Rigveda,
Atharvanaveda, Charaka Sumheta (990 BC) and
Susruta Samheta (500 BC) for over 500 years.
Many ayurvedic pharmacopoeias have been compiled.
The chinese system of medicine is also equally
important, efficient and ancient. The unani
(Graceo Arabic) and Tibetan systems date back
to about 3000 years also stressed the importance
of herbal products for among sickness.   Herbal
medicine is still the mainstay of about 75-80 of
the world population, mainly in the developing
countries for primary health care. The use of
traditional medicine and medicinal plants in most
developing countries, as a normative basis for
the maintenance of good health has been widely
observed (UNESCO, 1996). The practice of
traditional medicine is wide spread in China,
India, Japan, Pakistan, Sri lank and Thailand.
In china 40 of the medicine is coming from
traditional medicine (Lemma, 1991).
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REVIEW OF LITERATURE
Psoralea corylifolia

• Name Babchi
• Biological Name Psoralea corylifolia
• Family Fabaceae
• Other Names Common and popular names of
  (Psoralea corylifolia) around the world
• Languages/regions/countries Names
• 1. Arabic Loelab el abid , Mahalep
• 2. Bengal Barachi, Bavachi, Hakuch,
  Latakasturi
• 3. Kanarese Bavanchi
• 4. Srilanka Ravoli
• 5. Chineses Ku Tzu, Pu Ku Chih
• 6. Gujarati Babchi, Bavacha, Bawachi
• 7. Hindi Babachi, Babchi, Bavanchi, Bukchi
• 8. Marathi Bavachya, Babchi, Bavachi
• 9. Persian Waghchi
• Sanskrit Alindavi,Asitatvacha,Avalguja,Bakuchi,Ch
  andrlekha,Chandraprabha, Chandraraji,Kalameshi,
  Kambojii,Kantaka,Krishna,Krishnaphala,sita,
  Sitavari,Samaraji,Suprabha,
• Sinhalese Bodi
• Tamil Karpokarishi, karpurvari,

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The Plant Psoralea corylifolia
An erect Herbaceous annual, 60-120cm high with
grooved and gland-dotted stems and branches
leaves simple, broad elliptic, rounded and
mucronate at the apex, covered with white hairs
on both surfaces, with numerous black dots, main
nerves 5, springing from the base flowers blue,
axillary, 10-30 flowered racemes fruits pods,
5mm long, subglobose, slightly compressed,
closely pitted, black, beaked without hairs,
seeds oblong, flattened, dark brown with an
agreeable aromatic odour and taste.
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Parts Used Seeds Constituents The chief
active principle of the seeds is an essential
oil and a fixed oil, a resin, and traces of a
substance of alkaloidal nature. Medicinal Uses
Anthelmintic Antibacterial Aphrodisiac
Astringent Cardiac Cytotoxic Deobstruent
Diaphoretic Diuretic Infertility Kidney
Odontalgic Skin Stimulant Stomachic Tonic.
Psoralia corylifolia is valued in Chinese herbal
medicine as a tonic remedy and is used to improve
general vitality. Modern research has shown that
it is also of value in the treatment of skin
disorders, including vitiligo. Some caution
should be employed when applying the herb
externally, however, since it can sensitise the
skin and cause an allergic reaction to sunlight.
Pharmacological action Antipleutritic
(decreases itching contains natural
antihistamine) antibacterial anthelmintic
laxative, diuretic seeds, oil has strong effect
against streptococci Alcoholic and aqueous
extracts of seeds antibacterial petroleum ether
extract anthelmentic against earth worms. In
dogs it stimulates intestinal smooth muscle
isopsoralene is tranquillosedative,
anticonvulsant, central muscle relaxant in rodent
model tests. Bavachinine shows anti-inflammatory
action in rat studies oleoresin extract of seeds
effective on leukoderma Antimicrobial
activity Antimicrobial activities of Psoralea
corylifolia seeds extracts were determined by the
well diffusion technique of Bauer-Kirby
(well-method) by measuring the zone of inhibition
(mm) against the bacterial species namely
Escherichia coli, Staphylococcus aureus, Proteus
mirabilis, Pseudomonas aeruginosa, Enterococcus
faecium. The extract showed no inhibition
against Shigella dysenteriae, Bacillus cereus,
Salmonella typhi, Actinomyces viscosus, Vibrio
cholerae.
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AIMS OF STUDY
The aim of the study was to determine the
antimicrobial activity of Psoralea corylifolia
seeds. The aqueous, chloroform ethanolic
extract of Psoralea corylifolia seeds were tested
against 10 different bacterial species by Well
diffusion Method.
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MATEIRALS AND METHOD

