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Pharmacologyonline 2: 290-298 (2006) Tangpu and Yadav
290
ANTIDIARRHOEAL ACTIVITY OF CYMBOPOGON CITRATUS
AND ITS MAIN CONSTITUENT, CITRAL
Vareishang Tangpu and Arun K. Yadav*
Department of Zoology, North-Eastern Hill University, Shillong 793 022, India.
*Corresponding Author: akynehu@hotmail.com
Summary
Cymbopogon citratus (DC) Stapf is one of the most commonly used plants in the
folk medicine of native tribes of northeast India to cure diarrhoeal disorders. Hence
the present study was carried out to verify the antidiarrhoeal claims of C. citratus
extract and its main constituent, citral using three experimental diarrhoeal models
in mice: castor oil induced-diarrhoea, MgSO4 - induced enteropooling and charcoal
meal test. The effects of extract and citral were also observed on faecal out put in
mice. The 800 mg/kg, p.o. dose of plant extract significantly reduced the
production of faecal matters by 53.44%, and conferred protection to diarrhoeal
episodes after castor-oil challenge by 59.00%. The same dose of extract also
significantly inhibited the intestinal fluid secretion induced by MgSO4 and
gastrointestinal motility in charcoal meal test. In all the experimental models, the
citral showed an almost comparable efficacy with that of standard antidiarrhoeal
drug, Loperamide. The study thus authenticates the presence of antidiarrhoeal
activity in Cymbopogon citratus, which may have therapeutic benefits in humans
encountering diarrhoeal disorders.
Keywords: Antidiarrhoeal activity, Cymbopogon citratus, citral, folk medicine,
India.
The rich floral diversity of India has provided traditional practitioners in the
country with an impressive pool of ‘natural pharmacy’ from which plants are
selected as ingredients to prepare herbal remedies to treat various diseases. The
northeastern region of India, in particular is inhabited by approximately 130 major
tribal communities. These tribes have a good faith in their traditional medicine
system and thus they use many plant based medicines to cure various ailments,
including diarrhoea. However, the purported efficacies of many of these plants
have not been scientifically evaluated. During course of our studies on
experimental validation of folk medicines of northeast India [1, 2, 3, 4], we
collected information through a random survey in the region that Cymbopogon
citratus (DC) Stapf. (Poaceae: Gramineae), locally known as Harvosing, is one of
the most commonly used herbal plants to cure diarrhoeal disorders. In the literature
there is neither any mention nor any experimental study pertaining to its
antidiarrhoeal activity, though other studies on this plant indicate that it possesses
scientifically proven antibacterial [5, 6, 7], antifungal [8, 9, 10, 11], antimalarial
[12], anticancer [13] and larvicidal activities against Aedes aegypti [14].
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C. citratus (DC) Stapf. is a perennial grass that grows up to 1.5 meters high. It is found distributed
in tropical Asia, including Penisula of India and Naga Hills in northeast India [15]. The main
chemical constituent of the plant is citral, which accounts for 75 to 80% of its oil’s volume [8, 16].
In practice, the whole stalk and the leaf are boiled and the decoction is drunk to relieve the
diarrhoea. In view of its popular use in traditional medicine system, it was felt necessary to
scientifically evaluate the antidiarrhoeal potentials of C. citratus as claimed by indigenous tribes in
India. The present study reports the antidiarrhoeal efficacy of C. citratus stalk decoction and its
main chemical constituent citral, using experimental diarrhoeal models in mice.
Methods
Plant material and preparation of extracts:
The plant material was collected in the month of August from surroundings of cultivated lands in
Manipur by V. Tangpu and later duly identified by Dr. P. B. Gurung, Department of Botany,
NEHU, Shillong. The leaves and stalk were air-dried under shade and pulverized into husky
powder. The powdered material was extracted by maceration in 2 L of cold distilled water for 24
h. The material was filtered and freeze dried to obtain the extract as a solid residue. The w/w yield
in terms of drying starting material was 10.50%.
Drugs and chemicals:
Loperamide (Axar Pharmaceuticals, Baroda), Castor oil (S. D. Fine, Mumbai), Activated Charcoal
(E. Merck, India), Gum Acacia (S. D. Fine Chem, Boisar) were used in this study.
Experimental Animals:
Six to eight weeks old mice (20-30 g) were used. The animals were acclimatized for 15 days under
the standard laboratory conditions following procurement from Pasteur’s Institute, Shillong. All
the animal experiments were carried out in accordance with the Rules and Regulations approved
by the Institutional Animal Care and Use Committee.
