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).

 

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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