phytochemical evaluation and pharmacological … · phytochemical evaluation and pharmacological...
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Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
PHYTOCHEMICAL EVALUATION AND PHARMACOLOGICAL
SCREENING OF ETHANOLIC LEAF EXTRACTS OF EICHHORNIA
CRASSIPES AND NELUMBO NUCIFERA FOR
NEUROPHARMACOLOGICAL ACTIVITY IN PSYCHONEUROSIS
INDUCED MICE
Mehnoor Farheen*1, Syeda Arman R. O. Hussaini
2, Asfia kouser
3
1*
Department of Pharmacology, Assistant Professor Shadan womens College of Pharmacy,
JNTU, Hyderabad.
2,3M. Pharm [Pharmacology], Shadan Womens College of Pharmacy, JNTU, Hyderabad.
ABSTRACT
Behavioral neuropharmacology focuses on the study of how drugs
affect behavior in humans and animals. This study assessed the
sedative, CNS depressant, Analgesic, Anti-epileptic, Anti-anxiety,
Anti-psychosis, Anti depressant and Memory enhancing properties of
ethanolic extract of Nelumbo nucifera, Eichhornia crassipes and
combination of ethanolic extracts of Nelumbo nucifera and Eichhornia
crassipes in disease induced mice. Extraction of ethanolic extract from
leaves of Nelumbo nucifera and Eichhornia crassipes was performed
by maceration. 24 mice were divided into six groups. Group I
consisted of normal mice that were given only sterile saline solution
and served as control group. Group II consisted of mice induced with
depression, anxiety, psychosis, epilepsy and were treated with Normal
saline. Group III, IV and V consisted of normal mice induced with depression, anxiety,
psychosis, epilepsy. Oral administration of extract resulted in significant sedative, CNS
depressant, Analgesic, Anti-epileptic, Anti-anxiety, Anti-psychosis, Anti depressant and
Memory enhancing effects. Group VI consisted of normal mice induced with depression,
anxiety, psychosis, epilepsy and were treated with standard drug. The effects produced by the
extracts were closely similar to the standard drugs. In conclusion, the present study indicates
that the ethanolic leaf extracts of Nelumbo nucifera and Eichhornia crassipes and
WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
SJIF Impact Factor 5.210
Volume 4, Issue 12, 874-904 RReesseeaarrcchh Article ISSN 2278 – 4357
Article Received on
05 Oct 2015,
Revised on 29 Oct 2015,
Accepted on 22 Nov 2015
*Correspondence for
Author
Mehnoor Farheen
Department of
Pharmacology, Assistant
Professor Shadan womens
College of Pharmacy,
JNTU, Hyderabad.
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Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
combination of ethanolic extracts of Nelumbo nucifera and Eichhornia crassipes appears to
exhibit sedative, CNS depressant, Analgesic, Anti-epileptic, Anti-anxiety, Anti-psychosis,
Anti depressant and Memory enhancing activity in disease induced mice.
KEYWORDS: Anti-psychosis, CNS Depressant, Depression, Eichhornia crassipes, Memory
enhancing, Neuropharmacology.
1. INTRODUCTION
Neurological disorders cause many physical, mental, emotional and social stresses to
individuals suffering from them. Indivituals suffering from a neurological disorder or
disorders may experience many serious, long-term mental complications. These
complications give rise to social inadequacies which can gradually worsen with time. Mental
health is an important issue which is not given enough attention, thereby increasing the
number of mental health patients by the day. Data has shown a disturbing increase in
numbers of mental health issues in just 5 years (2005 to 2010).[1]
Mental disorders are the
core health challenge of the 21st century. Most Neurological disorders are preventable and
easily treatable with drugs. These drugs come with a number of complications associated
with them which can worsen the patients mental health and be very pricey, causing patients to
avoid over-all therapy. Patients search for natural substances which can give relief as well as
be cost-effective, this is where researches such as this one, come into play. Medicinal plants
have attracted global interest in recent years. In the past few decades, pioneer work in
identification, documentation and recognition of traditional medicine has been done in India.
Investigation of traditional medicine is very important for the welfare of rural and tribal
communities for the treatment of conventional illness. In this study, Nelumbo nucifera,
Eichhornia crassipes and combination drug consisting of both Nelumbo nucifera and
Eichhornia crassipes was evaluated for its use as a sedative, CNS depressant, Analgesic,
Anti-epileptic, Anti-anxiety, Anti-psychosis, Anti depressant and Memory enhancing
substance.
