Journal of Herbal Medicine and Toxicology 3 (1) 31-36 (2009)

ISSN : 0973-4643 Original Article

EFFECT OF BACOPAMINE, A POLYHERBAL FORMULATION ON

LEARNING AND MEMORY

S. Desai1, D. Korgaonkar1, P. Gadre2, A. Ghogare2

1 Department of Pharmacology, K. M. Kundnani College of Pharmacy, Jote Joy Building, Rambhau
Salgaonkar Marg, Cuffe Parade, Colaba, Mumbai- 400 005, India
2 SciTech Healthcare Pvt. Ltd. Patel Terrace, Jijamata Road, Pump House, Andheri (East), Mumbai, India
dipti_083@yahoo.com, +91 22 2216 4368
Received : Dec 30, 2008, Revised : 15th Jan 2008, Accepted : 20th Jan 2009
Abstract : In the present study, a polyherbal formulation was investigated for its
effect on memory and learning. In Elevated Plus maze and Shock-avoidance test
Bacopamine (100 and 200 mg/kg, p.o.) was administered for eight consecutive days
to young adult mice of either sex and rats respectively. Significant improvement in
learning and memory was observed with animals treated with Bacopamine as
compared to control. We also investigated the effect of Bacopamine on 5 days
performance of mice in Morris Water maze. Oral daily treatment with Bacopamine
(100 and 200 mg/kg, p.o.) significantly improved memory and learning. Piracetam
(100 mg/kg, p.o.) was used as a positive standard. These results are suggestive of
Bacopamine providing excellent cognition enhancement.
Keywords: Cognition, nootropic, Elevated Plus maze, Morris Water maze, shock-
avoidance test

INTRODUCTION
Loss of memory and disturbed cognitive functions are
major concerns in people afflicted with neurological
diseases worldwide. Memory is the natural
counterpart of learning; it is a necessary condition
for the behavioral change to be permanent. [1]. Poor
memory, lower retention and slow recall are common
problems in today’s stressful and competitive world.
Age, stress, emotions are conditions that leads to
memory loss, amnesia, anxiety, dementia or
Alzheimer’s disease. [2] Although various synthetic
drugs for the treatment are available, side effects
associated with them make their use limited. In the
recent years, there has been a rise in the interest of
scientific community and pharmaceutical laboratories
to explore the therapeutic benefits of herbs to improve
memory. Several plants have been reported to
possess nootropic activity.Memory is an organism’s
ability to store, retain, and subsequently retrieve
information. Learning is defined as a relatively
permanent. The use of plants for healing purposes
predates human history and forms the origin of much
modern medicine. [3] Using recent scientific
developments, the medicinal properties of plants have
been investigated throughout the world, due to their
potent pharmacological activities, low toxicity and
economic feasibility. [4] Ayurveda is most ancient and
yet widely practised medicinal system in India. Several
herbal remedies are used to improve memory and
learning. Many herbal drugs used in Ayurvedic
medicinal practice are generally in the form of multi-
herbal formulae. The underlying expectation is that
the herbs in combination will synergize each others
therapeutic effect and diminish adverse effects. [5]
Cognitive function depends upon exteroceptive and
interoceptive feedback mechanisms. Exteroceptive
system is concerned with sensory stimuli such as

