S114 Asian Pacific Journal of Tropical Disease (2012)S114-S117

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Asian Pacific Journal of Tropical Disease

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Document heading

Phytochemical screening and evaluation of anti-inflammatory activity of

methanolic extract of Abroma augusta Linn
Sutapa Das, Rana Datta, and Subhangkar Nandy
Department of Pharmacology, Gupta College of Technological Sciences, Ashram More, Asansol-713301, West Bengal, India

ARTICLE INFO ABSTRACT

Article history: Objective: To evaluate the Phytochemical and anti-inflammatory property of the different parts
Received 15 June 2012 of methanolic extracts of Abroma augusta Linn. Materials and methods: Abroma augusta
Received in revised form 27 June 2012 Linn (Family-Malvaceae) commonly known as Ulatkambal in Hindi and Devil’s cotton in
Accepted 18 October 2012
English, found in tropical Asia, South and eastern Africa, and Australia. It is mainly used for
Available online 28 October 2012
dysmenorrhoea, ammenorrhoea, sterlilty and other menstrual disorder. The present study aimed
at evaluation of Phytochemical and anti-inflammatory study of different parts of Abroma augusta
Keywords: Linn methanolic extract by the carrageenan induced rat paw oedema method. Results: The
Abroma augusta Linn (Family-Malvaceae) result showed significant anti-inflammatory property of different parts of Abroma augusta Linn
Antiinflammatory activity methanolic extract. Conclusions: The methanolic extract of different parts of Abroma augusta
Phytochemical study Linn showed potent activity comparing with the standard drug diclofenac sodium perhaps due to
Rat paw oedema the alkaloids and flavonoids present in the plant.

1. Introduction inflammatory disorders and related diseases have been

treated with plants or plant-derived formulations [4, 5].
I nflammation plays an important role in various The anti-inflammatory activity of several plant extracts
diseases, such as rheumatoid arthritis, atherosclerosis and isolated compounds has already been scientifically
and asthma, which all show a high prevalence globally. demonstrated.
During an inflammatory response, mediators, such as pro- Abroma augusta Linn (Family-Malvaceae) commonly
inflammatory cytokines, including interleukin IL-1, tumour known as Ulatkambal in Hindi and Devil’s cotton in English.
necrosis factor (TNF), interferon (INF)-c, IL-6, IL-12, IL- A genus of evergreen plant large, spreading, quick-growing
18 and the granulocyte-macrophage colony-stimulating hairy shrub or a small tree with velvety branches, found in
factor, are released; this response is antagonised by anti- tropical Asia, South and eastern Africa, and Australia. It
inflammatory cytokines, such as IL-4, IL-10, IL-13, IFN-a is mainly used for dysmenorrhoea, ammenorrhoea, wound
and the transforming growth factor. The nuclear factor-jB healing, sterlilty and other menstrual disorder. Powdered
(NF-jB), transcription factor, also plays an important role root act as anabortifacient and anti-fertility agent. Leaves
in the inflammatory response by regulating the expression are useful in treating uterine disorders, diabetes, rheumatic
of various genes encoding pro-inflammatory cytokines, pain of joints, and headache with sinusitis. Leaves and stem
adhesion molecules, chemokines, growth factors, and are demulcent and an infusion of fresh leaves and stem in
inducible enzymes such as cyclooxygenase- 2 (COX-2) cold water is very efficacious in gonorrhea. The root-bark is
[1, 2] inducible nitric oxide synthase ( i NOS ) and COX - 2 used as an emmenagogue and uterine tonic. [6, 7, 8]
both stimulate the production of large amounts of pro- H owever this plant has not been studied for anti-
inflammatory mediators. I n chronic inflammation, the inflammatory activity. This study was aimed at providing
negative regulatory mechanism appears to be dysfunctional. pharmacologic basis for its folkloric use in inflammation.
Although inflammation is primarily a protective response Based on this an attempt has been made to evaluate the
(against micro-organisms, toxins or allergens, for example), inflammatory potency of Abroma augusta Linn with their
inflammation that is chronic and uncontrolled becomes phytoconstituents.
detrimental to tissues [3].
S ince ancient times, in various cultures worldwide,
2. Materials and methods
*Corresponding author: Subhangkar Nandy Department of Pharmacology, Vedica
College of Pharmacy, RKDF Group, Bhopal, (M.P.) - India. 2.1. Collection of plant materials
E-mail: [email protected]
Mobile: 07415366404, 09932290182.
 Sutapa Das et al./Asian Pacific Journal of Tropical Disease (2012)S114-S117
S115