Review of Drug under study
Nelumbo nucifera
Nelumbo nucifera, also known as Indian lotus is one of two species of aquatic plant in the
family Nelumbonaceae. In Ayurveda Nelumbo nucifera is used as a diuretic and anthelmintic
and in the treatment of strangury, vomiting, leprosy, skin diseases and nervous
exhaustion.[2,3,4]
In popular medicine it is used in the treatment of tissue inflammation, cancer,
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skin diseases, leprosyand as a poison antidote.[5,6]
Rhizomes are prescribed asdemulcents for
haemorrhoids and are beneficial in dysentery,chronic dyspepsia, and have nutritive, diuretic
and cholagogueactivities.[7,8]
The leaves are usedfor the treatment of haematemesis, epistaxis,
haemoptysis,haematuria, metrorrhagia and hyperlipidaemia.[9]
The flowersare useful in the
treatment of diarrhoea, cholera, fever and gastriculcers[5]
The seeds and fruits are used as a
health food in Asia and to treat many ailments, including poor digestion, enteritis, chronic
diarrhea, insomnia, palpitations, spermatorrhoea, leucorrhoea, dermatopathy, halitosis,
menorrhagia, leprosy, tissue inflammation, cancer, fever and heart complaints, and as
anantiemetic, poisoning antidote, diuretic and refrigerant.[5,10,11,12]
Lotus seedpods are
sometimes used as a traditional medicine for haemostatic function.[13]
The seed powder
mixed with honey is useful in treating cough.[3]
Embryos of lotus seed are used in traditional
Chinese medicine to overcome nervous disorders, insomnia, high fevers (withrestlessness)
and cardiovascular diseases (e.g. hypertension, arrhythmia).[14]
Eichhornia crassipes
Eichhornia crassipes, commonly known as (common) water hyacinth, is an aquatic plant
native to the Amazon basin, and is often considered a highly problematic invasive species
outside its native range. This invasive nuisance is planta non grata in much of the world
where it often jams rivers and lakes with uncounted thousands of tons of floating plant
matter. A healthy acre of water hyacinths can weigh up to 200 tons. the water hyacinth in
most places is under "maintenance control": field crews constantly working to keep the plant
numbers at their lowest possible levels, in exchange for the rivers and lakes remaining usable.
Eichhornia crassipes is used for Bioenergy, phytoremediation, food and as animal feed and
fertilizer. Studies show, it can be used as Anti-algal [15]
, Anti-Microbial [15]
and
Antioxidant.[16]
This research has tested the efectivity of the two plants, Nelumbo nucifera
and Eichhornia crassipes, for their neuropharmacological property. The findings, were
impressive.
MATERIALS AND METHODS
Collection of drugs
Dried leaves of both plants Eichhornia crassipes (Pontederiaceae) and Nelumbo nucifera
(Nelumbonaceae) were collected. The plants were taxonomically identified and authenticated
by DR. K. MADHAVA CHETTY Assistant Professor of Botany, Department of
Pharmacognosy, Sri Venkateshwara University, Tirupathi.
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Preperation of plant extract
The leaves were cut into pieces and subjected to shade drying. On complete drying, the
pieces were powdered and stored in air tight containers at room temperatures. The powdered
was macerated with ethanol for 7 days and then filtered. The filtrate was evaporated to obtain
dried extract. The extract thus obtained was subjected for evaluation of neuropharmacological
and analgesic activity study. The plant extract was prepared by maceration process.
Maceration: In maceration(for fluid extact), whole or coarsely powdered plant-drug is kept in
contact with the solvent in a stoppered container for a defined period with frequent agitation
until soluble matter is dissolved. This method is best suitable for use in case of the
thermolabile drugs. Using this process, 500g/kg powder was added in ethanol in the ratio 1:2
and vigorous shaking was carried out for 7 days continuously and was kept at room
temperature. The filtrate thus obtained was ethanolic extract. The filtrate obtained was
dissolved in 0.9% normal saline and used as vehicle in the experiment.
Preliminary phytochemical screening
Standard screening test of the extract was carried out for various plant constituents. The crude
extract was screened for the presence or absence of secondary metabolites such as in
Alkaloids, flavonoids, sterols,Glycosides, terpenoids, anthroquinone, proteins, phenols and
anthocyanins using standard procedures.
Acute toxicity studies
Acute toxicity study was performed for the extract according to the acute toxic classic
method as per the method of Litchfield & Wilcoxon (1949). Acute toxicity study was carried
out on plant extracts using Male and Female Albino mice. The mice were fasted overnight
and the weight of each mouse was recorded just before use. The animals were divided into
five groups containing 6 animals each, the extract was administered orally in increasing dose
up to 2000 mg/kg b.w. After the treatment the animals were observed for mortality or toxicity
for 72 hours. No changes in skin and fur, eyes, autonomic (salivation, lacrimation,
defecation) and central nervous system (ptosis, drowsiness, gait, tremors and convulsions)
were observed. No deaths were observed.