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visual, shock; whereas an interoceptive system deals
with physiological condition of the body. [6] In the
present study, the memory enhancing effect of
Bacopamine, a polyherbal formulation was evaluated
using exteroceptive behavior models. Spatial learning
was tested using Elevated Plus Maze and Morris
Water Maze models. Pole climbing apparatus was
used for shock-avoidance test.
MATERIALS AND METHODS
Drugs
The constituents of BacopamineR (BC) consist of
Bacopa monnieri, Cuminum cyminum, Curcuma
longa, Camellia sinensis, Menta piperita and Piper
nigrum. BC was obtained from SciTech Healthcare
Pvt. Ltd., Mumbai, and suspended in the distilled water
for oral administration. Piracetam (Pirament®, IPCA
Laboratories, India) was suspended in distilled water
and administered orally. Volume of oral administration
was 1 ml/100g of body weight.
Animals
Swiss albino mice of either sex in weight range of
20-25 g supplied by Haffkine Laboratory, Mumbai,
were used for Elevated Plus Maze and Morris Water
Maze models. Albino wistar rats of either sex (150-
200 g of body weight) were obtained from Bharat
Serum and Vaccines, Thane and used for shock
avoidance test.
The animals were grouped and housed in polyacrylic
cages, with not more than six animals per cage. They
were housed under standard conditions of
temperature and light and allowed free access to
standard dry pellet diet ( Amrut Laboratory animal
feed diet, Maharashtra) and water ad libitum. The
animals were acclimatized to laboratory conditions
for 5 days before commencement of experiments.
All experimental protocols were extensively reviewed
and approved by the Institutional Animal Ethics
Committee (IAEC) and care of animals was taken
as per guidelines of CPCSEA, Department of Animal
Welfare, Government of India.
Elevated Plus Maze
The elevated plus maze for mice consisted of two
open arms (25cm x 5 cm) and two covered arms
32
(25cm x 5 cm x 12 cm) extended from central
platform (5 cm x 5 cm), and the maze was elevated
to height of 25 cm from the floor. On the first day,
each mouse was placed at the end of an open arm,
facing away from the central platform. Transfer
latency (TL) was defined as the time (in seconds)
taken by the animal to move from the open arm into
one of the covered arm with all its four legs. TL was
recorded on the first day (training session) for each
animal. If mouse did not enter into one of the covered
arms within 90 seconds, it was gently pushed into
one of the two covered arms and the TL was assigned
as 90 seconds. The mouse was allowed to explore
the maze for another 2 min and then returned to its
home cage. Retention of this learned-task (memory)
was examined 24 hr after the first day trial. Significant
reduction in TL value on the second day indicated
improvement in memory. [2, 7-12]
Experimental Design:
Mice of either sex were divided randomly into four
groups of six animals each.
Group I: It represented the control group (n=6).
Distilled water (DW), was administered orally for 8
days. TL was noted after 90 min of administration on
eighth day and again after 24 hrs, i.e. on the ninth
day.
Group II: BC (100 mg/kg, p.o.) was administered
once daily for 8 days. On the eighth day, TL was
noted 90 min after the last dose and again after 24
hrs, i.e. on the ninth day.
Group III: BC (200 mg/kg, p.o.) was administered
once daily for 8 days. On the eighth day, TL was
noted 90 min after the last dose and again after 24
hrs, i.e. on the ninth day.
Group IV: Piracetam (100 mg/kg, p.o.) was
administered once daily for 8 days. On eighth day,
TL was noted after 90 min of dose administration
and again after 24 hrs, i.e. on the ninth day.
Morris Water Maze
This task was adapted for mice from the paradigm
originally described by Morris. [13] The water maze
was a circular pool (45 cm in diameter, 26 cm in
height), filled with water (26 ± 10 C) and made opaque
with white color, to the depth of 20 cm.

