INTERNATIONALE PHARMACEUTICA SCIENCIA

| April-June 2011 | Vol. 1 | Issue 2 |

Available online http://www.ipharmsciencia.com
ISSN 2231-5896
2011 IPS
RESEARCH ARTICLE

Antimicrobial properties of Abelmoschus manihot Linn

ABSTRACT

Jain Pritam*, Bari Sanjay1

Antimicrobial activity of woody stem of Abelmoschus manihot, were tested

against different bacteria Escherichia coli, Staphylococcus aureus, Bacillus subtilis,
Klebsiella pncumoniae,, Pseudomonas aeruginosa, Bacillus pumilis and fungi
Candida albicans. Aspergillus niger, Fusarium monoliforme, Trichoderma
viridae, Phanerochaete chrysosporium and Pcilomyces species by cup plate
diffusion method. Minimal Inhibitory Concentration (MIC) values of each active

*Associate Professor, 1Professor

Department of Pharmaceutical
Chemistry,
R. C. Patel Institute of
Pharmaceutical Education and
Research,
Karwand Naka, Shirpur Dist. Dhule
425405 Maharashtra India

extract were determined. The chloroform and methanol extracts exhibited

prominent activity against bacteria like Escherichia coli, Klebsiella
pneumoniae, Pseudomonas aeruginosa and fungi like Aspergillus niger

Date of Submission: 16-06-2011

Date of Acceptance: 22-06-2011
Conflict of interest: Nil
Source of support: None

and Trichoderma viridae as compared to petroleum ether extract.

Key words: Antimicrobial activity, Abelmoschus manihot, zone of
inhibition, minimum inhibitory concentration.
Introduction:

1.8 m. high. It is Native to China, introduced into

Infectious diseases represent a critical problem to health and

India,

they are one of the main causes of morbidity and mortality

Maharashtra. The flowers used in the treatment of

worldwide. During the past several years, there has been

chronic bronchitis and toothache. The ethanol extract

an increasing incidence bacterial and fungal infections due

of flower was screened for antiviral activity, and it was

to a growth in irnmunocompramised population such as

observed that the hyperoside shown significant anti

organ transplant recipients, cancer, and HIV/AIDS

HBV activity [3]. The leaves and roots showed

patients. The changing pattern of clinical evaluation and

significant wound healing activity. The leaves were

regulatory requirements for merits and demerits of drugs will

tested on bone loss in ovarectomised rats and it was

be highlighted for future challenges and advances in

observed that it was able to prevent the ovariectomy

antimicrobial drug development. Resurgence in the use of

induced femoral osteopenia [4].

near

Calcutta

and

in

coastal

areas

of

herbal medicines worldwide has provided an excellent

opportunity to Indian companies to look for therapeutic

Materials and methods:

leads from Indian ancient system of ayurveda that could be

Plant material

utilized for drug development [1]. Abelmoschus manihot

The fresh woody stem of Abelmoschus manihot

(Malvaceae)[2] is a large annual erect hairy plant, 1.2-

collected from Toranmal hills of Maharashtra, India in

June 2010 and authenticated by Dr. D. A. Patil, HOD

Address for correspondence

Botany Dept, SSVPS College, Dhule, Maharashtra,

Associate Professor
Dept. of Pharmaceutical Chemistry,
R. C. Patel Institute of Pharmaceutical Education and
Research, Karwand Naka, Shirpur Dist. Dhule 425405
Maharashtra India
Email: pritash79@yahoo.com, sbbari@rediffmail.com
32

India. The woody stem was shed dried, ground and sieved
with a 40 mesh sieve.
Preparation of extracts

Internationale Pharmaceutica Sciencia

Apr-Jun 2011

Vol 1 Issue 2

Jain Pritam et al: Antimicrobial properties of Abelmoschus manihot Linn

kg of stem powder was subjected to hot

antibacterial agent Amoxycillin (10 g/disc) and

extraction using soxhlet extractor, successively with

antifungal agent Ketoconazole (50 g/disc) were

petroleum ether, chloroform and methanol. All the extracts

prepared. Each plate was inoculated with 20m1 of

were concentrated under reduce pressure by using rotary

microbial suspension having a concentration of Ix 10s

flash vacuum evaporator and then dried by using vacuum

cells/ml. 0.1 ml of extract was added to each cup. The

dryer, giving PEA (0.65%), CEA (0.73%), and MEA (

plates containing bacteria were incubated at 37 C for

2.50%) respectively.