The leaves, bark, root of Abroma augusta was collected 2.3. Treatment of animals
from Siliguri, Raigang, West Bengal, India. A herbarium
sheet was prepared & it was sent to A . J . C . B INDIAN Healthy male and female rats (Wistar albino) of 4-8 weeks
BOTANIC GARDEN, Shibpur, Howrah and West Bengal, old were selected after physical and behavioural veterinary
India for authentication. The authentication no. of the study examination from Institutional Animal House of Gupta
plant is” CNH/111/2011/Tech.II/627”. The leaves, bark, root College of Technological Sciences. The weight range was
of Abroma augusta was collected and dried under shade. fall within依20% of the mean body for each sex at the time of
These dried materials were mechanically powdered, sheaved initiation of treatment. All experiments involving animals
using 80 meshes and stored in an airtight container. These complies with the ethical standards of animal handling and
powdered materials were used for further Phytochemical and approved by Institutional Animal ethics committee (955/A/06/
anti-inflammatory study (Figure 1 and 2). CPCSEA).
Sixty young adult male Wistar rats, weighting 120-150
g were obtained from the Institutional Animal House of
Gupta College of Technological Sciences. The rats were
housed in polyethylene cages in the Animal House. The rats
were housed in polyethylene cages, allowed one week of
acclimatization, and maintained on standard rat chow and
standard laboratory conditions throughout the experiment.

2.4. Phytochemical Screening

T he concentrated extracts were used for preliminary

screening of various phytoconstituents viz. carbohydrate,
amino acid, alkaloids, tannins and flavonoids were detected
by usual methods prescribed in standard tests. [9]

2.5. Acute toxicity test

Acute toxicity study was performed as per OECD guidelines

Figure 1: Picture showing the Abroma augusta plant. 423. [10] (Acute toxicity class method).

2.6. In-vivo anti-inflammatory activity

Male or female Sprague-Dawley rats with a body weight

between 120 -180 g were used. The animals were starved
overnight. The animals were fasted for 18 hours prior to the
experiment. Animals were divided into five groups of six
animals each and marked. Group I received 1% Tween-80
(1%, i.p.) and served as control. Group II received Diclofenac
sodium 5mg/kg b.w. i.p. and served as standard. Group III,
group IV,group V received the leaf, bark and root extract
respectively at dose of 250mg/kg b.w. i.p each. One hour
after the administration (as per the experimental protocol),
0.1ml of 1% carrageenan solution was injected beneath the
sub-plantar surface of the right hind paw of all animals.
For the assessment of the anti-inflammatory activity, the
volume of the paw was measured with the help of digital
plethysmometer at 0h and at 1h interval for a period of
three hours after the carrageenan treatment. The results are
tabulated by % of inhibition. [11]

2.7. Statistical analysis

Figure 2: Picture of digital Plethysmometer(ORCHID SCIENTIFICS,
PFM-01) A ll the values ware statistically analyzed by one-
way analysis of variance (ANOVA) followed by multiple
2.2. Preparation of extract comparison test. Comparison between control and drug
treated groups were considered to be significant P<0.01,
The air dried crushed leaves, bark and roots (1000g) were
P<0.001. All values are expressed as mean依SEM.
soaked for 12 hr in Methanol (3L) at room temperature. The
residue was extracted with hot Methanol under reflux 3
times (each 1500 ml) after vacuum filtration. All solvent was 3. Results
evaporated under vacuum and extract was then lyophilized,
3.1. Acute toxicity studies
to yield approximately 12% w/w/) of the residue, which was
stored at 20 曟 until use. The concentrate was suspended in
The extracts of Abroma augusta did not show any sign
5% w/v Tween 80 and given at dose 1ml/100gm body weight.
S116 Sutapa Das et al./Asian Pacific Journal of Tropical Disease (2012)S114-S117

of toxicity up to 2000 mg/kg body weight and hence it was 0.9

considered to be safe. 0.8

Inhibition ofinflammation (mL)

0.7

3.2. Phytochemical studies 0.6

0.5

F rom the P hytochemical study, it has evaluated the 0.4

presence of alkaloid, carbohydrate, flavonoid and tannin 0.3

0.2
in leaves and bark. B ut roots contain lower alkaloids,
0.1
carbohydrate and tannins. (Table 1) 0
Control (0mg/kg) Standard (10mg/kg) Bark (250mg/kg) Root (250mg/kg) Leaves (250mg/kg)

