Journal of Medicinal Chemistry and Drug Discovery

Available online at http://www.jmcdd.com

October - November, 2014, Vol. 4, pp 14-25
ISSN: 2347-9027

Research Article
ANTI-DIARRHOEAL AND LEARNING AND MEMORY ENHANCING ACTIVITY OF
AERIAL PARTS OF BASELLA ALBA

Komati Pavani
Department of Pharmacology, St.Marys College of Pharmacy, Secunderabad, Telangana
Email id: pavanicoti@gmail.com

ABSTRACT
The plant Basella Alba (Basellaceae) is a small ever green and possesses a number of medicinal properties. The purpose of the present study was
to evaluate the anti diarrhoeal activity of the ethanolic and aqueous extracts of the aerial parts of Basella Alba, by using castor oil induced
diarrhea model. The ethanolic and aqueous extracts of aerial parts of this plant at graded doses was investigated for anti diarrhoeal activity in
terms of reduction in the rate of defaecation and consistency of faeces in Castor oil induced diarrhea. Effect was further evaluated on
gastrointestinal motility test with charcoal meal and Learning and Memory enhancing activity also evaluated by using piracetam drug as a
standard. The aerial parts extract showed significant inhibitory activity against castor oil induced diarrhea and also showed nootropic activity.
There was significant reduction in gastro intestinal motility by the charcoal meal test in rats. The results obtained by this study substantiate the
anti diarrhoeal and nootropic effects of the ethanolic or aqueous extract.

KEY WORDS: Diarrhoeal activity, AEBA, EEBA, Castor oil, gastrointestinal motility.

INTRODUCTION
Diarrhoea is the production of soft or watery stools by increased frequency of bowel movements. It may be defined
as the passage of more than 300ml of liquid faeces in 24 hours. This results in fluid and electrolyte loss that may
lead ultimately to death, particularly in young children. Pain, urgency, perianal discomfort and incontinence often
accompany it1. Diarrhoea is caused by either various infectious agents or chemical agents.Nootropics(memory
enhancers) are drugs or neutraceuticals, or functional foods that are purported to improve mental functions such as
cognition, memory, motivation, attention, intelligence and concentration2-4.

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Basella Alba commonly known as Indian spinach, belonging to the family Basellaceae. The aerial part(leaves, stem)
of the plant serve as edible plant(vegetable) in many parts of the world5. The leaf juice is a demulcent, used in cases
of dysentery. Stem and leaves are used as mild laxative, diuretic and antipyretic. Also used in constipation, poultice
for sores, urticaria and gonorrhea. It is also used in poultice local swellings, intestinal complaints etc6. Present study
was conducted to investigate the antidiarrhoeal effect of Basella alba.

MATERIALS AND METHODS

Collection and identification of Plant material
Collection and drying of plant material: The aerial parts of Basella alba were collected from village areas of Siddipet
and authentified by botanist Dr. Rasingham BSI, Hyderabad and the voucher samples are kept in the BSI herbarium
for reference (BSI/DRC/12-13/Tech. /436).
Preparation of different extracts (petroleum ether, chloroform, ethanolic and aqueous extracts)7:
The stems of Basella alba were dried in shade at room temperature then made into a fine powder . Then the powder
was extracted with petroleum ether, chloroform, ethanol and water successively by soxhlet apparatus, maintained at
50oC to get extracts. These extracts were stored in airtight containers in a refrigerator below 10oC. The extracts
were examined for their colour and consistency. Their percentage yield was calculated with reference to air-dried
powder sample used for the extraction.
Animal selection:
For Antidiarrhoeal activity Wistar albino rats of either sex weighing between 150-200g and adult Swiss albino mice
(20-25g) of either sex, obtained from Srivenkateshwara enterprises, Bangalore were selected. The animals were
acclimatized to standard laboratory conditions (temperature 252oC) and maintained on 12 hours light; 12 hours
dark cycle. They were fed with ad libitum. Before performing the experiment the ethical clearance was obtained
from institutional animal ethics committee (IEAC). IAEC No.769/2010/CPCSEA.
Acute toxicity study:
The acute toxicity of Basella alba extract was determined by using albino mice (20-25g) those maintained under
standard husbandry conditions. The animals were fasted 12h prior to the experiment and acute oral toxicity as per
OECD guideline no. 420 was determined.