Experimental animals
Albino mice (15-50 gms) of either sex housed in standard conditions of temperature (55 ±
5%) and light (12 hrs light/dark cycles) were used. They were fed with standard pellet diet
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and water and libitum. Animals were randomly selected for grouping. All experiments were
performed according to the norms of the ethical committee (CPCSEA).
SCREENING METHODS
Head dip test
A specially designed rectangular-shaped head dip box having 15 holes in each side was used
in this study. The number of head dips by mice through these holes in a specified time was
counted. Mice of groups II, III, IV received Test drug 1, test drug 2 and combination of test
drugs 1 and 2 respectively. The groups I and V received control and standard drug Diazepam
(2 mg/kg, i.p). The control and drug-treated animals were placed individually in the head dip
box and the observations were made for 30 min[17][18][19]
Locomotor Activity
It can be studied with the help of an Actophotometer. Each mouse was placed individually in
actophotometer & the basal activity score of the animals were recorded after 30, 60 & 120
mins of drug treatment. Groups III, IV, V received Test drug 1, test drug 2 and combination
of test drugs 1 and 2 respectively. The groups I, II and V received control, Normal saline and
standard drug Diazepam (3 mg/kg, i.p ). Activity on each mouse was retested for 10mins.
Difference in the activity was recorded considering before treatment values & after treatment
values. Finally, Percentage decrease in locomotor activity was Calculated[20]
Rota-rod test (Thread-mill device test)
The study was carried out according to the method described by Perez et al., (1998). Rota rod
treadmill device was used for this experiment. Mice trained to remain on slowly moving (16
rpm) rods of 5 cm diameter for 180 seconds or longer were selected. . Groups II, III, IV
received Test drug 1, test drug 2 and combination of test drugs 1 and 2 respectively. The
groups I and V received control and standard drug Diazepam (10 mg/kg, i.p ). One hour after
administration of test drugs, the mice were placed singly on the rod for 3 minutes, at 30
minutes intervals for 3 h. If an animal failed more than once to remain on the rod for 3
minutes, the test was considered positive, meaning that there is lack of motor coordinations.
Pentobarbitone induced sleep test in mice
The method of Rakotonirina et al. (2001), was adopted. Sleep potentiation time of the test
drugs was studied in group of mice that received Pentobarbitone at a dose of 30 mg kg body
weight one hour after i.p. administration of Teat drug 1(Group II), test drug 2(Group III),
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combination of both test drugs(Group IV), standard drugs Diazepam(Group V) and Normal
saline(Group I); with 4 mice in each group. The sleeping time was estimated as the time
between disappearance and recovery of the straightening reflex.
Hole cross test
The method was adopted as described by Takagi et al. (1971). A partition was fixed in the
middle of a cage having a size of 30 × 20 × 14 cm. A hole of 3 cm diameter was made at a
height of 7.5 cm in the centre of the cage. The number of passage of a mouse through the
hole from one chamber to the other was counted for a period of 3 min at 0, 30, 60, 90 and 120
min after oral administration of Normal saline(Group I), the test drugs(Group II, III), their
combination(Group IV) and standard drug( Diazepam 1mg/kg b.w)(Group V). (Takagi et al.,
1971).
Hot plate method
The method was adopted as described by Sachin Kumar Jain et al., 2014. The mice of either
sex were weighed and divided into five groups (n = 4 in each groups). Group I served as
control. Group V (Aspirin 10 mg / kg body weight) served as standard and group II, III and
IV were treated with test drug 1, test drug 2 and combination of test drugs 1 and 2
respectively. Reaction time of animals was noted down in hot plate at 0, 30, 60, 90 and 150
minutes after the treatment. The basal reaction time taken by observing hind paw licking or
jump response (whichever appear first) in animals while placed on hot plate,which was
maintained at constant temperature 55ºC ± 1ºC. A cut off period of 15 seconds was observed
to avoid damage to the paws. The percentage of analgesia was calculated.
Acetic acid-induced writhing in mice
The experiment was carried out according to the method of Witkin et al. Test drugs were
administered by the oral route one hour prior to the injection of acetic acid. Writhing was
induced in animals by injecting acetic acid 300 mg/kg (3% solution in sterile distilled water)
i.p. Each mouse was then kept indivitually and the total number of writhing episodes for a
period of 20 minutes after the injection of acetic acid was counted. The percent inhibition of
the writhing count of the treated group was calculated from the mean writhing count of the
control group.
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Tail flick test
Albino mice were fasted for 24 hrs with water maintained at room temperature. Mice were
treated by oral gavage with Normal saline(Group I), Test drug 1(Group II), test drug 2(Group
III), combination of test drugs 1 and 2(Group IV) and standard drug(Group V), Aspirin (100
mg/kg, i.p). Nearly 1 or 2 cm of the tail was immersed in warm water kept constant at 50 ºC.