Preliminary acute toxicity test:
The C. citratus stalk extract was administered orally in the doses of 100, 200, 400, 800, 1600 and
3200 mg/kg, p.o. to six animals in each group. The animals were observed for mortality, if any,
and adverse signs in terms of body weight, body temperature, and food and water intake for 72 h
post administration of extract.
Antidiarrhoeal Activity:
A. Measurement of faecal output: Six groups of mice (n = 6) were housed singly in separate
cages. Group I served as the control and received 2% gum acacia (0.5 ml); Groups II – IV mice
were treated with 200, 400 and 800 mg/kg of plant extract. Group V mice received citral while,
Group VI mice were given 0.5 ml of 5 mg/kg Loperamide HCl, the standard antidiarrhoeal drug.
The faecal materials collected for 12 h post treatment, were dried in an incubator and their weights
measured. The faecal output of animals were calculated and expressed in terms of percentage
reduction [17].
B. Castor oil model:
Overnight-fasted mice were randomly divided into six groups (n = 6). Group I received 0.5 ml of
2% gum acacia suspension; groups II – IV were treated with 200, 400 and 800 mg/kg of plant
Pharmacologyonline 2: 290-298 (2006) Tangpu and Yadav
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extract; Group V mice were given 5 mg/ kg of citral while group VI received 0.5 ml of 5 mg/kg of
Loperamide. 1 h later, diarrhoea was induced in all groups by inoculating castor oil (0.5 ml/mouse,
p.o.). The numbers of diarrhoeal episodes were recorded for each time and cumulative values were
calculated for 4 h post induction of diarrhoea [18, 19].
C. Enteropooling assay:
Overnight-fasted mice were randomized into seven groups (n = 6). The animals received a
diarrhoeal agent (0.5 ml of 10% MgSO4/per mouse p.o.). Group I served as the control (0.5 ml; 2%
gum acacia); Group II served as a vehicle control (10% MgSO4 + 2% gum acacia); groups III – V
received 200, 400 and 800 mg/kg of plant extract, respectively; group VI received 5 mg/kg dose of
citral and group VII animals were given 5 mg/kg Loperamide. All these treatments were done 1 h
prior to diarrhoeal induction. 30 min later, animals were sacrificed and their small intestines were
ligated from pyloric sphincter to ileocaecal junction, and assessments of the accumulation of
intestinal fluid secretion induced by MgSO4 were made and expressed as percentage reduction in
fluid secretion [20].
D. Gastrointestinal transit test:
The animals were starved for 16 h prior to the experiment. The test extract (200, 400 and 800
mg/kg) was given orally to groups II – IV of mice (n = 6). Group I served as the control, group V
animals received 5 mg/kg of citral, while group VI was given 5 mg/kg Loperamide. 5 min later,
0.5 ml of charcoal meal was orally inoculated to each mouse. All the mice were sacrificed 30 min
later, their small intestines from pylorus to caecum cut out and distance travelled by the charcoal
marker measured, and expressed as a percentage of the total length of small intestines. The
percentage inhibition of the marker transit in the intestine was calculated as described by Akah &
Offiah [21].
Statistical analysis:
The results are expressed as mean ± standard error of mean (SEM). Significance of the result was
analyzed using Student’s t-test. P values < 0.05 were considered as significant.
Results
Acute toxicity test:
The plant extract administered orally to the mice up to 3200 mg/kg, p.o. did not show any
mortality or any adverse signs in the animals in terms of body weight, body temperature, and food
and water in take during 72 h period of observation.
Antidiarrhoeal activity:
Effect on faecal output:
The 800 mg/kg dose of extract reduced the faecal output by 53.44%. The reduction in faecal
output by Loperamide and citral was 57.01 and 45.37%, respectively. The results (Table 1) were
significantly different from the control value at P < 0.05 (200 mg/kg) and at P < 0.001 (400, 800
mg/kg extract and 5 mg/kg Loperamide HCl).
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Table 1 Effect of Cymbopogon citratus extract and citral on faecal output in mice
Treatment Wt. (g) of the dried faecal % %
(mg/kg, p.o.) matter per 100 g mouse* Faecal output Reduction in
faecal output
Control 0.842 ± 0.058 100.00 0.00
(2% gum acacia)
Plant extract
200 0.686 ± 0.021
a 81.47 18.53
400 0.445 ± 0.037
b 52.85 47.15
800 0.392 ± 0.007
b 46.56 53.44
Citral 5 0.460 ± 0.010
b 54.63 45.37
Loperamide
5 0.362 ± 0.021
b 42.99 57.01
* Data represent mean ± SEM from six animals. aP < 0.05, and bP < 0.001 as compared with
control group.
