Natural Product Radiance

6
Research Paper Natural Product Radiance, Vol. 7(1), 2008, pp.6-9
Intro ducti o n
Cassia alata Li nn. , belonging
to family Caesalpiniaceae is a Pantr opical,
o r n a m e n t a l s h r u b , d i s t r i b u t e d fr o m
t r o p i c a l Am e r i c a t o I n d i a
1 , 2
. I t i s
commonly known as Ringwor m Senna.
The leaf extr acts of the plant have been
r epor ted to possess medicinal pr oper ties
and used against r ingwor m, scabies, ulcer s
and other skin diseases such as pr ur itis,
eczema and itching
3, 4
. Many r epor ts ar e
also available on antimicr obial activity of
its leaves
5-10
. The aim of the pr esent wor k
was to evaluate the antifungal activity of
its cr ude flower extr act on thr ee var ied
gr o u p s o f fu n gi n a m e l y a fl a t o xi n
pr oducing fungi ( Aspergillus flavus and
A. parasiticus) , plant pathogenic fungi
( Fusarium oxysporum a n d
Helminthosporium oryzae) a n d
h u m a n p a t h o ge n i c fu n gi ( Candida
albicans a n d Microsporum
audouinni) .
Mate ri al s and Me tho ds
Plant Materials: The flower s of C. alata
we r e c o lle c te d fr o m th e Co lle ge gr e e n
house, washed initially with 2% aqueous
NaCl solution followed by ster ile distilled
water, air dr ied under ster ile conditions
in s h a d e , p o wd e r e d a n d u s e d fo r th e
exper imental wor k.
Fungi s trai ns : The toxigenic str ains of
aflatoxin pr oducing fungi, Aspergillus
flavus ( NCBT 1 0 1 ) , A. parasiticus
( NCBT 1 2 8 ) ; p l a n t p a t h o ge n i c fu n gi
Fusarium oxysporum ( NCBT 1 5 6 ) ,
Helminthosporium oryzae ( NCBT
1 6 5 ) ; a n d h u m a n p a t h o ge n i c fu n gi ,
Candida albicans ( NCBT 1 4 0 ) a n d
Microsporum audouinni ( NCBT 173)
maintained at the Depar tment of Botany,
Na t i o n a l Co l l e ge we r e u s e d i n t h e
exper iment.
Expe ri me ntal pro ce dure : 100 mg of
dr y flower powder mixed with 20ml of
Sabour aud Dextr ose Agar ( SDA) medium
( HI Media M063) constitute 5 mg/ ml,
2 0 0 m g o f d r y fl o we r p o wd e r fo r
10 mg/ ml and 300mg for 15mg/ ml. The
contr ol ( a) contained only 20ml of SDA
medium, contr ol ( b) contained 100 mg
of bavistin and, contr ol ( b1) with 100mg
of gr iseofulvin fungicides added to 20ml
o f SDA me d iu m with ( 5 mg/ ml) . Ea c h
I n vitro antifungal activity of Cassia alata Linn. flower extract
M N Abubacke r
1
*, R Ramanathan
1
and T Se nthi l Kumar
2
1
PG and Resear ch Depar tment of Botany, National College,
Tir uchir appalli-620 001, Tamil Nadu, India
2
Depar tment of Industr y Univer sity Collabor ation,
Bhar athidasan Univer sity, Tir uchir appalli-620 024
*Cor r espondent author, E-mail: abubacker _ nct@yahoo.com
Phone: 0431-2482995, 2481997
Received 12 January 2007; Accepted 12 July 2007
Abstract
The aqueous flower extr act of Cassia alata Li nn. ( Family-Caesalpiniaceae) was
investigated for antifungal activity by agar diffusion method against thr ee distinct gr oups of fungi,
viz. aflatoxin pr oducing fungi, Aspergillus flavus ( NCBT 101) and A. parasiticus ( NCBT
128) ; plant pathogenic fungi, Fusarium oxysporum ( NCBT 156 ) and Helminthosporium
oryzae ( NCBT 1 6 5 ) ; an d h u man p ath o gen ic fu n gi Candida albicans ( NCBT 1 4 0 ) a n d
Microsporum audouinni ( NCBT 173) . Total inhibition ( 100%) of gr owth was seen at 10 and
15 mg/ ml concentr ations for aflatoxin pr oducing fungal str ains. Wher eas for plant and human
p a th o ge n ic fu n gi to ta l in h ib itio n wa s a t 1 5 mg/ ml c o n c e n tr a tio n . Th e min imu m in h ib ito r y
concentr ation ( MIC) values of the extr act var ied fr om 5.75 to 8.00mg/ ml for these fungi. Thus,
aqueous flower extr act of C. alata can be used as a potential antifungal agent for these thr ee
var ied gr oups of fungi.
Ke ywo rds : Antifungal activity, Cassia alata, Ringwor m Senna, Aflatoxin, Plant pathogenic
fungi, Human pathogenic fungi.
IPC co de ; Int. cl .
8
A61K 36/ 00, A61K 36/ 482, A61P 31/ 10
Twig of Cassia alata
Vol 7(1) J anuary-February 2008
7
Research Paper
petr idish ( 9cm) containing the medium
alone ( contr ol a) , medium with bavistin
( c o n tr o l b ) , me d iu m with gr ise o fu lvin
( b 1 ) , a n d fl o we r e xt r a c t mi xe d wi t h
m e d i u m wa s i n o c u l a t e d wi t h 0 . 5 m l
spor e suspension pr epar ed fr om 10 days
old cultur e and incubated for 6 days at
2 8

