Phytochemical and antimicrobial activity of Tectona
grandis L.
Sandhya
Mittal1*, Nidhi Rao1, Sudhanshu1,
Ekta Menghani2
1Suresh Gyan Vihar University, Jaipur
2Mahatma Gandhi Institute of Applied Sciences, JECRC
Campus, Jaipur
ABSTRACT:
The present study has been designed with the objective to examine the
methanol extract of Tectona grandis L.
leaves (family Verbenaceae). In order to investigate
its antimicrobial potential against
strains of Shigella flexneri,
Staphylococcus aureus, Salmonella typhi,
Pseudomonas aeruginosa, Klebsiella
pneumonia, Proteus vulgaris, Enterobacter
aerogenes, Aspergillus niger, Candida albicans, Trichophyton rubrum. The methanolic extract was found to be the most effective and
showed antimicrobial activity against the organisms tested. The zone of
inhibition (mm) at various concentrations of methanolic
extract of Tectona grandis L. was found in the
range 50mg/20disc – 200mg/20disc on tested all the test organisms. The
antimicrobial activity was more significant against Salmonella typhi i.e. 21.33 mm zonetion.
The study revealed the methanol extract of Tectona grandis L. leaves against
microbes.
Key words Antibacterial
activity, Tectona grandis L. Methanolic
extract.
INTRODUCTION:
Verbenaceae is one of the major sources of antimicrobial compounds.
Tectona grandis L. is
one of the members of Verbenaceae It is called sagon saigon, saj,
taku, kayum, etc in home
languages. It grows fine in a warm climate and well-drained soil. It grows
naturally in Myanmar and central and southern India and is propagated
artificially in the Indo-Gangetic plains, and the
foothills from Bengal to Haryana and Punjab on comparatively lesser scale,
especially as an avenue tree. It grows gladly in low height plains to the
foothills up to 800 m.
Wood used to timber
merchants. Seeds used by pharmaceutical companies for medicinal preparations. Leave used for thatching, haemostatic, depurative,
anti-inflammatory, vulnery, leprosy, skin diseases, puritus, stomatities, indolent
ulcers, hemorrhages, haemoptysis, and vitiated
conditions of pitta.
The investigate for compounds with antimicrobial activity
has gained rising importance in current times, due to increasing worldwide fear
about the alarming increase in the rate of infection by antibiotic resistant
micro organisms1. However, there has also been a growing attention
in the investigation of natural products from plants for the innovation of new
anti-microbial and anti-oxidant agents in the last three decades and in recent 2,3,4,5,.
More so, a lot of of these plants have been
recognized to manufacture active secondary metabolites such as phenolic compound originate in essential oils with
recognized potent insecticidal 6 and anti-microbial activities,
which certainly has shaped the basis for their applications in some pharmaceuticals,
alternative medicines and natural therapies
Reynolds et.
al., Lis-Balchin et. al., Santos et.
al., Oloke et. al., remarked that the
world health organization has needed recognized medicinal plants as the best
source for obtaining a variety of synthetic drugs 7-10. No
hesitation, some studies have identified and remote the main active ingredients
in the plants answerable for this antimicrobial activity 11,12.
Hence, more
studies pertaining to the use of plants as therapeutic agents should be
emphasized, particularly those linked to the control of microbes. The objective
of this research is to evaluate the potential of plant extracts and Phyto-chemicals on standard Microbial bacterial and fungal
strains which are isolated from hospital. Moreover, we investigated the
synergistic effects of Tectona grandis L. with anti microbial
activity against bacteria and fungi. The purpose of this work is to evaluate
the chemical composition and antimicrobial activities of methanolic
extract of Tectona grandis L. on
the selected microorganisms.
MATERIALS AND METHODS:
Collection: Plant sample Tectona grandis L. was collected from various tribes
living in tribal pockets of Mt. Abu, arid zone of Rajasthan, in the month of
Feb, 2010. These plants were used by these tribes in their daily lives to cure
various ailments.
Identification:
These samples
were authenticated and submitted in Ethnomedicinal
Herbarium, Centre of Excellence (funded by DST), MGiaS,
Jaipur (Rajasthan).