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Animals were administered with single dose of 2000 mg/kg extract and observed individually with special attention
given during the first 3 hours and for a total of 14 days. During this period the mortality and/or the moribund status
of the animals were noted8.
Anti-diarrhoeal activity:
Castor oil-induced diarrhoea9:
In the present study albino mice of either sex weighing 20-25 gm were used. They are divided in to six groups of 6
animals each. The various groups were treated as follows:
Group -1: Control (Castor oil 0.5 ml)
Group II: Standard (Loperamide + castor oil 0.5 ml p.o.)
Group III: Ethanolic extract (200 mg/kg p.o. + castor oil 0.5 ml p.o.)
Group IV: Ethanolic extract (400 mg/kg p.o. + castor oil 0.5 ml p.o.)
Group V: Aqueous extract (200 mg/kg p.o. + castor oil 0.5 ml p.o.)
Group VI: Aqueous extract (400 mg/kg p.o. + castor oil 0.5 ml p.o.)
Subsequently, the vehicle, loperamide and extracts were administered, orally to groups of 6 mice 60 min before the
administration of the castor oil. The following parameter were determined, the time elapsed between the
administration of the cathartic agents and the excretion of the first diarrhoeic faeces (wet faeces that leaves a hallo
on the filter paper); the total number faeces as well as the number of diarrhoeic faeces excreted by the animals in 4
hrs; and the total weight of diarrhoeal stool in the period of time.
Gastrointestinal motility test10:
In the present study albino mice of either sex weighing 20-25 gm were used. They are divided in to 6 groups of 6
animals each. The various groups were treated as follows:
Group I: Control (Charcoal meal 0.5 ml p.o.)
Group II: Standard (Atropine sulphate i.p. +

Charcoal meal 0.5 ml p.o.)

Group III: Ethanolic extract (200 mg/kg p.o. + Charcoal meal 0.5 ml p o.)
Group IV: Ethanolic extract (400 mg/kg p.o. + Charcoal meal 0.5 ml p.o.)

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Group V: Aqueous extract (200 mg/kg p.o. + Charcoal meal 0.5 ml p.o.)
Group VI: Aqueous extract (400 mg/kg p.o. + Charcoal meal 0.5 ml p.o.)
Subsequently, after 30 min each individual mouse was administered with 0.5 ml of a 5% charcoal suspension in a
10% aqueous suspension of tragacanth mucilagel was administered p.o. to each animal.
After 30 min of this treat, each rat was sacrificed and intestinal distance moved by the charcoal meal from pylorus to
cecum. The percentages of distance travelled by the charcoal meal in ratio to the intestinal length and percentage of
inhibition was calculated by using following formula.

Distance travelled by the charcoal meal % travelled =

x 100

Total length of small intestine

(Total length of the small intestine Distance travelled by the charcoal meal) % of inhibition = x 100
Total length of small intestine
Learning and Memory enhancing activity11-21:
Experimental Procedure: Laboratory model for testing learning and memory:
Elevated plus maze test: The elevated plus maze was described as tool for testing memory by the investigator
working in the field of psychopharmacology. Elevated plus maze served as extroceptive behavioral model to
evaluate learning and memory in rats. The elevated plus maze consists of two open arms and two closed arms
(50cmx10cmx40cm) with an open roof arranged so that the two arms are opposite to each other. The maze was
elevated to a height of 50 cm.
Model A: Effect of extracts on Transfer ratio in Diazepam induced amnesia:
Mice were divided into 7 groups consisting of 6 animals per group. Group-1 treated with Normal control (Distilled
water 10ml/kg, p.o) only once daily for 14 days, Group-2 treated with Diazepam (5mg/kg, i.p) alone on first day
only and after 45 mins, TL was recorded on EPM and retention (memory) of learned task was observed 24 hrs later.
The TL was calculated as described above. Group-3 treated with standard drug piracetam (200 mg/kg, p.o). Groups