3.3. In-vivo anti-inflammatory study Dose mg/mL

Methanolic Extracts were evaluated for Anti-inflammatory Figure 4: This histrogram showing the inhibition of carrageennan
activity. I n C arrageenan induced paw oedema the indused paw oedema in rats. Histrogram is poltted between control,
intraperitoneally administration of leaves, bark, root doses of standard drug Diclofenac sodoum (100mg/kg), extracts of bark,
produced a significant anti-inflammatory activity in a dose-
dependent manner respectively in the rats. Bark extract has root, leaves (250mg/kg) in X axis and inhibition of paw oedema in Y
shown significant anti-inflammatory effect than root and axis. Data are shows inhibition of paw oedema significantly decrease
leaves. (Table 2)(Figure 3 and 4) by extract of Bark than Root and Leaves from Control. Data are Mean
S.E.M. indicates significant decrease in inflammation form control and
indicates highly significant decrease, P<0.01, P<0.001.
0.8

0.7 4. Discussion
0.6
Inflammation is a common phenomenon and it is a reaction
0.5
of living tissues towards injury. Steroidal anti-inflammatory
agents will lyse and possibly induce the redistribution of
Response (mL)

Control
0.4 Standard lymphocytes, which cause rapid and transient decrease
Bark
in peripheral blood lymphocyte counts to affect longer
0.3 Root
Leaves term response. Abroma augusta of the family Malvaceae
0.2 is a common plant of Asia, South and eastern Africa, and
Australia. Phytochemical evaluation of the various extracts
0.1
of Abroma augusta reveals the presence of flavonoids,
0 carbohydrate, tannins and alkaloids. Here anti-inflammatory
0 50 100 150 200 activity was performed based on the folk lore information
Time (min) using rat paw oedema method.
Carrageenan induced inflammation is a useful model for
Figure 3: This graphical representation shows the various response the estimation of anti-inflammatory effect. The development
on rat in carrageenan induced paw oedema in rat in 15min, 30 min,
of oedema in the paw of the rat after the injection of
60min, 120min, 180min after drug administration. Graph is plotted
Carrageenan is due to the release of histamine, serotonin,
between. Time (min) in X axis and Response (ml) in Y axis. Data are
prostaglandin and the like [12-13]. There is good evidence
shows onset of response time significantly increase by extract of Root
that the early or first phase of transient permeability is due
than Bark and Leaves from Control. P<0.01,P<0.001.
to the release of histamine and can thus be suppressed by
antihistamines. The mediation of the delayed or second
phase of exudation is more controversial and complex,

Table 1
Preliminary Phytochemical screening of the various extracts of the leaves of Abroma augusta
Solvent
Chemical Constituent
Test Methanolic root extract Methanolic bark extract Methanolic leaves extract
Alkaloid 1.Dragendroff’s Reagent - - -
2.Mayer’s Reagent + + +
3.Wagner’s Reagent + + +
4.Hager Reagent - - -
Amino acid Millon’s Test - - -
Carbohydrate 1.Molish Test + + +
2.Barfoed’s Test + + +
Flavonoid Sample + Lead acetate + + +
Tanin Ferric Chloride Test + + +
 Sutapa Das et al./Asian Pacific Journal of Tropical Disease (2012)S114-S117
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Table 2
Effect of Methanolic extract of Abroma augusta on carrageenan induced inflammation
Paw volume after Carrageenan injection
DOSE
GROUP 15min 30min 60min 120min 180min
mg/kg
EV(ml) EI% EV(ml) EI% EV(ml) EI% EV(ml) EI% EV(ml) EI%
Control - 0.58依 0.0011 0.60依 0.0008 0.65依 0.0006 0.71依 0.0001 0.75依 0.0002
Standard 100 0.56依 0.0006 3.97 0.53依 0.0095 11.11 0.43依 0.0009** 34.5 0.039依 0.0010** 45.11 0.36依 0.0015** 51.33
Bark 250 0.56依0.0005 3.97 0.58依 0.0007 4.58 0.54依 0.0010* 14.20 0.50依 0.0005* 29.30 0.40依 0.0008** 46.90
Root 250 0.57依 0.0007 1.7 0.54依0.0010 7.14 0.55依 0.0002* 16.9 0.53依 0.0009* 26.03 0.51依 0.0009* 31.42
Leaves 250 0.58依 0.0003 5.67 0.55依 0.0002 6.66 0.56依 0.0007* 15.73 0.50依 0.0001* 30.22 0.48依 0.0001* 35.40
The data are expressed as mean依S.E.M. Significant differences in each group versus the control were as follows: * P < 0.05. ** P < 0.01.

and has been attributed in part to kinins, prostaglandins, [4] Krishnaswamy, K. Traditional Indian spices and their health
neutrophils, and lipoxygenase products of arachidonic acid significance. Asia Pacific Journal of Clinical Nutrition 2008; 17:
metabolism. The probable mechanism of anti-inflammatory 265-68.