The reaction time was the time taken by the mice to deflect their tails. The first reading was
discarded and the reaction time was taken as the mean of the next two readings. The latent
period of tail flick response was taken as the index of anti-nociception and was determined at
15, 30, 60 and 90 min after the administration of drugs. The maximum reaction time was
fixed at 15 sec (Sewel and Spencer., 1976).
Isoniazid Induced Convulsions
Isoniazid at the dose of 300 mg/kg was injected i.p. to induce clonic-tonic convulsions in
mice. Groups II, III, IV received Test drug 1, test drug 2 and combination of test drugs 1 and
2 respectively. The groups I and V received control, and standard drug Phenytoin (10 mg/kg,
i.p ). Each animal was placed in individual plastic cage for observation lasting forty five
minutes. The parameters noted were mean onset time of convulsions, duration of convulsions
and number of deaths (% mortality).
Elevated plus Maze test
Anxiety-related behavior was measured by the elevated plus-maze test. The elevated plus-
maze Consist of two open arms, 50 × 10 cm, and two closed arms, 50 × 10 × 40 cm. The
maze was elevated to a height of 15 cm above the floor. Groups II, III, IV, received Test drug
1, test drug 2 and combination of test drugs 1 and 2 respectively. The groups I and V received
control, Normal saline and standard drug Diazepam (35 mg/kg, i.p ). Each mouse was placed
on the central platform facing a closed arm. During 5 min test period, the following measures
were taken by an observer, number of open arms entries, time spent in open arms, number of
closed arm entries, time spent in closed arms. Entering into an arm was noted only when all
paws had crossed out central area. (Kurt, M.,2004).
Novelty Induced Hypophagia
Albino Mice were divided in to five groups (n=6). Three days was allowed for habituation by
exposing mice to sweetened milk (Carnation sweetened condensed milk, 1:3 in water) in the
home cage (~1-2 hr sessions). On the fourth day, latency to drink from the sweetened milk
bottles in their home cage (dim lighting) was noted. On the fifth day drug administration of
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Test drug 1, test drug 2 and combination of test drugs 1 and 2 and standard drug to mice of
groups II, III, IV and V respectively were done followed by measuring latencies to drink the
sweetened milk in a novel cage (bright lighting). Comparisons of drinking latencies between
home cage and novel cage were made.(Merali et al., 2004).
Evasion test
The method of Turner (1965) was used. The animals were introduced into a rectangularbox
with an inclined plane by which the mice can escape from the box, and the mice that escaped
within 5min from the rectangular box were selected for this test. 15 minutes after
administration of normal saline(Group I), diazepam(0.5mg/kg) (Group V) as standard and
Test drugs1(Group II), 2(Group III) and combination of both(Group IV), the animals were
placed in the box again and the number of mice remaining in the box after 15 min in each
group was noted.
Caffeine-induced stereotypy and locomotion
The method described by Randrup and Munkvad (1967) was used for the stereotype
behavioural studies. Adult mice were randomized into five groups of six mice each. Groups
III, IV, V received Test drug 1, test drug 2 and combination of test drugs 1 and 2 respectively.
The groups I, II and V received control, Normal saline and standard drug chlorpromazine (2
mg/kg, i.p ).. One hour after administration of saline and extract and thirty minutes after
chlorpromazine administration, all the mice were given 2 mg caffeine/kg. The signs of
stereotype behaviour that included circling, jumping, sniffing, and general locomotion were
recorded for a period of 2 h using a hand held tally counter (Irwin, 1968).
Stereotyped behavior was scored as follows.
-Complete absence of stereotyped behavior, presence of stereotyped movements of the
head and intermittent sniffing= 1,
-Sniffing and chewing= 2;
-Chewing and intense licking= 3.
Locomotion was assessed using the method of Sturgeon et al. (1979) simultaneously with the
stereotypy and scored as follows.
-Stationary= 0;
-Movements within a localized area, forelimbs only= 1;
-Intermittent movements within half of the area of the cage= 2;
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-Continuous movements within half of the area of the cage= 3;
-Intermittent movements within the whole area of the cage= 4;
-Continuous movements within the whole area of the cage= 5.