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3, 4 and 5, 6 were treated orally with different doses EEBA (200 & 400 mg/kg) and AEBA (200 & 400 mg/kg)
respectively only once daily for 14 days.
On the 14th day, 90 minutes of treatment of last dose, TL is recorded. Twenty four hours later i.e., on 15th day
transfer latency was again recorded.
Statistical analysis:
All results will be expressed as mean SEM from 6 animals. Statistical difference in mean will be analyzed using
one-way ANOVA (analysis of variance) followed by Post hoc test (Dennettst test). P< 0.05*, 0.01** and
0.001*** will be considered as statistically significant.
RESULTS & DISCUSSION
The % yields of Aqueous and Ethanolic etracts are 10.0 and 8.0 respectively. The preliminary phytochemical
analysis of petroleum ether, chloroform, ethanolic, and aqueous extracts of aerial parts of Basella alba revealed the
presence of various phytoconstituents (table-1)
Acute toxicity test
The mice treated with EEBA and AEBA at a dose of 2000 mg/kg, p.o. exhibited normal behaviour, without any
signs of passivity, stereotypy and vocalization. Their motor activity and secretory signs were also normal and no
sign of depression. EEBA and AEBA even up to the dose level of 2000 mg/kg body weight did not produce any
behavioural symptoms or mortality. So 1/10th and 1/5th doses of (maximum dose tested for each extract) were
selected as medium and high doses and were used in the present study to explore anti diarrhoeal and learning and
memory enhancing activities.
Anti-diarrhoeal activity:
Castor oil-induced diarrhoea:
Effect of ethanolic and aqueous extracts of aerial parts of Basella alba on castor oil induced diarrhoea:
All the parameters of castor oil induced diarrhoea (total no. of faeces, on set time of diarrhea, no. of wet feces and
total weight of wet feces (fig.1-3)) were reduced by treatment of ethanolic extracts and aqueous extracts of aerial
parts of Basella alba where as the time required for diarrhoeal episode have been significantly reduced by the above
mention extracts as well as the standard drugs (loperamide). Between the two extracts ethanolic extract was found to
be more potent (table-2).

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Table1: Phytochemical evaluation of different extracts of Basella alba
S.No.
1
2
3
4
5
6
7

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Tests
Alkaloids
Carbohydrates
Flavonoids
Saponins
Sterols
Tannins
Glycosides

Petroleum ether
-ve
-ve
-ve
-ve
+ve
-ve
-ve

Chloroform
-ve
-ve
-ve
-ve
+ve
-ve
-ve

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Ethanol
+ve
+ve
+ve
+ve
+ve
+ve
+ve

Water
+ve
+ve
+ve
+ve
+ve
+ve
+ve

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Table No.2: Effect of EEBA & AEBA on castor oil induced diarrhoea

Treatment Dose (mg/kg)

Control
Standard

0nset time of Total

diarrhoea
number of
(min)
faeces
103.51.8
9.161.13
203.50.4**
4.230.73**

Number of
wet faeces
6.330.80
3.450.61**

Total weight
of wet faeces
(mg)
35353.89
169.166.83**

140.51.8*
195.51.9**

5.500.56*
5.021.07**

4.150.49*
3.631.01**

198.535.44*
178.2314.04**

EEBA
EEBA

Distilled water
Lopera-mide
(3mg/kg)
200 mg/kg
400 mg/kg

AEBA

200 mg/kg

132.50.8

6.030.77*

5.031.05

223.1636.91

AEBA

400 mg/kg

135.56.6*

5.830.54*

4.531.05*

221.6614.87*

Values are expressed as mean S.E.M., n=6, significant at *P < 0.05 and **P < 0.01, When compared to
control group. Standard drug: Loperamide (3mg/kg).
Fig.No.1: 0nset time of diarrhoea in castor oil induced diarrhea

Time in min

250
200
150
100

0nset time of
diarrhoea (min)

50
0

Groups

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Fig.No.2: Total number of faeces in castor oil induced diarrhea

Number of faeces

12
10
8
6

Total
faeces

2
0

wet faeces

Groups

Fig.No.3: Total weight of wet faeces in castor oil induced diarrhoea

Total weight of
wet faeces

600
400
200
0

Total weight of wet

faeces

Groups
Gastrointestinal motility:
Effect of ethanolic and aqueous extracts of aerial parts of Basella alba on Gastrointestinal motility:
Pre-treatment with ethanolic and aqueous have

reduced the GIT motility in a dose dependent manner by the reduction in the movement charcoal meal.
Between the two extracts ethanolic extract was found (table-3) to be most potent.

