Original Article

Antibacterial activities and phytochemical analysis of

Cassia fistula (Linn.) leaf
Sujogya K. Panda1,2, L. P. Padhi2,
G. Mohanty1 Abstract
Departments of 1Biotechnology, and Cassia fistula Linn. which belongs to family Leguminosae is a medium-sized tree and its
2
Zoology, North Orissa University,
different parts are used in ayurvedic medicine as well as home remedies for common
Baripada, Orissa, India
ailments. Sequential extraction was carried out using solvents viz. petroleum ether,
J. Adv. Pharm. Tech. Res. chloroform, ethanol, methanol and water from leaf of the plant were investigated for
preliminary phytochemical and antibacterial property. Results of the study showed
that all the extracts had good inhibitory activity against Gram-positive test organism.
Although all five extracts showed promising antibacterial activity against test bacterial
species, yet maximum activity was observed in ethanol extract. The minimum inhibitory
concentration ranged in between 94 to 1 500 μg/ml. Evaluation of phytochemicals such
as alkaloids, flavonoids, carbohydrates, glycosides, protein and amino acids, saponins,
and triterpenoids revealed the presence of most of constituents in polar extracts
(ethanol, methanol, and aqueous) compared with nonpolar extracts (petroleum ether
and chloroform). Furthermore, the ethanol extract was subjected to TLC bioautography
and time-kill study against Staphylococcus epidermidis. All the findings exhibit that the
leaf extracts have broad-spectrum activity and suggest its possible use in treatment
of infectious diseases.

Key words: Cassia fistula, human pathogenic bacteria, minimum inhibitory concentration,
Similipal Biosphere Reserve, TLC bioautography

INTRODUCTION on hepatoprotective, [4] antifertility, [5] and antioxidant

properties of C. fistula.[6,7] Some studies have also been done
Cassia fistula Linn. (Leguminosae) is a very common plant on antimicrobial activity of C. fistula flower and seed along
and is widely known for its medicinal properties. In the with some other Indian medicinal plants.[8-16] These studies
Indian literature, this plant has been described to be useful give diminutive information on the antimicrobial property
against skin diseases, liver troubles, tuberculous glands of the leaves of this plant.
and its use in the treatment of rheumatism, hematemesis,
pruritus, leucoderma, and diabetes.[1,2] Besides, it has been Reports of the preliminary screening of this plant collected
found to exhibit anti-inflammatory and hypoglycemic from Similipal Biosphere Reserve (SBR)[17] recorded of its
activity and widely used as a mild laxative suitable for antimicrobial activity against certain bacterial strains. The
children and pregnant women.[3] Several reports are present plant has diverse ethnomedicinal uses by the tribals of SBR.
The bark paste is applied externally on the bite area for 2 to
Address for correspondence
3 times in a day at regular interval for 3 days. Half teaspoon
Dr. Sujogya K. Panda,
Department of Biotechnology, North Orissa University, Baripada juice extract is taken orally thrice daily to cure jaundice.
- 757 003, Orissa, India. E-mail: sujogyapanda@gmail.com The leaf paste along with neem is applied externally over
all types of skin infections. Several reports are available on
Access this article online antimicrobial activities of C. fistula from bark, seed, flower,
Quick Response Code: and fruits, but that on leaves are scanty.
Website:
www.japtr.org In the present study, an a empt has been made to investigate
the antibacterial activity of different solvent extracts of leaf of
DOI: C. fistula obtained by sequential extraction method, against
10.4103/2231-4040.79814 Gram-positive and -negative bacteria. In addition to this,
TLC bioautography study and presence of phytochemical

