Antimicrobial and Antifungal Activity of Centella asiatica (L.)Urban, Umbeliferae

 

NS Jagtap*, SS Khadabadi, DS Ghorpade, NB Banarase, SS Naphade

Government College of Pharmacy, Pharmacognosy and Phytochemistry Department, Kathora Naka, Amravati- 444 604 (M.S.)

*Corresponding Author E-mail: nikejagtap@gmail.com

 

ABSTRACT

Centella asiatica (Linn.) urban belonging to family Umbeliferae popularly knows as “Brahmi “,  is very useful medicinal plant described by Charaka as an anti-aging plant. An experiment was carried out to study the antimicrobial activity of petroleum ether, ethanol and water extract of Centella asiatica plant by agar diffusion method. Zone of inhibition produced by petroleum ether, ethanol and water extract in dose of 62.5, 125,250,500 and 1000 µg/ml against some selected strains was measured and compared with standard antibiotics ciprofloxacin (10µg/ml). The present study demonstrated that the ethanolic extract of Centella asiatica has higher antimicrobial activity than petroleum ether and water extract.

 

KEYWORDS:  Centella asiatica, antimicrobial activity, agar diffusion method.

 

INTRODUCTION:

Many plants were found to contain compounds, which are used as natural medicines to treat common bacterial infections. Indian medicinal plants are regularly used in various system of medicine because of minimal side effect and cost effectiveness. The potential for developing antimicrobials from higher plants appears rewarding as it may lead to the development of phytomedicine against microbes. Hence the sensitivity study of bacterial strains to the plant Centella asiatica was evaluated.           

              

Centella asiatica (Linn.) urban belonging to family Umbeliferae popularly knows as “Brahmi “, is very useful medicinal plant described by Charaka as an anti-aging plant. It is creeping plants, but leaves are bigger and long petiolate1. The leaves are entire, crenate, orbicular and reniform. Leaves are 1.5-6.5 cm in diameter, petioles 7.5-15 cm in length, stipules are short forming sheathing base. It bears an umbel inflorescence with 3-4 pink sessile flowers. The stems are red and show long internodes. This plant is found in marshy areas all over India, Sri Lanka, Madagascar and Africa up to an altitude of 650 m.

          

Various chemical constituents are reported in Centella asiatica like asiaticoside, madecassoide, madecassic acid, asiatic acid, glucose, rhamnose, terpenoids , sitosterol, stigmasterol, fatty oil consist of glycerides of palmitic acid, stearic acid, linoleic acid, linolenic acid, ascorbic acid, . It also contains calcium, iron, and phosphate2,3.

 

It is mainly used to treat mental and neurological disturbances. Apart from its role as a brain tonic, its efficacy in treating tuberculosis, syphilis, amoebic dysentery and common cold is well documented in the literature. Also used in skin disease, leprosy, wound healing, anti-ulcer activity, mild diuretics4.   

 

MATERIAL AND METHODS:

Plant Material:

The aerial part of plant of Centella asiatica was collected from local area of Amravati in month of Novembers – December 2008. It was identified and authenticated by taxonomist Dr. Prabha Bhogaonkar, Director of Govt. Vidarbha Institute of Science and Humanities, Amravati, and Maharashtra, India.

 

Extraction of plant material:

The aerial part of plant of Centella asiatica was dried in shade and crushed to fine powder. The extraction of plant was done by petroleum ether (60-800 C), ethanol and water by using Soxhlet apparatus. The solvent was evaporated and the extract obtained was cooled and dried under vacuum. Then extract is dissolved in DMSO for antimicrobial activity.


TABLE 1:  Antimicrobial activity of petroleum ether extract of Centella asiatica

Microorganism

Zone of Inhibition (mm)  (µg/ml)

MIC (µg/ml)

Ciprofloxacin  (10µg/ml)

62.5

125

250

500

1000

P.vulgaris (BS)

00

10

14

15

17

125

24

S.aureus(BS)

00

00

10

14

16

250

22

E. coli(BS)

00

00

00

12

15

500

20

B. subtilis(BS)

09

10

14

17

19

62.5

26

A. flavus(FS)

00

11

12

16

18

125

24

A. niger(FS)

00

10

11

13

16

125

22

C. albicans(FS)

00

09

11

12

15

125

20

BS- Bacterial Strain; FS- Fungal strain

 

TABLE 2: Antimicrobial activity of Ethanolic extract of Centella asiatica

      Bacteria

          Zone of Inhibition  (mm)  (µg/ml)

  MIC (µg/ml)

Ciprofloxacin  (10µg/ml)

62.5

125

250

500

1000

P.vulgaris (BS)

00

12

16

17

20

125

24

S.aureus(BS)

