WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Salvi et al. World Journal of Pharmacy and Pharmaceutical Sciences

Volume 2, Issue 6, 5199-5205. Research Article ISSN 2278 – 4357

EVALUATION OF ANTI-INFLAMMATORY ACTIVITY OF AN

ETHANOLIC EXTRACT OF THE SEEDS OF ENSETE SUPERBUM
ROXB.

Vidula Salvi*, Siddhesh Mungekar, Vaibhavi Garge, Yadunath Joshi, Vilasrao Kadam

Bharati Vidyapeeth’s College of Pharmacy, Sector-8, C.B.D. Belapur, Navi-Mumbai- 400614.

Article Received on ABSTRACT

25 August 2013, The intention of the present study was to evaluate the anti-
Revised on 21 Sept 2013, inflammatory activity of an ethanolic extract of seeds of the plant
Accepted on 11 November
2013 Ensete superbum Roxb. (E.S.); belonging to the family Musaceae.
Traditionally, Ensete superbum is used in psychotic disorders, diabetes
*Correspondence for mellitus, stomach pain, dysuria and leucorrhoea. On phytochemical
Author: evaluation it was found that the plant contains glycosides, flavonoids,
alkaloids, carbohydrates, phenols and tannins. As per World Health
* Vidula Salvi
Bharati Vidyapeeth’s College Organisation (WHO) guidelines for Quality Standardised Herbs and
of Pharmacy, Sector-8, C.B.D. Herbal Formulation; determination of various physico-chemical
Belapur, Navi-Mumbai- parameters like foreign matter, moisture content, total ash, water
400614, India.
soluble ash, acid insoluble ash, alcohol soluble extractive value and
vidu.salvi1411@gmail.com
water soluble extractive value were performed. The extraction was
carried out with Soxhlet apparatus, using ethanol. The animals used for
study were female Sprague Dawley rats with 120-140 grams of body weight and further
divided in four groups of six animals each. Group I was of control (vehicle), Group II of (E.S.
250 mg/kg), Group III of (E.S. 500 mg/ kg) and Group IV of standard (Indomethacin
10mg/kg). The investigation of Ensete superbum as anti-inflammatory drug was performed
using phlogistic agent (Carrageenean) induced paw edema. 0.1 ml of 0.1% of carrageenean
was injected in sub-plantar region of right paw of the rat and paw volume was measured at a
specific time interval after treatment. At both the doses of an ethanolic extract of the seeds of
Ensete superbum showed a significant anti-inflammatory activity when compared to the
standard and control groups.

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Keywords: Ensete superbum, anti-inflammatory, carageenean, ethanolic.

1. INTRODUCTION
During the period of 2500 to 500 BC, the concept of Ayurveda appeared and developed in
India. The literal meaning of Ayurveda is “Science of life” because ancient Indian system of
health care focused views in man and illness.[1] Charak Samhita (900 BC) is the first recorded
treatise of Ayurveda and describes 341 plant and plant products for use in medicine. The next
landmark in Ayurveda is Sushrata Samhiti (600 BC) with special emphasis on surgery and
this treatise 395 medicinal plants.[2] India, due to its unique variety of geographical and
climatic factors, has had a rich and varied flora of medicinal plants since the Vedic period.
No wonder that in India plants in form of crude plants have been used since ancient time for
the treatment of human ailments. With advent of science, synthetic drugs became more
popular and replaced the traditional systems of medicine. Though, the modern medicines are
more efficient in curing diseases, they produce number of side effects, also these drugs many
a times are incapable of preventing progress of disease.[3] On the contrary, the crude drug are
less efficient with respect to cure disease but they are relatively free from side effects which
have made the traditional medicine survive the competition of modern day medicine. Also for
inflammation, there are synthetic non steroidal anti-inflammatory drugs; but prolonged use of
these drugs for inflammatory disease like rheumatoid arthritis there occur side effects like;
epigastric pain, gastro-intestinal bleeding and ulcers. These drugs suppress the inflammation
and providing only symptomatic relief. Hence, the search for new anti-inflammatory drugs
that retain therapeutic efficacy and yet which are devoid of these side effects.[4]

Ensete superbum Roxb.(wild banana or raan keli) belongs to family Musaceae and it is 12
feet tall evergreen perennial shrub. Plants may grow up to 12 ft in height and the pseudostem
may be up to half the height with a swollen base of up to 8 ft in circumference at the base.
The leaves are bright green in colour on both sides with a deeply grooved and short petiole.
The leaf sheaths are persistent at the base and leave closely set scars on the corm. The fruits
are about 3 inches long and more or less triangular with dark brown seeds. The upper parts of
the plant die out during the dry season leaving the corm, which forms new leaves at the
beginning of the monsoon.[5] It is widely distributed in the regions of Western ghats,
Anamalai hills and hills in Dindigul and Jhadol and Ogna forest ranges in Rajasthan. Ensete
superbum has been traditionally used in psychotic disorders, kidney disorders, dog bite,
diabetes, dysuria, stomach pain and anti-oxidant. [6]

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2. MATERIALS AND METHOD

2.1. Collection of plant materials
The seeds of Ensete superbum Roxb. were collected from a local Ayurvedic medicinal shop,
Kalbadevi, Mumbai-400002, India and were authenticated by Dr. Harshad M. Pandit (Head
of Department and Associate Professor of Botany), Guru Nanak Khalsa College, Matunga,
Mumbai-400019, Maharashtra. The voucher specimen (vvs/130912b) has been preserved for
further reference.