24h and those containing fungi were incubated at 25 C

About 4

for 7 days. The positive antimicrobial activity was read

Test microorganisms

based on growth inhibition zone and compared with

Strains, including fungi and bacteria were obtained from

the standard drug. In order to determine the minimum

National Chemical Laboratories (NCL), Pune Maharashtra.

inhibitory

Escherichia coli NCIM 2110, Staphylococcus aureus

minimum concentrations of agents showing growth

NCIM 2079, Bacillus subtilis NCIM 2250, Klebsiella

inhibition zone when examined visually, extracts were

pneumoniae NCIM 2719, Pseudomonas aeruginosa

dissolved in DMSO to make a concentration of 100

NCIM 2036, Bacillus pumilis NCIM 2327 and

mg/ml. The extracts were diluted in a simple dilution

Candida albicans NCIM 3471, Aspergillns niger NCIM

manner to make concentrations in the range of 0.15,

545, Fusarium monoliforme NCIM 1099, Trichoderma

0.31, 0.62, 1.25 and 2.5 mg/ml. 0.1 ml of the extracts

viridae NCIM 1221, Phanerochaete chrysosporium

was then added to each cup. All the tests were repeated

NCIM 1197 and Pcilomyces species NCIM 1081 were

in triplicates.

concentration

values,

which

are

the

used as test organisms.

Phytochemical studies:
Method

of preparation of

test

organism

Our phytochemical investigations of chloroform extract

suspension

revealed the presence of terpenes and steroids viz.

Test organism maintained on slants of medium containing

stigmasterol, campesterol, and beta sitosterol while

300mg of manganese sulphate per liter and transferred to

methanol extract shows the presence of flavonoids

fresh slant once a week. Then the slants incubated at

viz. dodecanol and butanedioic acid methyl ester [6].

temperature 32CC for 24 hours. Organism was washed by

using 3 ml of saline solution from agar slant onto a large

Results and Discussion:

agar surface of medium such as Roux bottle containing 250

As shown in Table 1 and 2, the chloroform and methanol

ml of agar. It was incubated for 24 hour. Using 50 ml

extracts exhibited prominent activity against bacteria

saline solution, the growth from the nutrient surface

like

was

under

Pseudomonas aeruginosa and fungi like Aspergillus

refrigeration. Inoculum was adjusted at 530 nm, which

niger and Trichoderma viridae as compared to

give transmission equivalent to Ix 10s cells/ ml.

petroleum ether extract.

Antimicrobial Assay:

Conclusion:

Antimicrobial activity of the above mentioned extracts

From the results, it can be concluded that the

was determined, using a modified cup plate method [5].

Abelmoschus manihot woody stem extracts find use as

Muller Hinton agar was used for the growth of

broad-spectrum antimicrobial agent after extensive

bacterial strains and Potato Dextrose agar was used

investigation. These results may provide a basis for the

for the growth of fungi, in case of spore producing

isolation of compounds of biological interest from

organism, sporulated culture was also grown on Potato

Abelmoschus manihot for potent activity.

washed.

Then

organism

stored

Escherichia

coli,

Klebsiella

pneumoniae,

Dextrose agar. Plant extracts were dissolved in DMSO

at a concentration of 500 g/ml and standard
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Jain Pritam et al: Antimicrobial properties of Abelmoschus manihot Linn

Table 1
Zone of inhibition in diameter (mm) of Abelmoschus manihot woody stem extracts by agar well diffusion method.
Microorganisms

Microorganisms

Pet ether

Chloroform

Methanol

Standard Drug

Bacteria
Escherichia coli(NCIM 2110)

10.4

11.4

11.9

15.0

Staphylococcus aureus (NCIM 2079)

9.8

11.2

11.2

14.3

Bacillus subtilis (NCIM 2250)