Effect on castor oil-induced diarrhoea:
The number of diarrhoeal episodes at each time in extract treated mice showed significant
difference from the control value. The onset of diarrhoeal droppings was significantly delayed in
the treated mice following castor oil challenge, and also the numbers of diarrhoeal episodes
(cumulative value) for 4 h were significantly inhibited in the treated groups (Table 2). The
percentage protection of diarrhoea at the end of 4 h was 50.00% by 800 mg kg dose of plant
extract, and 5 mg/kg doses of citral and loperamide, respectively.
Table 2 Effect of Cymbopogon citratus extract and citral on castor oil induced diarrhoea in
mice
Treatment No. of diarrhoeal episodes at time* %
at – 60 h
(mg/kg, p.o.) 1 h 2 h 3 h 4 h Protection
Control 4.33 ± 0.56 6.67 ± 0.71 8.33 ± 0.49 8.67 ± 0.42 0.00
(2% gum acacia)
Plant extract
200 0.83 ± 0.65a 1.50 ± 0.72b 2.00 ± 0.89b 2.17 ± 0.83b 33.33
400 0.67 ± 0.49b 1.33 ± 0.71b 1.83 ± 0.70b 1.83 ± 0.70b 33.33
800 0 b 0.50 ± 0.34b 1.17 ± 0.65b 1.50 ± 0.96b 50.00
Citral 5 0
b 0 b 1.17 ± 0.54b 1.17 ± 0.54b 50.00
Loperamide
5 0b 0.3 3± 0.21b 0.50 ± 0.22b 0.50 ± 0.22b 50.00
* Data represent mean ± SEM from six animals.aP < 0.01, and bP < 0.001 as compared with
control group.
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Effect on enteropooling assay:
The extract reduced significantly the diarrhoeal fluid accumulation, evoked by induction of
MgSO4, both in terms of small intestinal weights and measure of the accumulated fluids in small
intestines. 800 mg/kg dose of extract yielded 22.82% reduction. Whereas the reduction was
observed to be 26.51% by Loperamide. The citral (5 mg/kg) showed the maximum reduction of
27.85% (Table 3).
Table 3 Effect of Cymbopogon citratus extract and citral on MgSO4 - induced enteropooling
in mice
Treatment Vol. (in ml) of the small intestinal %
(mg/kg, p.o.) fluids accumulated per 100 g mouse* Reduction
Normal Control 1.40 ± 0.16 -
(0.5 ml saline)
Vehicle Control 2.98 ± 0.17 0.00
(0.5 ml saline+ MgSO4
Plant extract
200 2.55 ± 0.07
a 14.43
400 2.34 ± 0.15
b 21.48
800 2.30 ± 0.09
c 22.82
Citral 5 2.15 ± 0.10
c 27.85
Loperamide 5 2.19 ± 0.16 c 26.51
* Data represent mean ± S.E.M. from six animals. aP < 0.05, bP < 0.02, and cP < 0.01 as compared
with vehicle control group.
Effect on charcoal induced gastrointestinal transit:
The extract showed a dose-dependent inhibition of the charcoal marker in the small intestine of
treated animals. The motility inhibition by 57.22% in 800 mg/kg extract treated animals was
slightly higher to the inhibition observed (55.83%) for Loperamide. The values of the distance
travelled by charcoal marker are significantly (P < 0.001) different from the control value (Table
4).
Table 4 Effect of Cymbopogon citratus extract and citral on gastrointestinal transit in mice
Treatment Distance (cm) traveled by charcoal %
(mg/kg, p.o.) marker as % of the total length of Inhibition
small intestine *
Control 81.66 ± 0.54 0.00
(2% gum acacia)
Plant extract
200 69.26 ± 1.40
a 15.18
400 53.29 ± 1.82
a 34.74
800 34.93 ± 0.43
a 57.22
Citral 5 42.74 ± 2.84
a 47.66
Loperamide 5 36.07 ± 1.71 a 55.83
Data represent mean ± SEM from 6 animals. aP < 0.001 as compared with control group.