62 Journal of Advanced Pharmaceutical Technology & Research | Jan-Mar 2011 | Vol 2 | Issue 1
 Panda, et al.: Antibacterial activity of Cassia fistula leaf

constituents in the respective extracts were also carried out. mean±SD of the inhibition zone was taken for evaluating
the antibacterial activity of the extracts.
MATERIALS AND METHODS
Determination of Minimum Inhibitory Concentration
Plant Material Although the results of the ACM cannot always be compared
Leaves of C. fistula were collected in the month of April, with the minimum inhibitory concentration (MIC) data,
2007 from SBR, Mayurbhanj, Orissa and their identity extracts which showed positive result were further evaluated
was confi rmed and the voucher specimen (NOU 25) for the determination of MIC. A broth microdilution
was deposited in the Department of Botany, North technique was adopted using 96-well microtiter plates and
Orissa University. The shed dried healthy leaves were tetrazolium salt, 2,3,5-triphenyltetrazolium chloride (TTC)
powdered separately using mechanical grinder and then was carried out to determine the MIC following the methods
were passed through sieve in order to maintain uniform with modification, as described by Eloff.[19] In the plate, A1 to
powder size. H1 were blank with MH broth only. A3 to H3 were having the
stock solution of the test extract(s) and A4 to H4 till A9 to H9
Preparation of Extracts were the wells in which the test extracts were serially diluted
Sequential extraction was carried out with the same powder using MH broth. Wells A12 to D12 were controls having 20 μl
using solvents of increasing polarity. About 250 g of dry of DMSO and E12 to H12 served as control over control. All
leaf powder were sequentially extracted using petroleum wells were dispensed with 100 μl of MH broth. 20 μl of the
ether, chloroform, ethanol, methanol, and aqueous solution herbal extract was transferred from stock test solution to
in Soxhlet apparatus. A er about forty siphons of each the first well, that is, from A4 to H4 containing 100 μl of MH
solvent extraction step, the materials were concentrated by broth. 20 μl of the MH broth containing herbal extract was
evaporation. The concentrated extracts were freeze dried to then transferred to the next well to create serial dilutions.
remove the solvent at -2oC till further use. The yield of each 100 μl of the 0.5 McFarland adjusted activated culture in MH
extract was calculated and stored for further use. broth was then added to all the wells except the blank. 5 μl
of 0.5% TTC was further added to all the dilutions, blank,
Bacteria and Growth Media control, and control over control. The final volume of all the
Escherichia coli (MTCC 1098), Escherichia coli O157:H7 wells was 205 μl. The microplate was sealed and incubated
(RMRC, Bhubaneswar, India), Salmonella typhimurium at 37°C at 130 rpm. 10 μl of the broth from each culture tube
(MTCC 3216), Shigella sonnei (NICED, Kolkata, India), exhibiting MIC and control tubes were taken aseptically
Bacillus subtilis (MTCC 7164), Bacillus licheniformis (MTCC and were plated on one-day old Muller-Hinton (MH) agar
7425), Staphylococcus aureus (MTCC 1144), and Staphylococcus plate as a point inoculum and allowed to dry for 10 minutes
epidermidis (MTCC 3615) were used as test microorganisms. under the laminar air hood. The microplate was sealed and
All these bacterial cultures were grown at 37 oC and incubated at 37oC at 130 rpm and observed for growth of
maintained at 4oC on nutrient agar slants (Hi-media Pvt. the microorganism. The lowest recorded MIC was further
Ltd., Mumbai, India). subjected to time kill kinetics using ethanol, methanol, and
aqueous extracts of C. fistula. 20 μl of overnight broth culture
Agar Cup Method of S. epidermidis was added to 180 μl of NB containing each
The agar cup method (ACM) was used to study the extract. The microtiter plate was incubated at 37°C. The
antibacterial activity of the extracts.[18] Briefly, cultures number of viable cells was determined a er 0, 1, 2, 4, 8, 12,
of the bacteria from culture plates were scooped using a 16, 24, and 48 hours of incubation. A control culture without
wire loop and separately mixed with normal saline and crude extract was incubated and assayed under the same
agitated with vortex mixer. A loop full was withdrawn condition.[20]
and uniformly distributed on the surface of the agar plate
by streaking using a sterile swab. Wells of approximately TLC Bioautography
6 mm in diameter and 2.5-mm deep were made on the TLC bioautography assay was performed by agar overlay
surface of the solid medium using a sterile borer. The plates bioautography technique. Plant extract samples (5 μl)
were turned upside down and the wells labeled with a were applied 2.5 cm from the base of the silica plate. A er
marker. The extracts were reconstituted by dissolving in drying, the plates were developed using solvent chloroform
dimethyl sulfoxide (DMSO). Each well was filled with test : methanol (8.2 : 1.8) and chloroform : hexane (5.4 : 6.6).
sample. Sterile DMSO was used as negative control, while Developed TLC plates were carefully dried for complete
gentamicin and ciprofloxacin were used as positive control. removal of solvents. Aliquot of 50 ml of molten MH agar
The plates were incubated at 37°C for 24 hours. A er 24 was prepared by adding 500 μl of bacterial inoculum (5 
hours, the plates were removed and zones of inhibition 105 CFU). Now, the inoculum containing agar was overlaid
measured with Himedia antibiotic scale and the results on dried TLC plate under aseptic condition in laminar
were tabulated. Extracts with zones of inhibition greater airflow. The plates were incubated at 37oC and examined
or equal to 8-mm diameter were regarded as positive. The for the zone of inhibition.