The pool was divided into four quadrants. An escape
platform was placed in the middle of the pool, 1.0 cm
below the water surface, equidistant from the sidewall.
Four different starting points for mice were placed
around the perimeter of the pool.
On each of the five training days, all four start points
were used once in a pseudorandom sequence. The
water maze was always located on a large room with
a number of extra-maze visual clues. The investigator
always sat at the same position.
The trial began by placing the animal in the water
facing the wall of the pool at one of the starting points.
If the animals failed to escape on the platform within
120 s, it was gently placed there by the researcher
and allowed to stay for 30 s. [14] The inter-trial interval
was 5-10 min. Four escape trials were given to all
mice per day for 5 consecutive days. The escape
latency (s) to reach the platform was recorded. [15,
16]
Experimental Design:
Mice of either sex were divided randomly into four
groups of six animals each.
Group I (Control group) was administered distilled
water, Group II& III were administered BC (100 and
200 mg/kg, p.o. respectively), Group IV was
administered Piracetam (100 mg/kg). Treatments for
all groups were once daily for 5 days. Latency time
(LT) was noted in all groups after 45 min of drug
treatment.
Shock avoidance Test
The apparatus consisted of a rectangular box (43 X
43 X 35 cm) with a pole (25 cm long and 3-cm
diameter) suspending vertically from the lid. The floor
of the chamber consisted of metal bars (0.5-cm
diameter) through which electric shock stimulation
were delivered. The pole attached to the plexiglass
ceiling of the box was devoid of electrical shock.
The experiment was performed in different steps.
(1.) Familiarization: The animal was placed on the
platform, After 10 sec of exploration; it is returned
to the home cage.
(2.) Learning (acquisition): An unavoidable footshock
is applied once (Footshock: 50 Hz; 1.5 mA) and the
Desai S. et al.
animal was allowed to jump on the pole, and then
returned to the home cage.
(3.) Retention Test: 24 hrs after the learning trial the
animal was again placed on the platform and the time
taken by the animal to jump on the pole (latency) was
measured.
Experimental Design:
Rats were divided into 4 groups and each group
consisted of a minimum of 6 animals.
Group I: It represented the control group (n=6).
Distilled water (DW), was administered orally for 8
days. LT was noted after 45 min of administration on
eighth day and again after 24 hrs, i.e. on the ninth
day.
Group II: BC (100 mg/kg, p.o.) was administered
orally for 8 days. Last dose was given 45 min before
subjecting the animals to foot shock. LT was noted
on the eighth day and again after 24 hrs.
Group III: BC (200 mg/kg, p.o.) was administered
orally for 8 days. Last dose was given 45 min before
subjecting the animals to foot shock. LT was noted
on the eighth day and again after 24 hrs.
Group IV: Piracetam (100 mg/kg) was administered
orally for 8 days. LT was noted after 45 min of
administration on eighth day and again after 24 hrs,
i.e. on the ninth day.
STATISTICAL ANALYSIS
Results are expressed as Mean ± Standard Deviation
(S.D.). The data was analyzed by one-way analysis
of variance (ANOVA) followed by Dunnett’s t-test.
p<0.05 was considered significant. All statistic
analyses were made using the software InStat.
RESULTS
Elevated Plus Maze
TL measured on 2nd day in mice was shorter than
that on the first day in all groups, indicating ability of
the mice to recall the learned aspect in a lesser period
of time [Fig.1]. BC showed dose-dependent reduction
in TL on 2nd day in mice as compared to control group,
indicating significant improvement in memory.
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Table- 1: Effect of Bacopamine Capsule* on Latency Time (LT) of Mice Using Morris Water
maze

Latency Time (seconds)

Days Treatment groups

Control BC 100 mg/kg BC 200 mg/kg Piracetam 100mg/kg

1 120 120 100 ± 10.954 110 ± 12.649

2 102.167 ± 6.337 73.621 ± 22.6106* 23.04 ± 13.687** 25.42 ± 19.571**

3 100.333 ± 7.967 47.213± 9.053** 12.123 ± 2.735** 36.517 ± 32.588**

4 97.22 ± 17.976 69.243 ± 31.452 21.267± 12.631** 15.027 ±13.987**

5 102.667 ± 7.866 76.587 ± 46.121 6.583 ± 2.247** 16.18 ± 17.54**

Values are expressed as mean ± SD, for N=6. One-way ANOVA followed by Dunnett test. *p<0.05
compared with Control group, **p <0.01 compared with control group