8.9

10.7

10.5

16.4

Pseudommonas aeruginosa (NCIM 2036)

10.0

11.6

11.7

17.0

Klebsiella pneumoniae (NCIM 2719)

10.1

11.1

11.6

15.2

Bacillus pumilis (NCIM 2327)

10.2

10.3

10.5

16.3

Fungi
Candida albicans (NCIM 3471)

9.6

11.6

11.2

14.8

Aspergillus niger (NCIM 545)

8.4

11.0

11.8

16.7

Fusarium monoliforme (NCIM 1099)

8.9

11.0

11.4

17.1

Trichoderma viridae (NCIM 1221)

9.3

11.1

12.0

18.0

Phanerochaete chrysosporium (NCIM 1197)

10.0

10.0

10.8

14.5

9.7

10.1

10.5

14.2

Pcilomyces species (NCIM 1081)

Values are inhibition zone (mm), and an average of triplicate.

Each extract has concentration of 500 g/ml,
Standard Drugs: Amoxycillin (10 g/disc), Ketoconazole (50 g/disc)
Incubation conditions for bacteria1 day at 370 C. For fungi7 days at 270 C.

Table 2
Minimum Inhibitory concentration (mg/ml) of Abelmoschus manihot woody stem extracts by tube dilution
method
MIC (mg/ml)

Microorganisms

Pet ether

Chloroform

Methanol

Standard Drug

Escherichia coli(NCIM 2110)

2.5

1.25

0.62

0.24

Staphylococcus aureus (NCIM 2079)

1.25

0.62

0.15

0.24

Bacillus subtilis (NCIM 2250)

0.62

1.25

0.31

0.48

Pseudommonas aeruginosa (NCIM 2036)

1.25

0.31

0.31

0.60

Klebsiella pneumoniae (NCIM 2719)

0.62

0.62

0.15

0.72

Bacillus pumilis (NCIM 2327)

0.31

1.25

1.25

0.96

Candida albicans (NCIM 3471)

1.25

0.62

1.25

0.48

Aspergillus niger (NCIM 545)

2.5

1.25

0.15

0.24

Fusarium monoliforme (NCIM 1099)

1.25

1.25

0.62

0.24

Bacteria

Fungi

Trichoderma viridae (NCIM 1221)

0.62

0.62

0.31

0.48

Phanerochaete chrysosporium (NCIM 1197)

1.25

1.25

1.25

0.96

Pcilomyces species (NCIM 1081)

2.5

1.25

1.25

0.96

Values are inhibition zone (mm), and an average of triplicate.

Each extract has concentration of 500 g/ml,
Standard Drugs: Amoxycillin (10 g/disc), Ketoconazole (50 g/disc)
Incubation conditions for bacteria1 day at 370 C. For fungi7 days at 270 C.

34

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Apr-Jun 2011

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Jain Pritam et al: Antimicrobial properties of Abelmoschus manihot Linn

References:
1)

Mathew J. Antimicrobial activity of selected aromatic

plants. Ind Drugs 2005; 42: 28-32.

2)

The Anonymous, The Wealth of India: Raw

Materials, Vol. X. Publication and Information
Directorate, CSIR, New Delhi, India, 1976, pp. 588.

3)

Linlin WU, Xinbo Y, Zhengming H, Hezhi L.and

Guangxia WU. In vivo or in vitro antiviral activity of
hyperoside extracted from Abelmoschus manihot
medic. Acta Pharmacol Sin 2007; 28(3): 404-407.

4)

Puel C, Mathey J, Davicco MJ, Lebecque P,

Chanteranne

B,

Horcajada

MN,

Coxam

V.

Preventive effect of Abelmoschus manihot (L.)

Medik on bone loss in the ovarectomised rats. J
Ethnopharmacol 2007; 99: 55-57.
5)

Vaiiidi H, Kamalinejad M, Sedaghati S. Antimicrobial

properties of Croccus sativus L. Iranian J. Pharm
Res 2002; 1: 33-35.

6)

Wang RX, Zhao Q, Ying. Chih WU. Studies on the

chemical constituents of Abelmoschus manihot.
Zhongguo Zhong Yao Zhi 1981; 23(3): 222-225.

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