EVALUATION OF ANTIMICROBIAL AND PHYTOCHEMICAL PROPERTIES OF TERMINALIA ARJUNA LINN. EXTRACTS AGAINST RESPIRATORY TRACT PATHOGENS

Abstract

The aim of was aimed to investig causing respiratory infections. Dried plant materials were crushed and extracted in petroleum ether, acetone, methanol and water by using to examine ant for plant extract also. The results showed that other extract. The maximum inhibition was found against by H. influenzae aureus (12.6±0.28 Phytochemical and tannins in plant extracts. respiratory diseases which might be accountable for its antimicrobial potential.

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EVALUATION OF ANTIMICROBIAL AND PHYTOCHEMICAL PROPERTIES OF
LINN. EXTRACTS AGAINST RESPIRATORY TRACT PATHOGENS
1
*Sanjay Kumar,
1
Department of Botany and Microbiology, Gurukul Kangri University, Haridwar
2
Department of Botany and Microbiology, H.N.B. Garhwal University, Srinagar
ARTICLE INFO
ABSTRACT
The aim of
was aimed to investig
causing respiratory infections. Dried plant materials were crushed and extracted in petroleum ether,
acetone, methanol and water by using
to examine ant
for plant extract also. The results showed that
other extract. The maximum inhibition was found against
by H.
influenzae
aureus
(12.6±0.28
Phytochemical
and tannins in plant extracts.
respiratory diseases which might be accountable
Copyright © 2014 Sanjay Kumar et al. This
is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
INTRODUCTION
Herbal medicines have been used for the treatment and
various diseases in
Indian traditional practiced methods such as
Ayurveda, Unani and Siddha (Dahanukar
et al.,
et al., 2006). Terminalia arjuna (
Combretaceae
called arjuna in Hindi. It
is a deciduous and evergreen tree
distributed throughout India and also found in Burma, Srilanka
and Mauritius, growing up to a height of 60 to 90 feet. Leaves
of arjuna are simple, oblong or elliptic with pale and dark green
upper surface and pale brown lower surface. Flowers are
bisexual, sessile and white arranged in short axillary spikes or
in te
rminal pannicule. The bark is smooth, pinkish
outside and flakes off in large, curved and rather flat pieces
(Dwivedi et al., 2007; Jain et al
., 2009). The bark and leaves of
this plant have been used in indigenous system of medicine for
curing d
ifferent diseases, the bark in the treatment for angina
(hritshool), expectorant, antidysentric, purgative, laxative,
leucoderma, anaemia, hyperhidrosis, asthama, tumors and other
cardiovascular disorders (Udupa, 1986).
Literature search
revealed that bark possesses
good anticancer, antiviral and
antimicrobial activities (Tripathi and Singh, 1996; Cooper,
2005; Singh et al., 2008).
*Corresponding author: Sanjay Kumar,
Department of Botany and Microbiology, Gurukul Kangri University,
Haridwar-249404, Uttarakhand, India.
ISSN: 0975-833X
Available online at http://www.journal
International Journal of Current Research
Vol.
Article History:
Received 14
th
May, 2014
Received in revised form
18
th
June, 2014
Accepted 07
th
July, 2014
Published online 31
st
August,
2014
Key words:
Antimicrobial activity,
Agar well diffusion method,
Terminalia arjuna,
Respiratory diseases.
RESEARCH ARTICLE
EVALUATION OF ANTIMICROBIAL AND PHYTOCHEMICAL PROPERTIES OF TERMINALIA ARJUNA
LINN. EXTRACTS AGAINST RESPIRATORY TRACT PATHOGENS
*Sanjay Kumar,
2
Kishlay Kumar and
1
Navneet
Department of Botany and Microbiology, Gurukul Kangri University, Haridwar
-
249404, Uttarakhand, India
Department of Botany and Microbiology, H.N.B. Garhwal University, Srinagar
- 246274,
ABSTRACT
The aim of
this research was to evaluate the antimicrobial activity of T
erminalia a
was aimed to investig
ate against three gram-positive, two gram-
negative bacteria and one fungi
causing respiratory infections. Dried plant materials were crushed and extracted in petroleum ether,
acetone, methanol and water by using
Soxhlet
apparatus. The agar well diffusion method was adopted
to examine ant
imicrobial activity of
extracts against test organisms. Phytochemical analysis was done
for plant extract also. The results showed that
methanol
extract was most active as comparison to
other extract. The maximum inhibition was found against
S.
pneumoniae
influenzae
(16.6±0.57mm), P. aeruginosa (14.6±0.76 mm) and
S. pyogenes
(12.6±0.28
mm) and minimum against C. albicans
(11.3±0.28 mm) respectively.