Journal of Advanced Pharmaceutical Technology & Research | Jan-Mar 2011 | Vol 2 | Issue 1 63
 Panda, et al.: Antibacterial activity of Cassia fistula leaf

RESULTS AND DISCUSSION test extracts. Several techniques have been reported for
the testing of antimicrobial activity of natural products
The amounts of crude material extracted per gram of including plant extracts at different time and stipulation.
powdered C. fistula leaf were 15.2, 24.5, 43.8, 68.2, and However, ACM is always employed for preliminary
158.6 mg, respectively, with chloroform, petroleum testing of antimicrobial activities of crude extracts, while
ether, methanol, distilled water, and ethanol solvents. advanced method like MIC is mostly used as secondary
Evaluation of phytochemicals such as alkaloids, flavonoids, confirmation method. In ACM, all test bacteria showed zone
carbohydrates, glycosides, protein and amino acids, of inhibition with petroleum ether, ethanol, methanol, and
saponins, and triterpenoids revealed the presence of most of aqueous extracts. Among Gram-positives, S. epidermidis,
the constituent in polar extracts such as ethanol, methanol, B. licheniformis, and B. subtilis were absolutely inhibited by
and aqueous extracts compared with nonpolar extracts all extracts. Ethanol and methanol extracts of the leaf were
(petroleum ether and chloroform). However, flavonoids, most active inhibiting agent against both Gram-positive
proteins and amino acids, tannins, and phenols were found and -negative bacteria. On the other hand, petroleum
to be universally occurring in all the extracts [Table 1]. ether extract had be er antibacterial activity against most
of the Gram-negative bacteria. Among Gram-negatives,
Table 2 lists the plant extracts and their zone of inhibition the highest zone of inhibition was recorded for ethanol
against the test bacterium. Chloroform extract did not show extract against E. coli O157:H7. From ACM, result was
any zone of inhibition against Gram-negatives, whereas obtained that there was no significant difference among
Gram-positive bacteria were completely inhibited by all the test bacteria with respect to plant extracts, while there
were marked differences between the activities of the
Table 1: Phytochemical screening of C. fistula plant extract and pure antibacterial drug (ciprofloxacin)
leaf [Figure 1]. Such significant differences are normally present
Name of the Qualitative test PT CH ET MT AQ when crude (unpurified) plant extracts are compared with
phytochemical pure drugs that are already in clinical use.[21] Also, the
Alkaloids Mayer’s reagent - - + + + ACM is not always dependable for accurate assessment
Dragendorff’s reagent - - - - - and comparison. This is because of the high degree of
Hager’s reagent - - + - + interference inherent in this method that arises from drug
Wagner's reagent - - - - - diffusion problems.[22]
Carbohydrates Molisch’s test - - - - -
Fehling’s test - - - + + The MIC results showed that all the extracts were able to
Benedict’s test - - - + + prevent the growth of all test organisms with selective
Tannin and With Ferric chloride + + + + + activities. The growth of inhibition of test bacteria range
phenolic With lead acetate + - + - + from 94 to 1 500 μg/ml (w/v), with the lowest MIC value
compound With gelatin solution - - - - - against S. epidermidis Table 3. The results of MBC showed
Glycoside Keller-Kiliani test - - + + - that in a concentration of 1 500 μg/ml (w/v), most of the
Legal test - - - - - organisms from both Gram-positive and-negative bacteria
Borntrager's test - - - - - were killed. These results clearly indicated that S. epidermidis
Protein and Biuret test - - + + + was the most sensitive organism, which is inhibited with
amino acids Ninhydrin test - - + + + MIC values lower than 100 μg/ml recorded in 50% of the
Xanthoprotein test - - - - - test cases. MIC values lower than 200 μg/ml were also
Millon’s test - - - - -
Gum and Molisch’s test - - - - -
mucilages
Flavonoids With NaOH + + + + +
With H2SO4 - + + + +
With Mg/HCl - - + + +
Saponins Honeycomb foam - - - - -
Foam test - - + + +
Steroids and Salkowski’s test - - - - -
sterol Libermann burchard - - - - -
Triterpenoids Thionyl chloride test + - + + -
Oils and fats With filter paper - - - - -
With alkaline KOH - - - - -
Vitamin C With Indophenols - - - - -
Sodium nitroprusside - - - - -
+: Presence; -: Absence Figure 1: Time kill kinetics against S. epidermidis