Catalepsy in mice
Five groups of 4 albino mice each weighing between 20-50g are used. They are treated with
the test drug or the standard given intraperitoneally i.e. Groups II, III and IV received Test
drug 1, test drug 2 and combination of test drugs 1 and 2 respectively. The groups I and V
received Normal saline and standard drug Resperidone (1 ml/kg, i.p ). After 30 min they are
placed individually into translucent plastic boxes with a dowel mounted horizontally 15cm
from the floor and 5cm from the end of the box. The floor of the box is covered with
approximate 8cm of the bedding material. The animals are allowed to adapt to the box for 5
min. Thereafter each animal is grasped gently around the shoulders and the forepaws are
placed carefully on the dowel. The amount of time spent with at least one forepaw on the bar
is determined. When the animal removes its paw, the time is recorded and the mouse is
repositioned on the bar. Four trials are conducted for each animal at 30, 60, 90, 120mins. An
animal is considered cataleptic if it remains on the bar for 60 seconds or more. The
percentage of cataleptic animals is calculated. The phenomenon of catalepsy predicts
antipsychotic activity as well as the potential of producing extrapyramidial side effects[21]
Antipsychotic induced weight gain
Mice were randomly divided into five groups. Each mouse was individually weighed before
administration of drugs. Hereafter, mice of groups I, II, III, IV and V received Normal saline,
Resperidone (control), Test drug 1, test drug 2 and combination of test drugs 1 and 2
respectively everyday for 4 weeks. Normal mice chow feed and water were provided as
usual. After 2 and 4 weeks, mice were individually weighed again. The difference in the mice
weight before the administrations of drugs, after 2 weeks and after 4 weeks of administrations
of drugs was then compared.
Forced Swim Test (FST)
In this test mice are forced to swim in a restricted space from which there is no escape and
become immobile. Individual mice were forced to swim in an open cylindrical container
containing 7 cm of water at 22.0 ±1.0ºC; the duration of immobility or struggling in a period
of 6 minutes was recorded. Immobility was evaluated as when mice ceased to struggle and
remained floating in the water, making only necessary movements necessary to keep its head
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above water. At the end of the session, the animal was removed from water and dried
gently.[23]
Mice of Group I, II, III, IV, V received Normal saline, Test drug 1, Test drug 2,
combination of test drugs 1 and 2 and standard drug (Imipramine 10mg/kg) respectively.
Tail suspension test
Mice of Group I, II, III, IV, V received Normal saline, Test drug 1, Test drug 2, combination
of test drugs 1 and 2 and standard drug (Imipramine 10mg/kg) respectively. The total
duration of immobility induced by tail suspension was measured according to the method
described by (Steru et al.). Mice were suspended on the edge of a table 50 cm above the floor
by the adhesive tape placed approximately 1 cm from the tip of the tail. The total duration of
immobility was recorded for a total 6 min.
Step down method
Mice of either sex were used for this test. A rectangular box 50×50×50cm with grid floor and
a movable platform in the center of the box is used. The grid floor is connected to a shock
device, which delivers scrambled foot shocks. Mice of groups II, III and IV received Test
drug 1, test drug 2 and combination of test drugs 1 and 2 respectively. Groups I and V
received Normal saline and standard drug(Piracetam 100mg/kg).
The actual experiment comprises of three stages. In the first stage of familiarization, the
animal is placed on the platform, which is released after raising the cylinder and a latency to
descend is measured. After 10 sec of exploration, it is returned to the home cage. During the
second stage of learning a foot shock (50Hz; 1.5mA; 1s) is administered as soon as the
animal descends from the platform and the animal is then returned to the home cage. In the
final stage of retention test, the animal is again placed on the platform after 24 hours of the
learning trial and the latency to step down is measured.
The test is concluded when the animal steps down or remains on the platform for a cut-off
time of 60 seconds. The average time of descent during the learning stage and during the
reaction test are measured.
Prolongation of the step-down learning is defined as latency. However, the variability of
results obtained by this method is very high and it is necessary to use a large number of
animals. The animals must be handled very gently and the foot-shock must be delivered
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immediately at the first contact of the animal with the floor. Further the room must be kept
sound proof during the entire procedure.[22]
Histopathological reports
The animals were euthanized using anesthetic ether and their brains were dissected out. The
isolated brains were stored in labeled containers containing neutral formalin (10% solution).
These brains were submitted to VIJAYA DIAGNOSTIC CENTER for further
histopathological studies.
RESULTS AND DISCUSSION
Table 1: Phytochemical Analysis of Ethanolic leaf extract of Nelumbo nucifera and
Eichhornia crassipes.
Constituent Nelumbo
nucifera
Eichhornia
crassipes
Alkaloids + +
Flavonoids + +
Steroids + +
Glycosides + +
Proteins + +
Phenolic compounds + +
Saponins - -
Triterpenoids - +
Carbohydrates + -
Tannins + -
(+) indicates presence; (-) indicates absence
Table 2: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Head Dip Test.