Morris Water Maze DISCUSSION

Over the 5 days of water maze testing, mice in control
group exhibited longer swim latency time in
comparison with 2 groups receiving BC regimen
[Table 1]. On the first day, there was no significant
difference in LT. Swim latencies exhibited by all
groups of mice were gradually decreased from day 1
to day 5. Significant reduction was observed with 200
mg/kg BC treated mice. This decrease in swim
latencies over 5 test days indicated that all the mice
eventually learnt the task.
Shock avoidance Test
Significant decrease in latency time on 2 nd day
indicated improvement in memory. Significant
reduction in latency time were observed on 1st day
with BC at a dose of 200 mg/kg (p<0.01) and
Piracetam at dose of 100 mg/kg (p<0.01) as
compared to control group. On 2nd day, there was
significant improvement in latency as compared to
control group with BC treated groups, 100mg/kg
(p<0.05) and 200mg/kg (p<0.01) respectively and
Piracetam treated group (p<0.01) as shown in Fig. 2.
BC (100 and 200 mg/kg) treatment was shown to
augment both the acquisition as well as the retention
of memory of learned task as seen by reduction in
TL as compared to control on Day 1 and Day 2
respectively.
In traditional practices of medicine, various plants
have been used to treat many memory related
disorders. An ethnopharmacological approach has
provided potential new drugs from plant sources,
including those for memory disorders. [17] The herbal
medicines have become the domain of ‘old wine
tables’. Nevertheless, in spite of their efficacy, herbal
medicines continue to reflect poor quality control both
for materials as well as clinical efficacy. [18]
Numerous animal models have been developed to
better understand mechanisms involved in learning
and memory. Several of these assess the ability of
rodents to recognize and remember a given location
in space and to relate this information to a specific
goal. This is often referred to as spatial learning. [19]
Arousal associated with the occurrence of some event
preferentially modulates the memory storage of
information related to that event. Adrenal
catecholamines and peptide hormones released into
systemic circulation during aroused states such as
electrical shock modulate information storage. [20]
Therefore, Shock-avoidance test can provide a useful
model to assess cognition. In the present study of
Bacopamine learning and memory was assessed
using Elevated Plus maze, Morris Water maze and
Shock avoidance Test. Treatment of BC improved
memory and learning in all three models. Difference
in TL in the control and test groups was observed in

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demonstrated that drugs that block muscarinic

Elevated Plus maze on 1st day, which might be related
acetylcholine receptors cause impairment in memory.
to the anti-anxiety effect of the drug. The
[21] Antioxidants help to neutralize free radicals.
neuromodulator acetylcholine plays an important role
These free radicals scavenge their missing electrons
in memory. Numerous studies in human subjects have
from other molecules and in doing so may cause