64 Journal of Advanced Pharmaceutical Technology & Research | Jan-Mar 2011 | Vol 2 | Issue 1
 Panda, et al.: Antibacterial activity of Cassia fistula leaf

Table 2: Antibacterial screening of different extracts of C. fistula leaf

Name of the strain Zone of inhibition in mm Ciprofloxacin
PE CH ET MT AQ
Gram-negative
Escherichia coli 13.0±0.5 13.6±0.5 12.6±0.5 12.6±0.5 11.6±0.5 19.6±2.0
Escherichia coli O157:H7 12.6±0.7 - 14.6±0.7 12.6±0.7 13.3±1.4 23.0±2.0
Salmonella typhimurium 12.6±0.5 - 13.6±0.5 13.3±0.5 12.6±0.5 24.6±1.5
Shigella sonnei 12.3±0.6 - 15.0±1.0 14.3±0.5 12.3±1.5 26.6±1.1
Gram-positive
Bacillus subtilis 14.6±0.7 12.3±0.7 14.6±0.7 12.6±0.7 13.6±0.7 16.6±1.5
Bacillus licheniformis 12.6±1.4 14.3±0.7 14.3±0.7 12.3±1.4 13.3±1.4 15.6±0.5
Staphylococcus aureus 10.6±0.5 10.6±0.5 13.6±0.5 13.3±0.5 12.6±0.5 29.0±2.0
Staphylococcus epidermidis 15.6±0.7 12.0±0.7 16.0±2.1 12.3±1.4 13.0±0.7 26.6±1.5
All values are mean zone of inhibition±SD (-) No zone of inhibition; Zone of inhibition including 6 mm borer; Extract concentration (30 mg/ml)