00

08

12

17

20

125

22

E. coli(BS)

00

06

10

14

18

125

20

B. subtilis(BS)

10

13

16

18

22

62.5

26

A. flavus(FS)

09

12

13

16

21

62.5

24

A. niger(FS)

00

11

12

14

18

125

22

C. albicans(FS)

00

09

12

12

16

125

20

BS- Bacterial Strain; FS- Fungal strain

 

 

Bacterial and fungal strains used:

Standard antibiotics ciprofloxacin (10µg/ml). Bacterial strains Staphylococcus aureus (NCIM 2079), Escherichia coli (NCIM 2065), Bacillus subtilis (NCIM 2063), Propionibacterium vulgaris (NCIM 2813), and the fungal strains like Aspergillus niger (NCIM 596), Aspergillus flavus (NCIM 519), Candida albicans (NCIM 3100), were obtained from National Collection of Industrial. Microorganism (NCIM), Pune, India. Dimethyl sulphoxide (DMSO) was purchased from Sigma Aldrich. All other chemicals used were of high analytical grade

 

Culture Medium and Inoculum:

The stock cultures of microorganism used in this study were maintained on Plate Count Agar slants at 40 C. Cell suspension were prepared by inoculation of each bacteria into 10 ml of Nutrient buyyon (Difco).  Incubation was performed at 37 0 C for 24 hr. On the next day Mueller- Hinton Agar (MHA) and Sabouraud Dextrose Maltose Hiveg Agar (SDMA) was prepared and cooled at 450C. Bacterial and fungal suspension was added into MHA and SDMA respectively to give a final concentration of 107 microorganism/ ml and plate out9.

 

Antimicrobial activity 6-13:

A previously liquefied medium of agar nutrient was inoculated with the suspensions of bacteria, the suspension is added to the sterile Mueller-Hinton Agar medium at a temperature between 40-500C and the inoculated medium was poured immediately into petri-dishes to occupy a depth of 3-4 mm. The prepared dishes must be stored such that no significant growth or death of the test bacteria occurs before used and the surface of the agar layer was dry at the time of used. Cups cut in medium using sterile cork borer about 8 mm in diameter.

 

The cut agar disc was removed by vacuum device. Then standard antibiotics ciprofloxacin (10µg/ml) and five different dilutions(62.5,125,250,500,and 1000 µg/ml) of the petroleum ether, ethanol and water extract place in appropriate position on the plate with quadrants marked at the back of the each petri dishes. The petri dishes were incubated at 37oC for 24 hrs and the diameter of the zone of inhibition measured in mm (including 8mm diameter of wells). The activity of extract was compared with

 

standard ciprofloxacin (10µg/ml). The minimum dimension of the zone of no microbial growth and minimum inhibitory concentrations (MIC) were determined. An independent determination of three different extract was recorded for the bacteria and fungi. The result of antimicrobial activity of petroleum ether extract shows in Table1, ethanolic extract shows in Table-2, water extract shows in Table 3.

 

RESULT AND DISCUSSION:

Present research work deals with the evaluation of three different extract of plant of Centella asiatica for antimicrobial activity. In this research work petroleum ether, ethanol and water extract were prepared. The antimicrobial activity was performed by using agar diffusion method.

             

As per data obtained the growth of inhibition of four bacterial strains and three fungal strains varied largely, B.subtilis shows maximum rate of inhibition compare to other bacterial strains and A. flavus shows maximum rate of inhibition compare to other fungal strains . While other bacterial and fungal strains shows the moderately zone of inhibition. It was examined from the available

 

 

TABLE 3: Antimicrobial activity of Water extract of Centella asiatica

Bacteria

Zone of Inhibition (mm)    (µg/ml)

MIC (µg/ml)

Ciprofloxacin   (10µg/ml)

62.5

125

250

500

1000

P.vulgaris (BS)

00

08

12

13

16

125

24

S.aureus(BS)

00

00

06

11

13

250

22

E. coli(BS)

00

00

00

11

14

500

20

B. subtilis(BS)

06

09

13

15

17

62.5

26

A. flavus(FS)

00

09

09

11

12

125

24

A. niger(FS)

00

00

11

12

14

250

22

C. albicans(FS)

00

09

10

12

13

125

20

BS- Bacterial Strain; FS- Fungal strain

 

 

data (Table 1, 2 and 3) that ethanolic extract of Centella asiatica shows significantly higher rate of sensitivity against various bacteria strains, while petroleum ether shows moderately sensitivity and water showed that least sensitivity.

 

ACKNOWLEGEMENT:

The author is thankful to Mrs. Sharda Deore for his constant support throughout research work.

 

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Received on   .  .2008       Modified on   .  .2008

Accepted on   .  .2008       © RJPT All right reserved

Research J. Pharm. and Tech.2 (2): April.-June.2009; Page 328-330