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Table No.3: Effect of EEBA & AEBA on Gastrointestinal motility
Treatment

Dose(mg/kg)

Distance traveled by charcoal meal

with SEM (%)

Control

Distilled water

91.254.76

Standard

Atropine sulphate
(0.1 mg/kg)

56.102.30**

EEBA

200 mg/kg

64.963.56*

EEBA

400 mg/kg

61.682.38**

AEBA

200 mg/kg

77.054.58

AEBA

400 mg/kg

72.834.0*

S.NO
1

Values are expressed as mean S.E.M., n=6, significant at *P < 0.05 and **P < 0.01, When compared to
control group. Standard Drug: Atropine sulphate (i.p)

Distance traveled by
charcoal meal with SEM
(%)

Fig.No.4: Distance travelled by charcoal meal in Gastrointestinal motility

120
100
80
60
40
20
0

Distance travelled by
charcoal meal

Groups

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Learning and memory enhancing property:
The ethanolic and aqueous extracts of aerial parts of Basella alba were studied for the learning and
memory enhancing properties. The results explain the transfer latencies (TL) of the various drug treated
groups i.e., control, toxic control, standard, ethanolic and aqueous extracts of the extracts of aerial parts of
Basella alba(table-4).
Table No 4: Nootropic effect of aerial parts of Basella alba in mice with EPM model.
Treatment

Dose (per Kg)

Transfer Latency
14 day
15 day

I.

Normal control

10 (ml,p.o)

35.693.25

33.51.82

II.

5 mg/kg

51.782.60

583.266

III.

Toxic control
Piracetam
(200mg/kg)

200 (mg, p.o)

20.602.72

10.452.02**

IV.
V.
VI.
VII.

EEBA
EEBA
AEBA
AEBA

200 (mg, p.o)

400 (mg, p.o)
200 (mg, p.o)
400 (mg, p.o)

22.332.02
41.431.78
25.321.97
43.333.65

13.831.887**
26.531.86*
15.762.39**
33.822.24*

Group
No.

Values are expressed as mean S.E.M., n=6, significant at *P < 0.05 and **P <0.01, when compared to
Toxic control.Values are expressed as mean S.E.M., n=6, significant at *P < 0.05 and **P <0.01, when
compared to Toxic control.

CONCLUSION
In the present study the aerial parts of Basella alba was subjected to phytochemical and pharmacological evaluation.
Both the extracts were subjected to screening for anti - diarrhoeal activity by experimentally induced diarrhoeal
models such as Castor oil induced diarrhoea, Gastro intestinal motility test.In castor oil induced model,
administration of castor oil increase the number of faeces, no. of wet faeces and total weight of wet faeces, it
reversed by pre treatment with ethanolic and aqueous extract significantly. There were reports that tannins
precipitate proteins responsible for anti- diarrhoeal property. The exact mechanism of reduction in GIT motility due
to pre - treatment with both the extracts was not known. However it seems this direct action on GIT motility is
having a most important role in anti - diarrhoeal effects.In case of memory enhancing activity transfer latency (TL)
of first (7) day reflected learning behavior of animals whereas, TL of Second (8) day reflected retention of
information or memory. Basella alba (200 & 400 mg/kg) of both extracts and piracetam (200 mg/kg) administration
for 7 days orally significantly decreased TL on first day as well as second days, indicating significant improvement

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of learning and memory.Finally concluded that, Both the ethanolic and aqueous extracts of the plant possess antidiarrhoeal property as indicated by the reduction in the total weight of wet feces, in the experimental models.
Between two extracts ethanolic extract produced most potent anti-diarrhoeal activity. Saponins contribute to the
learning and memory enhancing property. The presence of saponins in ethanolic and aqueous extracts, after
administration of these extracts there was decreased TL on first day as well as second days, indicating significant
improvement of learning and memory. The further research on the plant will be definitely a contributory for
pharmacological establishment.