Sources of test organisms:
Bacteria-Pure
culture of all test organisms, namely Pseudomonas
aeruginosa, Staphylococcus aureus,
Klebsiella pneumoniae,
Salmonella typhi, Shigella flexneri, Proteus vulgaris, Enterobactor aerogenes and
fungi Candida albicans,
Aspergillus niger,Trichophyton
rubrum were obtained through the courtesy of Mahatma
Gandhi Institute of applied Sciences (MGiaS), Jaipur, which were maintained on Nutrient broth media.Culture of test microbes: For the cultivation of
bacteria, Nutrient Agar Medium (NAM) was prepared by using 20 g Agar, 5 g
Peptone, 3 g beef extract and 3 g NaCl in 1 L
distilled water and sterilized at 15 lbs pressure and 121°C temperature for
25-30 min. Agar test plates were prepared to pour approximately 15 ml of NAM
into the Petri dishes (10 mm) under aseptic conditions. A saline solution was
prepared (by mixing 0.8% NaCl) in distilled water,
followed by autoclaving and the bacterial\ cultures were maintained in this
medium by regular sub-culturing and incubation at 37°C for 24-48 hour. To
prepare the test plates, in bacteria, 10-15 ml of the respective medium was
poured into the Petri plates and used for screening. For assessing the
bactericidal efficacy, a fresh suspension of the test bacteria was prepared in
saline solution from a freshly grown Agar slant.
Preparation of test extracts:
Crushed powder of
species was successively soxhlet extracted. Later,
each of the homogenates was filtered and the residue was re-extracted twice for
complete exhaustion, the extracts were sold individually. Each filtrate was
concentrated to dryness in vitro and redissolved in
alcohol, until screened for antibacterial activity.
Bactericidal and fungicidal assay:
For both, bactericidal
and fungicidal assay, in vitro Disc diffusion method was adopted13,
because of reproducibility and precision. The different test organisms were
proceeded separately using a sterile swab over previously sterilized culture
medium plates and the zone of inhibition were measured around sterilized dried
discs of Whattman No. 1 paper (6 mm in diameter),
which were contained three different concentrations, its control (of the respective
solvent) and tetracycline as reference drugs (standard disk) separately. Such
treated discs were air-dried at room temperature to remove any residual
solvent, which might interfere with the determination, sterilized and
inoculated. These plates were initially placed at low temperature for 1 hour so
as to allow the maximum diffusion of the compounds from the test disc into the
agar plate and later, incubated at 37°C for 24 hours in case of bacteria, after
which the zones of inhibition could be easily observed. Five replicates of each
test extract were examined and the mean values were then referred.
The Inhibition
Zone (IZ) in each case was recorded and the Activity Index (AI) was calculated
as compared with those of their respective standard reference drugs (AI =
Inhibition Zone of test sample/Inhibition zone of standard).
RESULT AND DISCUSSION:
Phytochemical
screening:
Phytochemical
analysis for the Tectona grandis L. The
extract was performed and the Phyto constituents
reported in the Table no. 1:
Table 1: Preliminary phytochemical
screening of the plant Tectona grandis L. [(+:
present) (- : absent)
|
Plants Extract |
Reducing sugar |
Terpe noides |
Flavo noids |
Tannin |
Saponin |
|
Methanol |
+ve |
+ve |
-ve |
+ve |
-ve |
Table
2: Antimicrobial activity of the methanol leaf extract of Tectona grandis L.
|
|
|
|
|
S.f |
S.a |
S.t |
P.v |
K. p |
P. a |
E.a |
A.n |
C.a |
T.r |
|
1. |
Methanol |
A1 |
I.Z. |
16 |
11 |
12 |
0 |
7 |
0 |
13 |
14 |
7 |
19.33 |
|
|
|
|
A.I. |
0.45 |
0.55 |
0.40 |
0 |
0.17 |
0 |
0.31 |
0.58 |
0.18 |
0.69 |
|
|
|
A2 |
I.Z. |
14 |
13.33 |
13 |
8 |
9 |
10 |
15 |
12 |
0 |
20 |
|
|
|
|
A.I. |
0.40 |
0.66 |
0.43 |
0.18 |
0.22 |
0.62 |
0.36 |
0.50 |
0 |
0.71 |
|
|
|
A3 |
I.Z. |
14 |
11 |
12 |
0 |
7 |
11 |
13 |
10 |
0 |
19 |
|
|
|
|
A.I |
0.40 |
0.55 |
0.40 |
0 |
0.17 |
0.68 |
0.31 |
0.41 |
0 |
0.67 |
|
|
|
A4 |
I.Z. |
17 |
12 |
21.33 |
8 |
8 |
11 |
13 |
7 |
8 |
14 |
|
|
|
|
A.I. |
0.48 |
0.60 |
0.71 |
0.18 |
0.20 |
0.68 |
0.31 |
0.29 |
0.21 |
0.50 |
|
|
|
A5 |
I.Z. |
13 |
10 |
13 |
0 |
8 |
13 |
12 |
12 |
10 |
17.33 |
|
|
|
|
A.I. |
0.37 |
0.50 |
0.43 |
0 |
0.20 |
0.81 |
0.29 |
0.50 |
0.27 |
0.61 |
Sf-Shigella flexneri, Sa- Staphylococcus aureus,
St-Salmonella typhi,
Pv- Proteus vulgaris, Kp- Klebsiella pneumoniae,
Pa- Pseudomonas aeruginosa, Ee-Enterobacter aerogenes,
An- Aspergillus niger, Ca-Candida albicans,
Tr- Trichophyton rubrum ; 0 - no inhibition zone.