S.no Treatment Number of head dips
1 Normal saline 39.25±0.4787
2 Nelumbo nucifera 13.50±0.9574
3 Eichhornia crassipes 11.25±0.25
4 Nelumbo nucifera +
Eichhornia crassipes 8.5±0.2887
5 Standard
(Diazepam 2mg/kg) 13.75±0.8539
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The exploratory activity decreases significantly when doses of ethanolic leaf extract of
Nelumbo nucifera, Eichhornia crassipes and combination of both Nelumbo nucifera and
Eichhornia crassipes are given. The decrease effect is higher than that of mice treated with
Diazepam as standard drug.
Table 3: Effect of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Locomotor Activity.
S.no Treatment Locomotor activity in 10 mins
Before treatment After treatment Decrease %
1 Vehicle 96±21.33 96.75±20.66 -0.78%
2 Control
(Caffeine 40mg/kg) 91.25±4.049 160.3±7.016 -75.61%
3 Nelumbo nucifera 97.75±1.601 59.25±0.4787 39.38%
4 Eichhornia crassipes 132.8±6.074 69.25±0.4787 47.83%
5
Nelumbo nucifera +
Eichhornia crassipes 43±1.780 39.75±0.8539 7.55%
6 Standard
(Diazepam 3mg/kg) 101.5±1.190 85±1.080 16.25%
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Percentage decrease in locomotor activity was highest in mice treated with ethanolic extract
of leaves of Eichhornia crassipes. Ethanolic extracts of Nelumbo nucifera and and
combination of both Nelumbo nucifera and Eichhornia crassipes also showed high decrease
percentage in locomotor activity when compared to control and Standard drug Diazepam.
Table 4: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Rota-Rod Test
S.no Treatment Duration of stay on rota-rod
0 mins 30 mins 60 mins 120 mins
1 Vehicle 262.5±2.255 266.5±5.60 262.8±4.211 268±6.519
2 Nelumbo nucifera 267.3±6.725 312.8±12.10 226.8±13.58 260±5.788
3 Eichhornia crassipes 260±8.436 66±7.714 48.75±0.6296 193.8±2.136
4
Nelumbo nucifera +
Eichhornia crassipes 266±5.115 200.8±1.548 226.3±2.213 274±9.065
5 Standard
(Diazepam 10mg/kg) 267.5±3.862 55.75±1.493 44.50±1.555 105±2.483
The extract-treated mice were able to maintain their posture on the rotating rod without
falling for over 180 seconds, especially of those mice treated with Combination of both tests
drugs and that of Nelumbo nucifera and the cut-off time on the tread mill was used.
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Table 5: Effect of Ethanolic Extract of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Pentobarbitone Induced Sleep Test.
S.no Treatment Sleep time (Mins)
Onset of sleep Duration of sleep Prolongation %
1 Control
(Pentobarbitone) 21.25±1.25 44.25±1.548 ---
2 Nelumbo nucifera 17.5±1.443 52.75±5.662 19.20%
3 Eichhornia crassipes 8.25±0.6292 70.5±0.6455 59.32%
4
Nelumbo
nucifera+Eichhornia
crassipes
9.5±0.5 54.5±1.848 23.16%
5 Standard
(Diazepam 3mg/kg) 2.25±0.6292 110.3±4.090 149.15%
The ethanolic extract of Eichhornia crassipes and combination of bith test drugs significantly
shortened the sleep onset time (sleep latency) and prolonged sleeping time. They both also
significantly prolonged duration of sleep.
Table 6: Effect of Ethanolic Extract of Leaves of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination of Both Test Drugs (Test Drug 3) And
Standard Drug On Hole Cross Test.
S.no Treatment Number of movements (= number of hole cross)
0 min 30 min 60 min 90 min
1 Vehicle 6.75±0.75 7.5±0.2887 9±0.4782 7.5±0.2887
2 Nelumbo nucifera 5.5±0.2887 0.5±0.2887 3.5±0.2887 2.5±0.2887
3 Eichhornia crassipes 5±0.4082 1±0.0 5±0.4082 3.75±0.25
4
Nelumbo nucifera +
Eichhornia crassipes 5.75±0.8539 1.5±0.2887 5.5±0.2887 5.5±0.50
5
Standard
(Diazepam 1mg/kg) 4.75±0.4787 1.25±0.4787 1.5±0.6455 0.5±0.2887
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The ethanolic extracts of all three test drugs (Nelumbo nucifera, Eichhornia crassipes and
combination of Nelumbo nucifera and Eichhornia crassipes) significantly displayed a
suppression of motor activity and exploratory behavior in this test. The locomotor activity
lowering effect was evident in the second (30 min) and third observation (60 min) and
continued up to a fifth observation period (120 min).
Table 7: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Hot Plate Test.