Phytochemical
screening revealed the presence of alka
loids, flavonoids, glycosides, steroids, saponins
and tannins in plant extracts. The results signify traditional values of
respiratory diseases which might be accountable
for its antimicrobial potential.
is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
Herbal medicines have been used for the treatment and
cure of
Indian traditional practiced methods such as
et al.,
2000; Kumar
Combretaceae
) is commonly
is a deciduous and evergreen tree
distributed throughout India and also found in Burma, Srilanka
and Mauritius, growing up to a height of 60 to 90 feet. Leaves
of arjuna are simple, oblong or elliptic with pale and dark green
upper surface and pale brown lower surface. Flowers are
bisexual, sessile and white arranged in short axillary spikes or
rminal pannicule. The bark is smooth, pinkish
-grey from
outside and flakes off in large, curved and rather flat pieces
., 2009). The bark and leaves of
this plant have been used in indigenous system of medicine for
ifferent diseases, the bark in the treatment for angina
(hritshool), expectorant, antidysentric, purgative, laxative,
leucoderma, anaemia, hyperhidrosis, asthama, tumors and other
Literature search
good anticancer, antiviral and
antimicrobial activities (Tripathi and Singh, 1996; Cooper,
Department of Botany and Microbiology, Gurukul Kangri University,
Plants have unlimited capacity to synthesize secondary
metabolites such as tannins, terpenoids, alkaloids, flavo
glycosides and phenols which have been found to have
antimicrobial properties (
Cowan, 1999; Sher, 2009; Das
2010).
The purpose of this research was to determine the
antimicrobial and phytochemical aspects of
extracts against respiratory pathogens that usually cause
infections in upper and lower respiratory tract region.
MATERIALS
AND METHODS
Plant collection
Plant was collected from Srinagar, Uttarakhand and
authenticated at Department of Botany and Microbiology,
H.N.B. Garhwal
University, Srinagar. Collected roots were
dried under shade at room temperature and crushed to small
pieces by using pestle
and mortar and powdered in an electric
grinder.
Extraction of plant material
Plant extracts were prepared by immersing 200 g of
plant material in 600 ml of four different solvents i.e.
petroleum ether (PET), acetone (ACE), methanol (MeOH) and
water (H
2
O), loaded in Soxhlet assembly and extracted for 72 h
through successive method (Ahmed
et al
were filtered through Whatman No. 1 filter paper and crude
Available online at http://www.journal
cra.com
International Journal of Current Research
Vol.
6, Issue, 08, pp.8003-8005, August,
2014
INTERNATIONAL
OF CURRENT RESEARCH
z
TERMINALIA ARJUNA
LINN. EXTRACTS AGAINST RESPIRATORY TRACT PATHOGENS
249404, Uttarakhand, India
Uttarakhand, India
erminalia a
rjuna bark extracts
negative bacteria and one fungi
causing respiratory infections. Dried plant materials were crushed and extracted in petroleum ether,
apparatus. The agar well diffusion method was adopted
extracts against test organisms. Phytochemical analysis was done
extract was most active as comparison to
pneumoniae
(17.3±0.57 mm) followed
S. pyogenes
(13.6±0.28 mm), S.
(11.3±0.28 mm) respectively.
loids, flavonoids, glycosides, steroids, saponins
The results signify traditional values of
T. arjuna in treatment of
for its antimicrobial potential.
is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use,
Plants have unlimited capacity to synthesize secondary
metabolites such as tannins, terpenoids, alkaloids, flavo
noids,
glycosides and phenols which have been found to have
Cowan, 1999; Sher, 2009; Das
et al.,
The purpose of this research was to determine the
antimicrobial and phytochemical aspects of
T. arjuna bark
extracts against respiratory pathogens that usually cause
infections in upper and lower respiratory tract region.