obtained with other extracts viz. ethanol, methanol, and

aqueous against B. subtilis, B. licheniformis, and S. epidermidis.
This activity could be considered very promising for two
reasons, first, the test bacteria were resistant to the first line
antibiotics and second, when compared with the reference
drugs’ (standard antibiotics) MICs (10 μg/ml). However, all
lowest MICs exhibited by extracts with MBC value eight
times of MIC in corresponding microorganisms highlighting
their interesting antimicrobial potency. From these results,
it can be observed that most of the tested samples exert
a killing effect on the test organisms, as MBC and MFC
values were recorded. However, when analyzing carefully
the MIC and MMC results, it can be noted that MMC/
MIC ratios lower than 4 was obtained with most of the
studied samples, suggesting that killing effects could be
expected.[23] The TLC results showed a total of 7 spots with
Rf values 0.09, 0.43, 0.40, 0.375, 0.55, 0.64, and 0.72. Ethanol,
methanol, and aqueous extracts had three similar Rf values
(0.09, 0.43, 0.40) than other extracts, which was possible
responsible for be er antibacterial activity. Ethanol extract Figure 2: TLC Bioautography with S. epidermidis
showed 5 distinct fractions with Rf value 0.72, 0.55, 0.43,
0.37, and 0.09. The same TLC plate of ethanol extract was compared with that of methanol and aqueous.
subjected against S. epidermidis to study bioautography and
result showed that the activity was limited to the fraction Kumar et al.[9] reported antimicrobial activity of fruit of C.
with Rf 0.72, which was evident as a bluish pink under fistula by agar dilution streak method at a concentration of
exposure to UV light on the reference TLC plate with zone 500 μg/ml. Only E. coli was moderately inhibited, whereas no
of inhibition 24 mm [Figure 2]. inhibition was found in case of B. subtilis and S. epidermidis.
However, in our study, B. subtilis was completely inhibited
Extracts of ethanol, methanol, and aqueous were further at concentration of 375 μg/ml by ethanol, methanol, and
tested for antimicrobial activity with time kill curve aqueous extract, while S. epidermidis was inhibited at
experiment against selective organisms. Results show concentration of 187.5 μg/ml by the same extract. Valsaraj
that ethanol, methanol, and aqueous extracts exhibit lytic et al.[15] studied the antibacterial activity of C. fistula seeds
activity against selective strain. This suggested membrane by broth dilution method and antifungal activity by ACM.
disruption as the likely mechanism of action. In case of S. According to their observation at 12.5 mg/ml concentration,
epidermidis, ethanol, methanol, and aqueous extract were E. coli and B. subtilis were inhibited while S. aureus was
bactericidal in nature, exhibiting 50% survival after 4 inhibited at a concentration 6.25 mg/ml. Perumal Samy et
hours of incubation. A er 8 hours of incubation, ethanol al.[12] reported moderate antibacterial activity of C. fistula
and methanol exhibited a prominent kill with 1.9 and 2.1% against a wide spectrum of bacteria such as E. coli, Bacillus
survival, respectively. This was followed by aqueous extract mycoides, B. subtilis, Mycobacterium smegmatis, Klebsiella
which exhibit 5.26% survival a er 8 hours incubation. This aerogenes, Pseudomonas aerogenes, and Proteus vulgaris.
concludes that ethanol is a potent bacteriostatic agent as A polar compound including 5-nonatetra contanone,

Journal of Advanced Pharmaceutical Technology & Research | Jan-Mar 2011 | Vol 2 | Issue 1 65
 Panda, et al.: Antibacterial activity of Cassia fistula leaf