• Plant Material used
•  
• Psoralea corylifolia seeds were procured from the
  Agri-Horticulture society, Chennai, India.
•  
• Preparation of the extract
•  
• The seeds of the plant material were dried at
  31oC and were grounded into powder by using a
  electronic blendor. The Ethanolic, Chloroform
  and Aqueous extract of the seeds were prepared.
  50 gm of seeds was extracted with 350 ml of
  corresponding solvents in a soxhlet apparatus at
  room temperature for about 8 hours. The extract
  was filtered through Whatman no-1 filter paper
  and the residue was dried and used for the study.
  
•  
• Media used
•  
• Nutrients agar was prepared and sterilized by
  autoclaving at 121oC for 15 minutes.
• Sterile swabs, petridishes, microtips,
  pipettes, 0.85 saline, textube, etc.
• Bacterial species tested
•  
• The extracts were tested against Escherichia
  coli, Staphylococcus aureus, Proteus mirabilis,
  Pseudomonas aeruginosa, Enterococcus faecium,
  Shigella dysenteriae, Bacillus cereus, Salmonella
  typhi, Actinomyces viscosus and Vibrio cholerae.
  
•  
• Culture Media
•  

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Nutrient Agar Medium Peptone - 5
gm Yeast Extract - 2 gm Beef Extract -
1 gm Sodium Chloride - 5 gm Agar - 15
gm Distilled Water - 1000 ml pH - 7
0.2   Methods The antibacterial activity of
the ethanolic, chloroform and aqueous extract of
the seeds were determined by well diffusion
method.   The sterile nutrient agar was poured
on to the Petridishes and the media was allowed
to solidify. The bacterial colonies were
suspended in 0.85 saline and turbidity was
adjusted to match 0.5 Mc Farland standard. The
suspension of the bacteria were spread over the
agar having a sterile cotton swab. A well of
5mm diameter was cut on the agar with sterile
bores. The extracts of concentration of 20mg,
40mg, 80mg were added to the wells using
micropipettes. The plates were incubated at 37oc
for 24 hrs and examined. The diameter of the
zone of inhibition (mm) was measured for each of
the extract against each bacterial species.
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METHODS
Methods The antibacterial activity of the
ethanolic, chloroform and aqueous extract of the
seeds were determined by well diffusion method.
  The sterile nutrient agar was poured on to the
Petridishes and the media was allowed to
solidify. The bacterial colonies were suspended
in 0.85 saline and turbidity was adjusted to
match 0.5 Mc Farland standard. The suspension of
the bacteria were spread over the agar having a
sterile cotton swab. A well of 5mm diameter
was cut on the agar with sterile bores. The
extracts of concentration of 20mg, 40mg, 80mg
were added to the wells using micropipettes. The
plates were incubated at 37oc for 24 hrs and
examined. The diameter of the zone of inhibition
(mm) was measured for each of the extract against
each bacterial species.
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RESULTS
The antibacterial activity of the aqueous,
chloroform and ethanolic extract of Psoralea
corylifolia seeds against 10 different bacterial
species were tested.   Among the 10 bacterial
species tested Escherichia coli, Staphylococcus
aureus, Proteus mirabilis, Pseudomonas aeruginosa
and Enterococcus faecium were inhibited by all
the extracts of Psoralea corylifolia seeds. The
extracts did not show any activity against
Shigella dysenteriae, Bacillus cereus, Salmonella
typhi, Actinomyces viscosus and Vibrio cholerae.
  Of the three extracts tested for the
antibacterial activity, the aqueous extract of
the seeds showed more activity. The zone of
inhibition of Escherichia coli were 10mm, 14mm
and 15mm in diameter for 20mg, 40mg 80mg / ml
concentration respectively. Zone of inhibition
of the aqueous extract of seeds against
Escherichia coli, Staphylococcus aureus, Proteus
mirabilis, Pseudomonas aeruginosa and
Enterococcus faecium at 80mg / ml concentration