2
o
C t e m p e r a t u r e a n d k e p t i n
d a r k . Th r e e r e p lic a te s we r e p r e p a r e d
and inoculated for each tr eatment.
De t e r mi n a t i o n o f t h e Mi n i mu m
Inhibito ry Co nce ntratio n ( MIC)
MI C wa s d e t e r m i n e d b y t h e
liquid dilution method
11
. Dilution ser ies
wer e set up with 0.25 to 15.0mg/ ml of
Sa b o u r a n d gl u c o s e b r o t h m e d i u m .
To e a c h tu b e , 0 . 1 ml o f s ta n d a r d ize d
s u s p e n s io n o f fu n ga l s p o r e s ( 4 1 0
6
spor es/ ml) wer e added and included at
2 7 1
o
C fo r 2 4 h o u r s . Th e l o we s t
co n cen tr atio n wh ich d id n o t sh o w an y
gr o wt h o f t h e t e s t e d fu n gi a ft e r
micr oscopic evaluation was deter mined
as MIC.
Re s ul ts and Di s cus s i o n
Th e a q u e o u s e xtr a c t o f d r ie d
powder of C. alata flower has shown
good antifungal pr oper ties for all the thr ee
gr o u p s o f fu n ga l s tr a in s te s te d in th is
wor k.
The gr owth of aflatoxin pr oducing
to xige n ic Aspergillus flavus ( NCBT
101) and A. parasiticus ( NCBT 128)
fu n gi we r e to ta lly in h ib ite d a t 1 0 a n d
15mg/ ml concentr ations ( Table 1) . The
total inhibition r ate for A. flavus can be
co mp ar ab le to co n tr o l ( b ) , a stan d ar d
a n tifu n ga l te s t with b a vis tin tr e a tme n t
( 5 m g/ m l ) . Wh e r e a s 7 5 p e r c e n t o f
inhibition is compar able with contr ol ( b)
and that of A. parasiticus ( Fig.1) .
Tabl e 1 : Anti fungal acti vi ty o f fl o we r e xtract o f Cassia alata at 2 8
o
2
o
C
Fungus Co ntro l Co ntro l Fl o we r e xtract co nce ntrati o n
( a) ( b) ( mg/ ml)
5 1 0 1 5
Aspergillus flavus + + + + - + - -
Aspergillus parasiticus + + + + + + + + -
Fusarium oxysporum + + + + - + + -
Helminthosporium oryzae + + + + Co ntro l + + + -
( b1 ) 5 mg/ ml
Candida albicans + + + + + + + + -
Microsporum audouinni + + + + + + + -
+ + + + = Nor mal gr owth ; + + = 50% gr owth inhibition; + = 75% gr owth inhibition; -= Total inhibition
Contr ol ( a) = Medium without flower extr act ; Contr ol ( b) = Medium with bavistin
Contr ol ( b1) = Medium with gr iseofulvin
Fi g. 1 : Anti fungal e ffe ct o f crude fl o we r e xtract o f Cassia alata
( A) Aspergillus flavus, ( B) Aspergillus parasiticus, ( C) Fusarium oxysporum,
( D) Helminthosporium oryzae, ( E) Candida albicans, ( F) Microsporum audouinni
[ ( a) Contr ol without flower extr act, ( b) Contr ol with bavistin 5mg/ ml, ( b1) Contr ol with gr iseofulvin
5mg/ ml, ( c) Flower extr act 5mg/ ml, ( d) Flower extr act 10mg/ ml, ( e) Flower extr act 15mg/ ml]
Natural Product Radiance
8
Fo r p l a n t p a t h o ge n i c fu n gi
Fusarium oxysporum ( NCBT 1 5 6 )
a n d Helminthosporium oryzae
( NCBT 1 6 5 ) a 1 0 0 p e r c e n t t o t a l
i n h i b i t i o n o f gr o wt h wa s n o t i c e d a t
1 5 m g/ m l c o n c e n t r a t i o n o f a q u e o u s
extr act ( Table 1) . This total inhibition is
c o m p a r a b l e t o c o n t r o l ( b ) wi t h
b a vi s t i n t r e a t m e n t ( 5 m g/ m l ) fo r
b o t h F.

oxysporum a n d H.