REFERENCES
1. Regi RK and Kuttan R; Inhibition of experimental gastric lesion and inflammation by Phyllanthus amarus extract.
Journel of Ethnopharmacol; 2003; 87: 193-97.
2. Goodman and Gilman; The pharmacological basis of therapeutics. 9th edition. New York; Mc Graw-Hill 1996:
376.
3. Chaudhuri SK; Quintessence of medical pharmacology. Calcutta: New central book agency Ltd. 1997: 230.
4. Jagdeep SD, Prasad DN, Avinash C. Tripathi and Gupta R; Role of Traditional Medicine in Neuro
psychopharmacology; Asian Journal of Pharmaceutical and Clinical Research; 2009; 2(2):72-79.
5.David Wilson et al.; Invitro Screening For Antiinflammator Activity Of Basella Alba And Alternanthera Sessilis
An Important Edible Green Leaves; Intl. J. Comphren. Res. Bio. Sci; 2014;1(1): 25-28.
6.Roshan A, Naveen KHN and Shruthi SD; A Review on Medicinal Importance of Basella alba L. International
Journal of Pharmaceutical Sciences and Drug Research; 2012; 4(2): 110-14.
7. Evans WC and Trease; Text Book of Pharmacognosy. ELBS, London, 1999; 14th Edition: 227-261.
8. OECD-Guidelines for Testing of Chemicals, Revised Draft 420, Documents on Acute Oral Toxicity and Acute
Toxicity Class Method; 2001.
9.Galvez J, Crespo ME, Jimenez J, Suarez A and Zarzeulo A; Antidiarrhoeic activity of quercitrin in mice and rats.
Journal of Ethanopharmacol; 1993; 45:157-59.
10. Mujumdar AM, Misar and Upadhye AS; Antidiarrhoeal activity of ethanol extract of the bark of Dalbergia
lanceolaria; Journal of Ethnopharmacol; 2005; 102: 213-16.

ISSN:

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Page 24

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11. Chattipakorn S, Pongpanparadorn A and Pratchayasakul W; Tabernaemontana divaricata extract inhibits
neuronal acetyl cholinesterase activity in rats, J. Ethnopharmacol. 2007; 110(1):61-68.
12. Gorman M, Neuss W, Cone NJ and Deyrup JA; Alkaloids from Apocynaceae.1960.
13. Pratchayasakul W, Pongchaidecha A, et al; Ethnobotany & ethnopharmacology of Tabernaemontana divaricata,
Indian journal of medical research; 2008; 127: 317-335.
14. Raj K, Shoeb A, Kapil RS and Popli SP; Alkaloids of Tabernaemontana divaricata. Phytochem; 1974; 13:162122.
15. Chattipakorn S, Pongpanparadorn A and Pratchayasakul W; Tabernaemontana divaricata Extract Inhibits
Neuronal Acetyl cholinesterase Activity in Rats. Jounel of Ethnopharmacol; 2007; 110: 61-8.
16. Hoernle AFR; The Bower manuscript (Archaeological Survey of India, New Imperial Series), Superintendent of
Government Printing, India, Calcutta, 1893-1912; 22:18-188.
17. Pawelka and Stoeckigt KH; Indole alkaloids from cell suspension cultures of Tabernaemontana divaricata and T.
iboga, Journel Plan of Cell Report; 1983; 22:105-7.
18. Preksha D et al; A traditional approach to herbal nootropic agents: An overview; International Journal of
Pharmaceutical Sciences and Drug Research; 2012; 3(3): 630 -636.
19. Vasudevan M and Parle M; Pharmacological actions of Thespesia populnea revelant to Alzheimers disease,
Phytomedicine; 2006; 13:677-687.
20. Jaiswal AK and Bhattacharya SK; Effects of Shilajit on memory, anxiety and brain monoamines in rats, Indian
journal of Pharmacology; 1992; 24: 12-17.
21.Walika N, Khongsombat, Onrawee, Pornnarin, Niwat and Kornkanok; The effects of Tabernaemontana
divaricata; root extract on amyloid -peptide 2535 peptides induced cognitive deficits in mice, Journal of
Ethnopharmacology; 2010; 130(1):122-12.

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