AND METHODS
Plant was collected from Srinagar, Uttarakhand and
authenticated at Department of Botany and Microbiology,
University, Srinagar. Collected roots were
dried under shade at room temperature and crushed to small
and mortar and powdered in an electric
Plant extracts were prepared by immersing 200 g of
powdered
plant material in 600 ml of four different solvents i.e.
petroleum ether (PET), acetone (ACE), methanol (MeOH) and
O), loaded in Soxhlet assembly and extracted for 72 h
et al
, 1998). Plant extracts
were filtered through Whatman No. 1 filter paper and crude
INTERNATIONAL
JOURNAL
OF CURRENT RESEARCH

extracts obtained by removing solvent in rotary vacuum
evaporator. Residues were stored at 4°C until further use.
Test Microorganisms
Five standard bacterial strains (Haemophilus influenzae MTCC
3826, Pseudomonas aeruginosa MTCC 2474, Staphylococcus
aureus MTCC 1144, Streptococcus pneumoniae MTCC 655,
Streptococcus pyogenes MTCC 442) and one fungi Candida
albicans MTCC 227 and their isolates were subjected for
antimicrobial study. The selected microbial standard strains
were procured from Institute of Microbial Technology
(IMTECH), Chandigarh.
Screening for antimicrobial activity
The antimicrobial activity of different extracts was determined
by agar well-diffusion method (Perez et al., 1990). 0.1 ml of
12-16 h incubated cultures of microbial species were mixed in
molten Mueller Hinton Agar (MHA) medium and poured in
pre-sterilized Petri plates. A cork borer (6 mm diameter) used
to punch wells in solidified medium and filled with extracts of
45 μl of 200 mg/ml final concentration of extracts. DMSO was
used as negative control. The efficacy of extracts against test
organisms was compared with broad spectrum antibiotic
erythromycin (positive control). The plates were incubated at
37°C for 24 to 48 h in BOD incubator and the diameter of the
zone of inhibition was measured in millimetre. Each sample
was assayed in triplicate and mean values were observed. The
antimicrobial activity was interpreted from size of diameter of
zone of inhibition measured to nearest millimetre (mm) as
observed from clear zones surrounding wells.
Phytochemical screening
Major phytoconstituents present in T. arjuna extracts were
subjected to phytochemical analysis to determine the presence
of bioactive components by using standard qualitative methods
(Trease and Evans, 1996).
Test for alkaloids
Test solution was acidified with acetic acid and a drop of
Mayer’s reagent was added. A white precipitate indicated the
presence of alkaloids.
Test for flavonoids
On addition of conc. HCl in MeOH extract of material, a red
colour appeared which indicated the presence of flavonoids.
Test for glycosides
Plant extract was filtered and sugar was removed by
fermentation with baker’s yeast. The acid was removed by
precipitation with Ba(OH)2. The remaining extract contained
the glycosides. The hydrolysis of solution was done with conc.
H2SO4 and after hydrolysis the presence of sugars was
determined with help of Fehling’s solution.
Test for Steroids
Extracts were mixed with 3 ml CHCl3 and 2 ml conc. H2SO4
was poured from side of test tube and colour of the ring at
junction of two layers was noted. A red colour showed the
presence of steroids.
Test for Saponins
Extracts were diluted with distilled water to 20 ml and this was
shaken in a graduated cylinder for 15 minutes. Formation of 1
cm layer of foam indicates the presence of saponins.
Test for Tannins
Extract was added in 1% ferric chloride and observed the
colour. Bluish black colour appeared which disappeared on
addition of dilute H2SO4 follow a yellow brown precipitate
indicates the presence of tannins.
RESULTS AND DISCUSSION
The results for antimicrobial activity are depicted in Table 1.
MeOH extract was found most active against all test pathogens
in comparison to other extracts.
MeOH extract showed maximum activity followed by PET,
ACE and H2O extract. The maximum inhibition was found
against S. pneumoniae (17.3±0.57 mm) followed by H.
influenzae (16.6±0.57mm), P. aeruginosa (14.6±0.76 mm) and
S. pyogenes (13.6±0.28 mm), S. aureus (12.6±0.28 mm)
respectively. The minimum inhibition was noted against C.