action of Extract may be due to its influence on the second [5] Marc, E. B., Nelly, A., Annick, D. D., & Frederic, D. Plants used
phase of inflammation, the cyclooxygenase pathway rather as remedies antirheumatic and antineuralgic in the traditional
medicine of Lebanon. Journal of Ethnopharmacology 2008; 120:
than the lipoxygenase pathway. This is evident by the
315-34.
maximal inhibition of inflammation at the end of the third
[6] Babita gupta et al. Der Pharmacia Sinica, Pelagia Research
hour after the challenge with carrageenan[14, 15].
Library 2011; 2: 253-61.
Different parts of methanolic extracts of Abroma augusta
[7] H. Sh, A. Hanif, B. Agarwal, R. Mohammed and J. Rowank. A
showed significant anti-inflammatory activity. T his journal of plants, people and applied Research Ethnobotany
significant anti-inflammatory effect may be due to the research and applications. 2010; 8: 61-74.
inhibition of any inflammatory mediators by the alkaloids [8] M . M ishra, G . G hosh, D . M ishra , B . B arik. Advance in
and Flavonoids [16] present in the extract. The present result pharmacology and toxicology 2008; 9: 147-50
indicates the efficacy of Abroma augusta as an effective [9] Bodhisattwa Chakraborty, Subhangkar Nandy and Rana Datta.
therapeutic agent in the treatment of acute inflammations Phytochemical and Pharmacognostical Studies of the Leaves Of
[17]. The result of present study authentifies the folk lore Alangium Lamarkii. IJBR 2012; 3: 196-200.
information on the anti-inflammatory property of the leaf [10]TK Mohamed Saleem, AK Azeem, C Dilip, C Sankar, NV Prasanth,
extract of Abroma augusta. Further and detailed studies are R Duraisami. Anti-inflammatory activity of the leaf extacts of
in process for the isolation of active constituent responsible Gendarussa vulgaris Nees. Asian Pacific Journal of Tropical
for this property and to idendification of the possible Biomedicine 2011; 1: 147-49.
mechanism of its anti inflammatory property. [11]Jude E Okokon1, Anwanga E Udoh, Samuel G Frank, Louis U
Amazu. Anti-inflammatory and analgesic activities of Melanthera
scandens. Asian Pacific Journal of Tropical Biomedicine 2012; 1:
144-48.
Conflict of interest statement
[12]G e o r g e w i l l O A , G e o r g e w i l l U O , N w a n k w o a l a R N P .
Antiinflammatory effects of Morninga oleifera lam extract in rats.
We declare that we have no conflict of interest.
Asian Pac J Trop Med 2010; 3: 133-35.
[13]G eorgewill OA , G eorgewill UO . E valuation of the anti-
inflammatory activity of extract of Vernonia Amygdalina. Asian
Acknowledgements Pac J Trop Med 2010; 3: 150-51.
[14]R osa MD , G iround JP , W illoghby DA . S tudies of the acute
T he authors are thankful to P rof. D ebesh C handra inflammatory response induced in rats in different sites
Mazumder (Chairman, Trinity Trust,Asansol,W.B), Dr. Kalyan byCarrageenan and turpentine. J Pathol 1971; 104: 15-29.
Kumar Sen (Principal, G.C.T.S., Asansol, W.B.). [15]Sheetal S. Chaudhari, Sanjay R. Chaudhari, Machindra J. Chavan.
Analgesic, anti-inflammatory and anti-arthritic activity of Cassia
uniflora Mill. Asian Pacific Journal of Tropical Biomedicine 2012;
Reference 2: 181-86.
[16]Haroon Khan, Murad Ali Khan, Naveed Muhammad, Nadeem
[1] Hanada, T., & Yoshimura, A. Regulation of cytokine signaling and A shraf, F arah G ul, S hafiq A hmad T ariq. A nti-inflammatory
inflammation. Cytokine and Growth Factor Reviews 2002; 13: 413- and antioxidant activity of Joshanda partially mediated through
21. inhibition of lipoxygenase. Phytopharmacology 2012; 3: 19-28.
[2] M akarov, S . S . NF -kb as a therapeutic target in chronic [17]Khan I, Nisar M, Ebad F, Nadeem S, Saeed M, Khan H, Samiullah,
inflammation: Recent advances. Molecular Medicine Today 2000; Khuda F, Karim N, . Ahmad Z.. Anti-inflammatory activities
6: 441-48. of Sieboldogenin from Smilax china Linn.: experimental and
[3] Gil, Ã. Polyunsaturated fatty acids and inflammatory diseases. computational studies. Journal of Ethnopharmacology 2009; 121:
Biomedicine and Pharmacotherapy 2002; 56: 388-96. 175-77.