2.2. Preparation of extracts

The seeds of Ensete superbum were shade dried and were further ground to coarse powder.
The powdered drug was subjected to extraction using ethanol as solvent. Soxhlet apparatus
was used for the purpose of extraction. The solvent was completely removed under reduced
pressure and semisolid mass was obtained and it was stored in a vacuum dessicator for
further use. Suspension of two different doses of 250mg/kg and 500mg/kg according to the
body weight were prepared, tween-80 was used as a suspending agent.

2.3. Animals used

For the study purpose, female Sprague Dawley rats (120-140 grams) were used. The animals
were procured from Glenmark Pharmaceuticals, Ghansoli, Navi-Mumbai, India; and were
housed in animal house of Bharati Vidyapeeth’s College Of Pharmacy, Belapur, Navi-
Mumbai, India. They were caged in polyvinyl cages and feed with standard food pellets and
received water ad libitum. All the animals were maintained in a group of six at 22to 25oc
temperature with the light and dark cycle of 12:12 hours. The animals were starved overnight
for 16 hrs before experiment.

2.4. Preliminary Phytochemical screening [7]

For the presence of plant phyto constituents such as carbohydrates, alkaloids, glycosides,
flavonoids, tannins and saponins preliminary phytochemical screening were carried out.

2.5. Physico-chemical parameters [8]

The determination of various physic-chemical parameters such as foreign matter, moisture
content, total ash value, water soluble ash, acid insoluble as, alcohol soluble extractive value
and water soluble extractive value were performed as per WHO guidelines for Quality
Standardized Herbs and Herbal Formulation.

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2.6. In-vivo Anti-inflammatory activity using Carrageenean induced rat paw edema
model [9]
Ensete superbum was evaluated for anti-inflammatory activity with the help of Carrageenean
induced rat paw edema method. Female Sprague dawley rats were used randomly separated
randomly in 4 groups of 6 animals in each. Group I was of control (vehicle), Group II of
(Indomethacin 10mg/kg of body weight of animal), Group III of (E.S. 250 mg/kg of body
weight of animal), and Group IV of (E.S. 500 mg/ kg of body weight of animal). 30 minutes
before sterile injection of the phlogistic challenge, the animals were given the doses of
vehicle, test (extracts) and standard (Indomethacin) according to the groups. After oral
administration of dose of vehicle, extracts and standard, the rats were challenged by a
subcutaneous injection of Carrageenean (0.1 ml of 1 % w/v suspension in 0.9% saline)
subplantar side of the left paw. Inflammation was measured in terms of ml i.e. displacement
of water by edema using a plethysmometer immediately after carrageenean injection. Paw
volume was measured using water displacement plethysmometer at the time interval of 0, ½,
1, 2, 3, 4, 5, 6 hrs.

The percentage inhibition was calculated by using the formula percent inhibition:

Vc-Vt/Vc X 100

Vc= edema volume of control

Vt= edema volume of test.

3.7 Statistical analysis

The data were analyzed using one way ANOVA statistical methods. For the statistical tests a
P values of less than 0.001, 0.01 and 0.05 was taken as significant.

3. RESULTS AND DISCUSSION

3.1. Preliminary phytochemical screening
The preliminary phytochemical screening revealed the presence of glycosides, flavonoids,
alkaloids, carbohydrates, phenols and tannins in the ethanolic extract of seeds of Ensete
superbum Roxb.

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3.2. Physico-chemical parameters

As per WHO guidelines for Quality Standardized Herbs and Herbal Formulation physico-
chemical parameters for Ensete superbum Roxb. revealed the following results:
Table no. 1

Physic-chemical parameters Value in percent (%)

Foreign matter 0.16
Moisture content 0.9
Total ash value 3.2
Water soluble ash value 1.54
Acid insoluble ash value 0.61
Alcohol soluble extractive value 6.7
Water soluble extractive value 24.93