S.no Treatment Reaction time in seconds
0 min 30 mins 60 mins 90 mins 150 mins
1 Vehicle 0.5±0.2887 1±0.0 0.5±0.2887 0.5±0.2887 1±0.0
2 Nelumbo nucifera 7±0.4082 7±0.8165 7±1.225 6.25±0.25 10±0.4082
3 Eichhornia crassipes 4±0.4082 7.5±0.2887 7.75±0.8539 5±1.080 8±0.8165
4 Nelumbo nucifera +
Eichhornia crassipes 7.75±0.4787 3.7±0.4787 16±0.7071 8±0.0 7.25±0.25
5
Standard
(Aspirin 10mg/kg) 5±0.4082 14.25±0.8539 14.25±0.8539 14.25±0.4787 15.75±1.493
All three doses of the ethanolic extracts of test drugs produced increase in latency time
when compared with the vehicle. At the third observation (60 mins), ethanolic extract of
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combination of Nelumbo nucifera and Eichhornia crassipes produced higher latency period
than the standard drug Aspirin.
Table 8: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Acetic Acid Induced Writhing Test.
S.no Treatment No. of writhings Inhibition %
1 Control
(Acetic acid 300mg/kg) 31.50±1.323 ---
2 Nelumbo nucifera 14.25±0.8539 54.76%
3 Eichhornia crassipes 16.25±2.056 48.41%
4 Nelumbo nucifera +
Eichhornia crassipes 6.25±0.8539 80.15%
5 Standard
(Ibuprofen 40mg/kg) 3.75±0.8539 88.09%
The administration of all three test drug extracts significantly inhibited writhing response
induced by acetic acid when compared to mice administered with Vehicle. Inhibition
percentage was found to be highest in combination of ethanolic extracts of Nelumbo nucifera
and Eichhornia crassipes.
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Table 9: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Tail Flick Test.
S.no Treatment Tail withdrawal time in seconds (Mean ± SEM)
15 mins 30 mins 60 mins 90 mins
1 Vehicle 0±0.0 0.25±0.25 1±0.4082 0.75±0.4787
2 Nelumbo nucifera 4.5±1.041 4.75±1.25 3.5±1.041 3.25±0.75
3 Eichhornia crassipes 3.25±0.8539 3.75±0.8539 2.75±0.4787 1.75±0.25
4 Nelumbo nucifera +
Eichhornia crassipes 4.75±0.8539 8.5±1.323 6.5±0.866 4.75±0.75
5 Standard
(Aspirin 100mg/kg) 3.75±0.4787 2.5±0.6455 2.5±0.2887 2±0.0
The tail withdrawal reflex time following administration of the ethanolic extracts of test
drugs was found to increase. The tail withdrawal time of mice administered Nelumbo
nucifera and combination of both drugs i.e, Nelumbo nucifera and Eichhornia crassipes was
observed to be higher than that of mice administered standard drug, Aspirin at all 4
observations (15 mins, 30 mins, 60 mins, 90 mins)
Table 10: effect of ethanolic extract of leaves of nelumbo nucifera (test drug 1),
eichhornia crassipes (test drug 2), combination of both test drugs (test drug 3) and
standard drug on isoniazid induced convulsion test.
S.no Treatment Onset of
seizures
Duration of
seizures
Number of
deaths Mortality %
1 Control
(Isoniazid 300mg/kg) 612±4.416 39.25±0.4787 4/4 100%
2 Nelumbo nucifera 651.3±1.315 35.75±2.287 2/4 50%
3 Eichhornia crassipes 697.8±1.702 33.25±1.797 1/4 25%
4 Nelumbo nucifera + 781.5±1.19 24.25±0.8539 0/4 0%
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Eichhornia crassipes
5 Standard
(Phenytoin 10mg/kg) 874±2.677 7.25±1.031 0/4 0%
Time of onset of seizures of all three drug administrated mice was observed to have increased
when compared to control. Duration of seizures steadily decreased when compared to control
as well. Highest increase in time ofonset of seizures was found in mice treated with
combination of ethanolic extracts Nelumbo nucifera and Eichhornia crassipes.
Table 11: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Elevated Plus Maze Test.