oryzae
( Fig. 1) .
Th e h u ma n p a th o ge n ic fu n ga l
str ains, Candida albicans ( NCBT 140)
and Microsporum audouinni ( NCBT
173) wer e subjected to a 100 per cent of
i n h i b i t i o n o f gr o wt h a t 1 5 m g/ m l
c o n c e n t r a t i o n o f e xt r a c t ( Ta b l e 1 ) .
Wher eas 75 per cent of inhibition can be
seen for 10 mg/ ml concentr ation of extr act
and this is compar able with the contr ol
( b1) of gr iseofulvin tr eatment ( Fig.1) .
Th e stu d y r evealed th at a to tal
inhibition of gr owth of all the thr ee gr oups
of fungi tested for this wor k fall at two
levels of concentr ation of aqueous extr act
i. e. 1 0 an d 1 5 mg/ ml. Tr ad itio n ally leaf
extr act of this plant has been used to tr eat
fungal infections
12
. The r esults suppor t the
tr a d itio n a l u sa ge o f its flo we r s a ga in st
va r ie d gr o u p s o f fu n gi. Ea r lie r r e p o r t
2
suggested that C. alata leaves at 500mg/
m l c o n c e n t r a t i o n o f e t h a n o l s h o we d
antifungal activities against Aspergillus,
Penicillium, Microsporum a n d
Trichophyton sp p . Th e p r esen t wo r k
shows that lower concentr ation ( 15mg/
ml) o f aq u eo u s flo wer extr act is mo r e
e ffe c t i ve i n c o n t r o l l i n g t h e a fl a t o xi n
pr oducing, plant and human pathogenic
fungi. Accor ding to the ear lier wor k
2
the
leaf extr act h as n o an tifu n gal activities
a ga i n s t Candida albicans a n d
Cryptococcus neoformans, b u t th e
p r e s e n t wo r k p r o ve d t h a t t h e fl o we r
aq u eo u s extr act at 1 5 mg/ ml co n c. h as
go o d a n t i fu n ga l a c t i vi t i e s a ga i n s t C.
albicans.
Methanol extr acts of flower s at
4mg/ ml has been r epor ted to inhibit many
typ e s o f b a c te r ia , b u t n o t mo u ld s lik e
Aspergillus niger, Candida albicans
and Trichophyton mentagrophytes
9
.
An t h r a q u i n o n e s , c h r ys o p h e n o l a n d
flavonoid glycosides have been isolated
fr om C. alata and C. italica of which
the compound anthr aquinones have been
k n o wn t o p o s s e s s a n t i m i c r o b i a l
pr oper ties
13-15
.

C. alata bar k extr act is
a l s o r e p o r t e d t o p o s s e s s a n t i fu n ga l
a c t i vi t i e s e s p e c i a l l y t o wa r d s C.
albicans
10
. Th e p r esen t r ep o r t is also
evidence that the aqueous flower extr act
h a s a n t i fu n ga l a c t i vi t y a ga i n s t C.
albicans
10
.
The MIC values of the aqueous
extr act of flower var ied fr om 5.75mg/ ml
to 8.00 mg/ ml for the fungi tested. The
MIC value of F.

oxysporum ( NCBT 156) ,
A. parasiticus ( NCBT 128) , H. oryzae
( NCBT 165) , A. flavus ( NCBT 101) , M.
audouinni ( NCBT 173) wer e 5.75, 6.25,
6.50, 7.25 and 8.00 mg/ ml, r espectively.
Fu r th er in vestigatio n was p er fo r med to
demonstr ate the action of the extr act on
th ese fu n gi at d iffer en t co n cen tr atio n s.
The gr owth of these fungi cor r espondingly
decr eased with incr easing concentr ations
o f t h e e xt r a c t a n d t h e gr o wt h
wa s c o mp le te ly in h ib ite d a t th e ir MIC
va l u e s . Th e r e d u c t i o n o f gr o wt h wa s
possibly due to the inter fer ence by active
p r in c ip le s o f th e flo we r e xtr a c t. Su c h
inter fer ence may be at the biosynthetic
level. Ther efor e, the MIC deter mination
is impor tant in giving a guideline to the
c h o ic e o f a n a p p r o p r ia te a n d e ffe c tive
concentr ation of a antifungal ther apeutic
substance.
Co ncl us i o n
The aqueous flower extract of
C. alata is a s ign ific a n t in h ib ito r o f
gr owth of aflatoxin pr oducing, plant and
human pathogenic fungi tested dur ing this
investigation. Ther efor e, the identification
of this potential her b as antifungal agent
will help in r eplacing some commer cial
antifungal dr ugs.
Acknowl e dge me nts
Th e a u t h o r s t h a n k DST- FI ST,
Go ve r n m e n t o f I n d i a , Ne w De l h i fo r
pr oviding the infr astr uctur e facilities for
t h e De p a r t m e n t o f Bo t a n y, Na t i o n a l
College, Tir uchir apalli. The author s also
thank Sr i. K. Santhanam, Patr on Secr etar y,
Shr i K. Ragunathan, Secr etar y and Dr. K.
Anbar asu, Pr incipal, National College for
their encour agement.
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Research Paper