A1-50 mg/20 disc,
A2-100 mg/20 disc, A3-150mg/20 disc, A4-200mg/20 disc, A5-250mg/20 disc. I.Z. -
Inhibition zone, I.A. – Activity index.
Antimicrobial
activity:
Antimicrobial
activity of extract from Tectona grandis L. was tested against Shigella flexneri,
Staphylococcus aureus , Salmonella typhi, Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus vulgaris,
Enterobacter aerogenes, Aspergillus niger, Candida albicans, Trichophyton rubrum by using the disc diffusion method. The plates
containing the microorganisms were then perforated and the disc was placed with
50mg/20disc,100mg/20disc,150mg/20disc,200mg/20disc,250mg/20disc concentrations.
Microbial growth was determined by measuring the diameter of the zone of
inhibition.
Figure-1- Antimicrobial activity of the methonolic
leaf extract of Tectona grandis
L.
The phytochemical analysis for the Tectona grandis L. plant
extract was performed and shown in table 1. The phyto-constituent
reported is tannin, redusing sugar, and
terpenoides and steroids and flavonoids
are not found. The antimicrobial activity of the methanolic
extract of leaf of Tectona grandis L. was studied against microorganism at various concentrations of methanolic
extract.The methanolic
extract of Tectona grandis L. at
mg concentration exhibited a significant
antimicrobial activity.
The results were
expressed in terms of the diameter of the inhibition zone in Figure 1 and Table
2: Antimicrobial efficacy of Tectona grandis (Sagwan (leaf)) methanol extract was screened against
selected test microorganisms. Methanol extract was shown appreciable inhibition
against all selected test microorganisms (bacteria and fungi). The maximum
efficacy of methanol extract A4 (200mg concentration) was against Salmonella typhi
(I.Z.-21.33mm), Shigella flexneri
(I.Z.-17mm), and A2 (100mg concentration) was active against Trichophyton rubrum
(I.Z.-20mm), Enterobacter aerogenes
(I.Z.-15mm) and A1 (50mg concentration) was also active against Trichophyton rubrum
(I.Z.-19.33mm), Shigella flexneri
(I.Z.-16mm) and A3 (150mg concentration)
showing best activity against Trichophyton rubrum
(I.Z.-19mm), Shigella flexneri (I.Z.-14mm)
and A5 (250mg concentration) showing best activity against Trichophyton rubrum (I.Z.-17.33mm), Shigella flexneri (I.Z.-13mm)
whereas all test showing good activity.
The antimicrobial
activity of Salmonella typhi was higher than
the other microorganisms. The inhibition zone diameter of S. typhi was 21.33 mm at 200mg/20disc concentration and it
was 12 mm at 50mg/20disc concentration.The plant
extract was found to have a moderate antimicrobial activity against these
microorganisms. According to the parameters, Tectona grandis L. The extract was classified as “very active” against, Salmonella
typhi and Trichophyton rubrum, “active” against Shigella flexneri, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterobacter aerogenes, Aspergillus niger,
and “partially active” against Candida albicans
and Proteus vulgaris.
CONCLUSION:
The use of plants
and their extracts in the cure of diseases back to 460-370 BC when Hippocrates
practiced the art of healing by use of plant based drugs 14. Tectona grandis L. leaf
also reported to contain carbohydrates, alkaloids, tannins, sterols, saponins, proteins, calcium, phosphorus, crude fiber and
also contain dye (yellowish-brown or reddish) 15-19. In this
connection, the present study was conducted to evaluate the antimicrobial
activity of methanol extracts of Tectona grandis L.