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Discussion
This study was aimed at evaluating the antidiarrhoeal potentials of C. citratus stalk decoction,
which is widely used in the treatment of diarrhoeal diseases by indigenous tribes of northeast
India.
The main feature of the small intestine is to absorb and secrete. Diarrhoea results from an
imbalance between the absorptive and secretive mechanisms in the intestinal tract, accompanied
by intestinal hurry, which results in an excess loss of fluid through the faeces [22]. Many animal-
based studies have investigated the bioactivity and effects on intestinal function of plants
traditionally used as treatment for diarrhoea [2, 17, 19]. Plant extracts can have antispasmodic
effects, delay gastrointestinal transit, suppress gut motility, stimulate water adsorbption or reduce
electrolyte secretion. These activities may explain the benefits of using a particular plants in the
treatment of diarrhoeal diseases [23]. Some of these experimental models were therefore employed
to evaluate the antidiarrhoeal potentials of C. citratus stalk extract in the present study.
The results demonstrated that the plant extract reduced the faecal output as compared to the control
group in a dose-dependent manner. This indicates towards the presence of an antisecretory or
proabsorbtive property in the extract. A significant inhibition in the diarrhoeal droppings after
castor oil induction observed in the present study, further suggests that the test extract has the
property to inhibit both secretory and motility mechanisms of diarrhoea. These results which
showed ability of plant extract to suppress the production and accumulation of wet faeces and an
inhibitory effect on gastrointestinal motility met the standard criterion to prove its efficacy as an
antidiarrhoeal agent. Castor oil is metabolized into ricinoleic acid, which in turn irritates and
causes inflammation in the intestinal mucosa, resulting in to release of prostaglandins. The
prostaglandins thus released stimulate the motility and secretion in the small intestines [23]. The
therapeutic effect of Loperamide is believed to be due to its antimotility and antisecretory
properties [24]. A significant inhibition in the diarrhoeal droppings after castor oil induction
observed in the present study, further suggests that the test extract has the property to inhibit both
secretory and motility mechanisms of diarrhoea.
Reduction of gastrointestinal motility is one of the mechanisms by which many antidiarrhoeal
agents can act [24]. It was observed that the extract suppresses the propulsion of charcoal marker
in a dose-dependent manner. In the present study the percentage inhibition of charcoal marker by
800 mg/kg dose of extract was observed to be slightly higher when compared to Loperamide. This
finding suggests that C. citratus extract has the ability to influence the peristaltic movement of
intestine indicating thereby the presence of an intestinal antimotility activity in it. These results
which showed ability of plant extract to suppress the production and accumulation of wet faeces
and an inhibitory effect on gastrointestinal motility met the standard criterion to prove its efficacy
as an antidiarrhoeal agent.
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In the small intestinal transit test the study showed that the plant extract suppress the propulsion of
charcoal marker in a dose-dependent manner. This finding suggests that the extracts appear to act
on all parts of intestine. The percentage propulsion of charcoal marker by C. citratus extract was
observed to be almost similar as that of Loperamide. The mode of action of loperamide is believed
due to its direct effect on the circular and longitudinal muscles of the intestinal wall. A decrease in
the motility of gut muscles increases the amount of time substances stay in the intestine [26]. This
allows for more water to be absorbed out of the water. We therefore presume that the reduction in
the intestinal propulsive movement in charcoal meal model may be due presence of similar
antispasmodic properties of plant extracts. It may be mentioned here that C. citratus possesses
essential oil, citral as one of its major active chemical components [16]. It is advocated that the
plants that have essential oils, are generally used traditionally for gastrointestinal disorders. In
several studies on relaxant effects of essential oils, including citral it has been reported that the
inhibition of contractile over-activity or reduction of inflammatory response of the ileum is their
basis for the treatment of gastro-intestinal disorders such as, diarrhoea [27, 28, 29, 30].
In the acute toxicity study, the plant extract up to a dose of 3200 mg/kg did not cause any mortality
or any changes in body temperature and food and water intake in the animals. The preliminary
observations indicate that the plant extract is non-toxic in nature. In conclusion, this study provides
support to the folk medicinal use of stalk decoction of C. citratus in the treatment of diarrhoea.
Acknowledgements
The award of a Senior Research Fellowship by the Council of Scientific & Industrial Research,
New Delhi to Vareishang Tangpu is gratefully acknowledged. Partial financial grants under the
DRS-III Programme of UGC in the Department of Zoology, NEHU is also acknowledged.
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