Table 3: MIC and MBC of different extracts of C. fistula leaf

Plant extract Test organism Leaf Antibiotic
Gram-negative MIC MBC MBC/ TA MIC MBC
μg/ml μg/ml MIC mg/ml μg/ml μg/ml
Petroleum Escherichia coli 1 250 5 000 4.0 19.6 10 10
Ether Escherichia coli O157:H7 1 250 5 000 4.0 19.6 10 10
Salmonella typhimurium 1 250 5 000 4.0 19.6 10 10
Shigella sonnei 625 1 250 2.0 39.2 10 10
Gram-positive
Bacillus subtilis 750 1 500 2.0 32.6 10 10
Bacillus licheniformis 750 1 500 2.0 32.6 10 10
Staphylococcus aureus 625 5 000 8.0 39.2 10 10
Staphylococcus epidermidis 375 1 500 4.0 65.3 10 10
Chloroform Escherichia coli 1250 >5 000 - 12.2 10 10
Escherichia coli O157:H7 - - - - 10 10
Salmonella typhimurium - - - - 10 10
Shigella sonnei - - - - 10 10
Bacillus subtilis 750 >3 000 - 20.3 10 10
Bacillus licheniformis 750 3 000 4.0 20.3 10 10
Staphylococcus aureus 1250 >5 000 - 12.2 10 10
Staphylococcus epidermidis 750 >5 000 - 20.3 10 10
Ethanol Escherichia coli 625 5 000 8.0 253.8 10 10
Escherichia coli O157:H7 312 2 500 8.0 507 10 10
Salmonella typhimurium 1250 5 000 4.0 127 10 10
Shigella sonnei 312 1 250 4.0 507 10 10
Bacillus subtilis 187 375 2.0 848 10 10
Bacillus licheniformis 187 375 2.0 848 10 10
Staphylococcus aureus 312 625 2.0 507 10 10
Staphylococcus epidermidis 94 375 4.0 1687 10 10
Methanol Escherichia coli 625 5 000 8.0 69.4 10 10
Escherichia coli O157:H7 - - - - 10 10
Salmonella typhimurium 625 2 500 4.0 69.4 10 10
Shigella sonnei 312 625 2.0 139.1 10 10
Bacillus subtilis 375 750 4.0 115.7 10 10
Bacillus licheniformis 375 750 2.0 115.7 10 10
Staphylococcus aureus 625 2 500 4.0 69.4 10 10
Staphylococcus epidermidis 94 375 4.0 461.7 10 10
Aqueous Escherichia coli 1 250 >5 000 - 54.6 10 10
Escherichia coli O157:H7 1 250 5 000 4.0 54.6 10 10
Salmonella typhimurium 625 5 000 8.0 109.1 10 10
Shigella sonnei 1 250 5 000 4.0 54.6 10 10
Bacillus subtilis 375 1 500 4.0 181.9 10 10
Bacillus licheniformis 375 750 2.0 181.9 10 10
Staphylococcus aureus 125 250 2.0 54.6 10 10
Staphylococcus epidermidis 187 750 4.0 364.7 10 10

2-hentriacontanone, triacontane, 16-hentriacontanol, and inhibition against S. aureus compared with aqueous extract.
sitosterol along with oil showing antibacterial activity had Zones of inhibition of alcoholic and aqueous extracts were in
also been isolated in C. fistula pods.[10] Recently, Vimalraj the range of 7.0 to 12.0 mm and 7.0 to 11.6 mm, respectively.
et al.[16] tested the antibacterial activity of aqueous and MIC values of the alcoholic extracts against S. aureus were
alcoholic extract of stem bark of C. fistula with disc diffusion in the range of 0.78 to 6.25 mg/ml . Our finding was almost
and MIC methods. Alcoholic extracts recorded greater coinciding with this study.

66 Journal of Advanced Pharmaceutical Technology & Research | Jan-Mar 2011 | Vol 2 | Issue 1
 Panda, et al.: Antibacterial activity of Cassia fistula leaf