In this study,
the results obtained showed that the methanol extract of Tectona grandis L. inhibited the growth
of all test microorganisms. The
antimicrobial activity of Tectona grandis L. has been not studied previously. The
antimicrobial evaluation revealed that Tectona grandis L. extract was active against
all test microbial strains, being classified as very active, active and
partially active. This study is a preliminary evaluation of antimicrobial
activity of Tectona grandis L. It indicates that Tectona grandis L. have the potential to
generate novel metabolities. The plant extracts
demonstrating antimicrobial activity could result in the discovery of novel
antimicrobial agents. Besides, the same way, also be used for self medication
in domestic settings.
ACKNOWLEDGEMENT:
Author
acknowledge with thanks the financial support from Department of Science and
Technology, Government of Rajasthan, in the form of Centre with Potentials for
Excellence in Biotechnology, sanction no F 7(17) (9) Wipro/Gaprio/2006/7358-46(31/10/2008).
REFERENCES:
1. Davis
PH. Flora of Turkey and Eagean Island, Edinburg. Edinburg University, Press. 1987,
7:947.
2. Dapkevicus A, Venskutonis R, Van Beek TA, Linssen JPH. Antioxidant
activity of extract obtained by different isolation procedures from some
aromatic herbs grown in Lithuania. J.
Sci. food Agric, 1998, 77: 140-6.
3. Wang
M, Li J, Rangarajan M, Shao
Y, Lavoie EJ, Huang TC, and Ho CI. Antioxidantive phenolic compounds from sage (Salvia officinalis). Journal of Agricultural and Food Chemistry, 1998, 46:4869-4873.
4. Nascimento Grislene GF, Juliana Locatevi
Paulo C Freitas, Giuliana
LS, Antibacterial activity of plant extracts and phytochemicals
on antibiotics resistant bacteria. Brazilian
Journal of Microbiology, 2000, 31: 247-256.
5. R‘ios JL, Recio MC. Medicinal plants and antimicrobial activity. Journal of Ethanopharmacology,
2005, 100: 80-84.
6. Kambu K, Di
Phanzu N, Coune C, Waufers JN, Angenol L.
Contribution a ‘1’ etudedes properties insecticides
et, 1982, 16:34-8.
7. Reynolds
J.E.F. Martindale - The extra pharmacopeia (31st ed),
Royal pharmace
Great Britain, London, 1996.
8. Lis-Balchin M.
and Deans S.G. Bio-activity of selected plant essential oils against Listeria monocytogenes. Journal
of Applied Bacteriology, 1997, 82:759-762.
9. Santos
PRV, Oliveria ACX, Tomassini
TCB. Control Microbiological products Rev.
Faron. Bioquim, 1995,
31:35-38.
10. Oloke JK., Kolawo BO, Erhun WO. The
antibacterial and antifungal activities of certain compounds of Aframomum malegueta. Filoterapia , 1988, 59;384-8.
11. Carson
CF, Riley TV.. Antimicrobial activity of the major components of the essential
oil of Melaeuca alternifolia J. Appl. Bacteriol,
1995, 96:264-9.
12. Fabricant
DS, Famsworth NR. The value of plant used in
traditional medicine for drug discovery, Environmental
Health Perspectives Supplements, 2001, 109:69-75.
13. Gould
JC, and Bowie JH. The determination of bacterial sensitivity of antibiotics. Edinburgh Med. J., 1952, 59: 178-199.
14. Sofowora EA.
Medicinal plants and traditional medicine in Africa. John Wiley and Sons, Chichester, 1982, PP
198.
15. The Ayurvedic Pharmacopoeia of India, part-1, Vol-1, First
edition. , 2006, 137-143.
16. Goswami DV, Nirmal SA, Patil MJ, Dighe NS, Laware RB and Pattan SR. An over view of Tectona grandis: Chemistry and Pharmacological
profile. Phcog Rev, 2009, 3(5): 170-174.
17. Krishnan
marg, K. S., 2006, “The Useful plants Of India”:
62322.
18. Majumdar M, Nayeem N, Kamath JV, Asad M. Apr, Evaluation of Tectona grandis leaves for wound healing
activity, Pak J Pharm
Sci., 2007, 20(2):120-4.
19. Naira Nayeem Karvekar MD. “Analgesic
and anti-inflammatory activity of the methanolic
extract of the frontal leaves of Tectona grandis. The
Internet Journal of Pharmacology, 2010, 8(1): 1531-2976.
Received on 15.09.2012
Modified on 03.10.2012
Accepted on 09.10.2012
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Research J. Science and Tech. 4(5):
September –October, 2012:
188-191