Table 1. The percentage of potency of
Terminalia arjuna
extracts against respiratory tract pathogens
S. No.
Pathogens Diameters of inhibition zone (mm)
PET CHCl
3
MeOH H
2
O
1. H. influenzae 9.0±0.50 13.3±0.57 15.3±0.28 11.0±0.50
2. H. influenzae (MTCC 3826) 10.3±0.57 12.3±0.57 16.6±0.57 12.3±0.28
3. P. aeruginosa 8.3±0.28 12.3±0.28 14.0±0.50 11.3±0.28
4. P. aeruginosa (MTCC 2474) 9.6±0.76 11.3±0.57 14.6±0.76 11.0±0.50
5. S. aureus 8.6±0.28 11.3±0.28 12.3±0.28 12.3±0.57
6. S. aureus (MTCC 1144) 9.6±0.28 10.6±0.28 12.6±0.28 13.6±0.28
7. S. pneumoniae 9.6±0.76 13.6±0.76 17.3±0.57 11.3±0.57
8. S. pneumoniae (MTCC 655) 9.3±0.57 15.0±0.50 16.3±0.57 10.3±0.28
9. S. pyogenes 10.0±0.50 11.3±0.57 13.6±0.28 12.6±0.76
10. S. pyogenes (MTCC 442) 9.3±0.28 10.3±0.28 12.6±0.76 13.0±0.50
11. Candia albicans (MTCC 227) 7.6±0.28 9.3±0.57 11.3±0.28 9.6±0.76
*Values are means of three replicates, Cork borer diameter: 6 mm
8004 Sanjay Kumar et al. Evaluation of antimicrobial and phytochemical properties of terminalia arjuna linn. extracts against respiratory tract pathogens

albicans (11.3±0.28 mm). So far, antimicrobial potential of T.
arjuna has been found effective against selected bacterial and
fungal species. T. arjuna ACE leaf extract was found most
effective against S. aureus (28 mm) followed by Proteus
mirabilis (27.6 mm), Acinetobacter sp. (16.6 mm) and P.
aeruginosa (16 mm) (Aneja et al., 2012). Different pathogenic
bacteria such as Escherichia coli, Klebsiella pneumoniae, and
Salmonella spp. are widely distributed in the hospitals and in
community thus creating serious health problems (Khan, 2004;
Akram et al., 2007).
Table 2. Phytochemical screening of various extracts of T. arjuna
bark
S. No. Phytoconstituents Solvents
PET ACE MeOH H
2
O
1. Alkaloids + + + +
2. Flavonoids - + + +
3. Glycosides - + - +
4. Steroids + + + +
5. Saponins - - + +
6. Tannins + + + +
+ = Present, - = Absent
The phytochemical screening of T. arjuna extract has shown
that plant contains alkaloids, flavonoids, glycosides, steroids,
saponins and tannins which are very important constituent
when looking for pharmacologically active phytochemical in
the plant (Table 2). According to Akhter et al. (2012), extracts
of T. arjuna (bark) contain phenols, flavonoids, tannin,
saponin, alkaloids, glycosides, phytosterols and carbohydrate.
Doorika et al. (2012), reported T. arjuna were screened of
biologically active compounds like steroids, tannins, phenolics
compound, quinone, terpinoids, sugar, alkaloids and
flavonoids. By this study, it is concluded that T. arjuna can be
used as herbal medicine to treat respiratory infections caused
by tested pathogens as comparative to synthetic
chemotherapeutic agents. It is urged that further research
should be carried out to expose the bioactive constituents
present in T. arjuna.
Conclusion
The investigation for antimicrobial activity of T. arjuna bark
extracts revealed that plant has broad spectrum activity against
selected pathogens which explain the basis for its use in
traditional medicines. The significant activity was exhibited by
MeOH extract against test respiratory pathogenic
microorganisms. The study supported the usefulness in term of
availability of phytoconstituents. By results, it can be
concluded that T. arjuna can be helpful as an alternative source
of medicine and new drug discovery.
Acknowledgement
The authors are thankful to Head, Department of Botany &
Microbiology, Gurukul Kangri University, Haridwar to provide
necessary laboratory facilities to pursue this research work.
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8005 International Journal of Current Research, Vol. 6, Issue, 08, pp.8003-8005, August,
2014