were 15mm, 18mm, 15mm, 13mm and 13mm diameter
respectively.   The zone of inhibition of the
chloroform and ethanolic extract against
Escherichia coli were 14mm and 10mm at 40mg / ml.
The Staphylococcus aureus was inhibited at 20mg,
40mg and 80mg and the zone of inhibition were
measured as 12mm, 15mm and 18mm respectively.
  The antibacterial effect of ethanolic extract
was found to be less inhibitory against
Staphylococcus aureus. All the three extracts
tested against Proteus mirabilis were inhibitory
at 80mg with a zone of inhibition of 15mm. The
chloroform extract of seeds showed a zone of
inhibition of 19mm at 80mg concentration and 15mm
for the ethanolic extract of seeds against
Enterococcus faecium.   The chloroform extract
of seed showed more antibacterial activity
against Pseudomonas aeruginosa at 80mg / ml
concentration and the zone of inhibition was
measured as 19mm. The ethanolic and aqueous
extract of seeds were less inhibitory against
Pseudomonas aeruginosa with a zone diameter of
13mm at 80mg / ml concentration. The
antibacterial activity and zone of inhibition
(mm) of the aqueous, chloroform and ethanolic
extracts of seeds of Psoralea corylifolia against
Escherichia coli, Staphylococcus aureus, Proteus
mirabilis, Pseudomonas aeruginosa and
Enterococcus faecium are shown in fig. 1, 2, 3.
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Antibacterial activity of the seed extracts of
Psoralea corylifolia
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Fig 2 Antibacterial effect of ethanol extracts
of seeds Well diffusion Method
Fig 1 Antibacterial effect of aquaous extracts
of seeds Well diffusion Method
Fig 3 Anti Bacterial effect of chloroform
extracts of seeds Well diffusion Method
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DISCUSSION
Plants have been one of the most important
sources of medicines. The screening and
isolation of bioactive compounds from plants is
increasing, as these compounds are much safer and
less toxic than the chemically synthesized drugs.
The increasing trends in emergence of resistance
among infectious agents, treatment failures and
recurrences of infection had led to the search of
biologically active compounds or products from
plants. The antibacterial activity of the seeds
of Psoralea corylifolia has already been
documented. The extract of seeds has shown
antibacterial activity against some of the oral
bacterial species tested. (Harumi Katsura et al
2001) Hence the study was done to evaluate
the antibacterial effect of seeds of Psoralea
corylifolia against some of the bacterial
pathogens. The aqueous, chloroform and ethanolic
extract of seeds of Psoralea corylifolia were
tested for their activity against 10 different
species. The extract showed no activity against
Shigella dysenteriae, Bacillus cereus, Salmonella
typhi, Actinomyces viscosus and Vibrio cholerae
where as Escherichia coli, Staphylococcus aureus,
Proteus mirabilis, Pseudomonas aeruginosa and
Enterococcus faecium were inhibited. The
aqueous extract of seeds showed more inhibition
than the chloroform and ethanolic extracts
against all the species of bacteria that were
inhibited. Antibacterial effect of aqueous
chloroform extract of seeds were higher than the
ethanolic extract against Staphylococcus
aureus. The chloroform extract showed more
inhibition against Pseudomonas aeruginosa, when
compared to the inhibitory action of the
ethanolic and aqueous extract of seeds. The
ethanolic extract of seed was more active against
Enterococcus faecium. The antimicrobial effect
of Psoralea corylifolia seeds may be due to the
presence of secondary metabolites, one of the
compound, Bakuchiol which is a alkaloid was found
to possess antibacterial effect on oral
bacterias. Hence further analysis of the
compounds and their testing against
micro-organisms would help to develop a new
antimicrobial agent that may be of great
importance.