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Journal of Herbal Medicine & Toxicology
damage to important cell components. This oxidative
stress may be responsible for age-related neuronal
damage and neurodegerative disease. If antioxidants
counter the damaging free radicals, then it possibly
improves memory. [22] Mechanism of action of
Bacopamine is not known. But looking at the contents
of this formulation, we can say that it might exert its
antiamnestic effects by combination of its antioxidant
and anticholinesterase activities. Contents of the
Bacopamine Capsule, Bacopa monnieri [23],
Camellia sinensis [24,25], Curcuma longa [26],
Mentha piperita [27] and Piper nigrum [28] have
well established anticholinesterase activities. The
antioxidant activity of Bacopa monnieri [23],
Camellia sinensis [29], Cuminum cyminum [30],
Piper nigrum [31] and Curcuma longa [32] are
reported. They have an effective role in improvement
of memory.
The present study is thus indicative of Bacopamine
Capsule’s potent cognition enhancing properties. The
results substantiate the claim for the usage of
Bacopamine for short-term memory loss. Further
investigations are required to characterize the active
constituents responsible for improvement in memory
and learning.
REFERENCES
[1] Vervliet B.: Acta Psychologica, 127(3): 601-613 (2008).
[2] Parle M., Vasudevan M.: Journal of Health Science,
53(1): 43-52 (2007).
[3] Vickers A., Zollman C.: BMJ, 319: 1050-1053 (1999).
[4] Mukherjee B., Auddy B., Ferreira M., Blasina F., Lafon
L., Arredondo F. et al.: J. Ethnopharmacol., 84:131-
138 (2003).
[5] Andrade C., Chandra J. S.: Ind. J. Psychiatry, 48:232-
237 (2006).
[6] Hurliman E., Nagode J. C., Pardo J. V.: The Journal of
Neuroscience, 25(18):4641– 4648 (2005).
[7] Dhingra D., Parle M., Kulkarni S. K.: Ind. J. Pharm.
Sci.: 216-221(2006).
[8] Parle M., Dhingra D.: J. Pharmacol. Sci., 93: 129-135
(2003).
[9] Vasudevan M., Parle M.: Biol. Pharm. Bull., 29(6):
1154-1161 (2006).
[10] Raghavedra V., Kulkarni S. K.: Free Radical Biology
& Medicine, 30(6): 595-602 (2001).
36
[11] Kulkarni S. K., Verma A.: Indian Journal of Physiology
and Pharmacology, 36(1): 29-37 (1992).
[12] Kulkarni S. K., Verma A.: Indian Drugs, 30(3): 97-105
(1993).
[13] Morris R. Journal of Neurosciences, 11: 47-60 (1984).
[14] Fadda P., Robinson L., Fratta W., Pertwee R.G., Riedel
G.: Behav. Brain Res., 168:307-311 (2006).
[15] Enomoto T., Ishibashi T., Tokuda K., Ishiyama T.,
Toma S., Ito A.: Behav. Brain Res., 168(2): 197-207
(2008).
[16] Rubio J., Dang H., Gong M., Liu X., Chen S. L.,
Gonzales G. F.: Food Chem. Toxicol., 45:1882-1890
(2007).
[17] Howes M. R., Perry N. S. L., Houghton P. J.: Phytother.
Res.,17:1-18 (2003).
[18] Patwardhan B., Vaidya A. D. B., Chorghade M.:
Current Science, 86(6):789-799 (2004).
[19] Fox G. B., Komater V. A., Buckley M. J., Browman K.
E., Pan J. B., Hancock A. A., et al. : Behav. Brain Res.,
159: 295-300 (2005).
[20] Clark K. B., Smith D. C., Hassert D. L., Browning R.
A., Naritoku D. K., Jesen R. A. : Neurobiology of
Learning and Memory, 70:364-373 (1998).
[21] Sherman S. J., Arti A., Hasselmo M. E., Stern C. E.,
Howard M.W.: Behavioral Neuroscience , 117(3):526-
539 (2003).
[22] McDaniel M. A., Maier S. F., Einstein G. O.: Nutrition,
19:957-975 (2003).
[23] Russo A., Borrelli F.: Phytomedicine, 12: 305-317
(2005).
[24] Kakuda T.: Biol. Pharm. Bull., 25(12): 1513 (2002).
[25] Egashira N., Ishigami N., Pu F., Mishima K., Iwasaki
K., Orito K. et al.: Phytother. Res., 22(1): 65-68 (2008).
[26] Mishra S., Planivelu K.: Annals of Indian Academy
of Neurology, 11(1): 13-19 (2008-).
[27] Moss M., Hewitt S., Moss L., Wesnes K.: Int. J.
Neurosci., 1118(1): 59-77 (2008).
[28] Joshi H., Parle M.: J. Tradit. Med., 22(2-3): 39-43
(2005).
[29] Higachi-Okai K., Yamazaki M., Ngamori H., Okai Y.: J
UOEH, 23(4): 335-344 (2001).
[30] Dhandapani D., Subramanian R., Nmasivayam N.:
Journal of herbs, spices and medicinal plants, 11(3):
127-139 (2005).
[31] Vijayakumar R. S., Surya D., Nalini N.: Redox Resp.,
9(2): 105-110 (2004).
[32] Maheshwari R. K, Singh A. K., Gaddipati J., Srimal R.
C.: Life sciences, 78: 2081-2087 (2006).