3.3. In-vivo Anti-inflammatory activity using Carrageenean induced rat paw edema
model
Table no. 2
Ethanolic extract
Ethanolic extract of
Standard of Ensete
Control Ensete Superbum
Time (Indomethacin Superbum
Parameter (vehicle) Roxb (EESS)
(hrs) 10mg/kg body Roxb.(EESS)
(500 mg/kg body wt.
wt. of animal) (250 mg/kg body
of animal)
wt. of animal)
0.27±0.0 0.24±0.03 0.26±0.02 0.25±0.02
0.5
3 (11%) (3.7%) (7.4%)
0.34±0.0 0.25±0.04 0.29±0.02 0.27±0.02
1
4 (24.47%) (14.70%) (20.58%)
0.55±0.0 0.34±0.01 0.44±0.02 0.42±0.02
2
5 (38.18%) (23.63%) (23.63%)
0.75±0.0 0.31±0.02** 0.34±0.03* 0.32±0.02**
3
2 (58.04%) (54.44%) (57.33%)
0.62±0.0 0.29±0.04 0.33±0.04 0.30±0.03
Edema 4
7 (52.70%) (46.77%) (51.61%)
Volume
(ml) 0.51±0.0 0.26±0.05 0.29±0.05 0.27±0.03
5
2 (48.00%) (43.13%) (47.05%)
0.45±0.0 0.29±0.04 0.32±0.02 0.31±0.02
6
3 (35.55%) (28.88%) (31.11%)

All values are expressed as mean ± SEM (n=6) and *p≤0.05 less significant, ** p≤0.01
significant, *** p≤0.001 highly significant as compared with control and standard.

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Graph no. 1

Carrageenean induced hind paw edema is the standard experimental model of acute
inflammation. Carrageenean is the phlogistic agent of choice for testing anti-inflammatory
drugs as it is not known to be antigenic and is devoid of apparent systemic effects. Moreover,
[9]
the experimental model exhibits a high degree of reproducibility . Carrageenean induced
edema is a biphasic response. The first Phase is mediated through the release of histamine,
serotonin and kinins whereas the second phase is related to the release of prostaglandin and
slow reacting substances which peek at 3h[10]. It has been reported that the second phase of
edema is sensitive to drugs like hydrocortisone, phenylbutazone and indomethacin.

The present study of anti-inflammatory activity of ethanol extract of Ensete superbum Roxb.
against carrageenean induced paw edema shows that the extracts have significant effect on
inflammation and markedly reduced the swelling. The percentage reduction in the paw
volume in the group of animals treated with ethanolic extract of Ensete superbum Roxb
250mg/kg of body wt. of animal was 54.44% and for the 500mg/kg of body wt. of animal
was 57.33% at 3 hours. It shows that the seeds extract of the plant Ensete superbum Roxb
have significant (* p <0.05, ** p < 0.01, *** p < 0.001) anti-inflammatory effect and the results
were compared with Indomethacin 10mg/kg of body weight of animal and shown a
percentage paw volume reduction of 58.04 %. (Table no.2)

CONCLUSION
The ethanolic extract of seeds of Ensete superbum Roxb. was screened for the in-vivo anti-
inflammatory activity using carrageenean induced rat paw edema model. These activity may
be due the presence of glycosides, flavonoids, alkaloids. At the dose of 500mg/kg of extract,

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Salvi et al. World Journal of Pharmacy and Pharmaceutical Sciences

showed a better anti-inflammatory activity. This study demonstrates the efficacy of seeds of
Ensete superbum as anti-inflammatory agent and also scientifically justifies the use of this
plant as an anti-edematous agent in folk medicine. However, further study is required to
determine the actual constituents responsible for anti-inflammatory activity and further
authenticate its mechanism of action.

ACKNOWLEDGEMENT
We are thankful to Dr. Vilasrao Kadam Principal of Bharati Vidyapeeth’s College Of
Pharmacy for providing us with necessary facilities.

REFERENCES
1. Samy RP, Pushparaj PN, Gopalkrishnakone P. A Compilation Of Bio-Active Compounds
From Ayurveda. Bio-information (2008). 3(3); 100-110.
2. Patel SS, Saluja AK. Traditional Medicine Sources Of New Drugs. Pharma Times 34,
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3. Robber JE, Tyler VE. Tyler’s Herb of Choice: The Therapeutic Use of Phytomedicinals
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4. Singh S, Majumdar DK. Ocimum sanctum- Phyto Chemical and Pharmacological
Evalution. Recent progress of medicinal plants 2, Page no. 1-81.
5. Constantine, D. (1999) The Musaceae — an annotated list of the species of Ensete, Musa
and Musella.
6. Sharma, S.K. (1996). “Presence of Wild Plantain (Ensete superbum) in Rajasthan”.
Journal of the Bombay Natural History Society 93(2): 322-323.
7. Kokate CP, Purohit AP, Gokhale SB. (2007), Pharmacognosy, Nirali Prakashan.; 39th
edition: 607-611.
8. Dr. Khandelwal KR. Practical Pharmacognosy Techniques and Experiments. Nirali
Prakashan; Content-23; Pageno-23.1-23.7.
9. Winter CA, Risley E, Nuss G. Carrageenean Induced Edema In Hind Paw Of The Rats As
An Assay For Anti-Inflammatory Drugs. Proc. Soc Exp Biol Med. 1962; 111: 544-547.
10. Vinegar R, Schreiber W, Hugo R ,1969. Biphasic Development of Carrageenan Edema on
Rats. J. Pharmacol. EXP. Ther. 66:96-10.

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