Though reports (Kumar et al., Perumal Samy et al., and cardioprotection. Phytother Res 2001;15:519-23.
Valsaraj et al.)[9,12,15] are available on antimicrobial activities of 7. Siddhuraju P, Mohan PS, Becker K. Studies on the antioxidant
seeds, pods, bark of C. fistula, no work has been conducted activity of Indian laburnum (Cassia fistula L.): A preliminary
assessment of crude extracts from stem bark, leaves, flowers and
on potentiality of leaves of this plant. Our report is of the
fruit pulp. J Agric Food Chem 2002;79:61-7.
first for antibacterial activity on leaf extracts of C. fistula.
8. Duraipandiyan V, Ignacimuthu S. Antibacterial and antifungal
S. epidermidis is a part of normal skin flora, and is o en activity of Cassia fistula L.: An ethnomedicinal plant. J
attached to the upper layer of the skin (epidermis) or Ethnopharmacol 2007;112:590-4.
mucosa, without causing any symptom. When the skin is 9. Kumar A, Pande CS. Kaul RK. Chemical examination of Cassia
injured (wounds, burns, intravenous drug addicts, etc), fistula flowers. Indian J Chem 1966;4:460
S. epidermidis may enter into deeper layers of the skin or 10. Misra TR, Singh RS, Pandey HS, Pandev RP. Chemical constituents
even the blood and cause an infection. S. epidermidis is the of hexane fraction of Cassia fistula pods. Fitoterapia 1996;57:173-4.
most common causative agent of post-cataract surgery 11. Misra TR, Singh RS, Pandey HS, Singh BK. A new diterpene from
endophthalmitis.[24] Since this bacterium is completely Cassia fistula pods. Fitoterapia 1997;58:375-7.
inhibited by leaf extracts, the plant can be useful for the 12. Perumal Samy R, Ignacimuthu S, Sen A. Screening of 34 Indian
medicinal plants for antibacterial properties. J Ethnopharmacol
treatment of S. epidermidis. 1998;62:173-82.
13. Phongpaichit S, Pujenjob N, Rukachaisirkul V, Ongsakul M.
The presence of antibacterial substances in the higher plants Antifungal activity from leaf extracts of Cassia alata L., Cassia fistula
is well established. Plants are the source of inspiration L. and Cassia tora L. Songklanakarin J Sci Technol 2004;26:741-8.
for novel drug compounds as plant-derived medicines 14. Sangetha SN, Zuraini Z, Sasidharan S, Suryani S. Antimicrobial
have made significant contribution toward human health. activities of Cassia sura ensis and Cassia fistula. J Mol Biol Biotechnol
Isolation of bioactive compounds from plant material 2008;1:1-4.
largely depends on the type of solvent used in the extraction 15. Valsaraj R, Pushpangadan P, Smi UW, Adsersen A, Nyman U.
Antimicrobial screening of selected medicinal plants from India.
procedure. The traditional healers use primarily water as J Ethnopharmacol 1997;58:75-83.
the solvent. This study concludes that the plant extraction
16. Vimalraj TR, Saravanakumar S, Vadivel S, Ramesh S, Thejomoorthy
by ethanol provided more consistent antimicrobial activity P. Antibcaterial effects of Cassia fistula extracts on pathogenic
compared with water extract. bacteria of veterinary importance. Tamilnadu J Veter Anim Sci
2009;5:109-1.
ACKNOWLEDGMENTS 17. Thatoi HN, Panda SK, Rath SK, Du a SK. Antimicrobial activity
and ethnomedicinal uses of some medicinal plants from Similipal
Biosphere Reserve, Orissa. Asian J Plant Sci 2008;7:260-7.
We wish to express our profound gratitude to Dr. A. K. Biswal
18. Barry AL. Procedure for testing antimicrobial Agents in Agar
(North Orissa University) for identification of the plant. Thanks media. In: Antibiotics in Laboratory Medicine (Lorin V, ed). Baltimore:
are also due to Dr. A. K. Bastia and Prof S. K. Du a for providing Williams Wilkins Co; 1980. p. 1-23.
necessary facilities to carry out this work and Dr. G. Sahoo for 19. Eloff JN. A sensitive and quick microplate method to determine the
his cooperation and critical suggestion on the preparation of the minimal inhibitory concentration of the plant extracts for bacteria.
manuscript. Planta Med 1998;64:711-3.
20. Panda SK, Du a SK, Bastia AK. Antibacterial activity of Croton
roxburghii balak. against the enteric pathogens. J Adv Pharm Tech
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Source of Support: Nil, Conflict of Interest: Nil.
extracts used by Sri Lankan traditional medical practitioners for

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