S.no Treatment
Number of entries in
open arms (Seconds)
(Mean ± SEM)
Time spent in open
arms (Seconds)
(Mean ± SEM)
1 Vehicle 1±0.0 36.25±8.004
2 Nelumbo nucifera 3.75±0.4787 126.8±2.287
3 Eichhornia crassipes 4.25±0.25 199.3±13.73
4 Nelumbo nucifera +
Eichhornia crassipes 4.5±1.5 215±31.32
5 Standard
(Diazepam 35mg/kg) 3.75±0.4787 234.3±5.105
Table 11 a:
S.no Treatment
Number of entries in
closed arms (Seconds)
(Mean ± SEM)
Time spent in closed
arms (Seconds)
(Mean ± SEM)
1 Vehicle 2.75±0.25 263.8±8.004
2 Nelumbo nucifera 4.75±0.4787 173.3±2.287
3 Eichhornia crassipes 3.75±0.9465 100.8±13.73
4 Nelumbo nucifera + 1.5±0.5 83.75±31.32
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Eichhornia crassipes
5 Standard
(Diazepam 35mg/kg) 1.25±0.25 65.75±5.105
The ethanolic extracts of Nelumbo nucifera and Eichhornia crassipes and combination of
both test drugs significantly increased the duration of stay of mice in open arm of apparatus
and decreased the duration of stay in the closed arm. Most time spent in open arm was
observed in mice administered combination of ethanolic extracts Nelumbo nucifera and
Eichhornia crassipes, whose readings are easily comparable with those of mice administered
standard drug, Diazepam.
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Table 12: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Novelty Induced Hypophagia.
S.no Treatment Home cage
(Mean±SEM)
Novel cage
(Mean±SEM)
1 Vehicle 62±3.582 302.3±4.385
2 Nelumbo nucifera 85.75±14.02 300±1.225
3 Eichhornia crassipes 62±4.708 38.25±8.3
4 Nelumbo nucifera +
Eichhornia crassipes 76±3.582 247±7.45
5 Standard
(Diazepam 10mg/kg) 63.5±1.323 82±5.148
Latency to drink the condensed milk mixture was highly significant in mice administered
ethanolic extracts of Eichhornia crassipes, which indicates high Anti-anxiety property of the
test drug. Latency time was observed to be lower than that of standard drug, Diazepam.
Table 13: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard.
Drug On Evation Test.
S.no Treatment
No. of remaining
mice in box after 5
mins
1 Vehicle 0
2 Nelumbo nucifera 2
3 Eichhornia crassipes 0
4 Nelumbo nucifera + Eichhornia
crassipes 3
5 Standard
(Diazepam 0.5mg/kg) 4
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The ethanolic extracts of Nelumbo nucifera and combination of Nelumbo nucifera and
Eichhornia crassipes produce a significant decrease in exploratory behaviour pattern as
evident from the results evasion tests. The number of mice remaining in the box were high
when compared to Vehicle and mice administered ethanolic extract of Eichhornia crassipes,
suggesting presence of CNS depressant property in Nelumbo nucifera and combination of
Nelumbo nucifera and Eichhornia crassipes test drugs.
Table 14: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Caffeine Induced Stereotypic Behavior Test.
S.no Treatment Stereotypic behavior score
5 mins 30 mins 60 mins
1 Control
(Caffeine 40mg/kg) 3±0.0 3±0.0 3±0.0
2 Nelumbo nucifera 2.5±0.2887 1±0.0 0.75±0.4787
3 Eichhornia crassipes 2.75±0.25 1.25±0.25 0.5±0.2887
4 Nelumbo nucifera +
Eichhornia crassipes 2.25±0.25 1.75±0.25 0.75±0.25
Table 14a.
S.no Treatment Locomotion score
5 mins 30 mins 60 mins
1 Control
(Caffeine 40mg/kg) 5±0.0 5±0.0 5±0.0
2 Nelumbo nucifera 5±0.0 4.25±0.25 1.5±0.2887
3 Eichhornia crassipes 5±0.0 4.25±0.4787 1.25±0.25
4 Nelumbo nucifera +
Eichhornia crassipes 4.75±0.25 4.25±0.4787 1.25±0.25
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Treatment of mice with ethanolic extracts of Nelumbo nucifera, Eichhornia crassipes and
combination of Nelumbo nucifera and Eichhornia crassipes significantly decreased
stereotypic behavior of mice with time, when compared to control.
Table 15: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard.
Drug On Catalepsy Test.
S.no Treatment Time spent on dowel (Seconds) (Mean±SEM)
30 mins 60 mins 90 mins 120 mins
1 Vehicle 26±0.4082 53.75±1.181 120.3±0.6292 100.3±0.25
2 Nelumbo nucifera 6.75±0.25 9.75±0.4787 4±0.0 5.25±0.6292
3 Eichhornia crassipes 11±0.4082 20±0.4082 17±0.4082 19.25±0.25
4 Nelumbo nucifera +
Eichhornia crassipes 6±0.0 20.75±0.25 9.25±0.4787 15±0.4082
5 Standard
(Resperidone 1mg/kg) 4±0.4082 7.25±0.4787 59.5±0.2887 73±1.581
Mice treated with ethanolic extracts of Nelumbo nucifera, Eichhornia crassipes and
combination of Nelumbo nucifera and Eichhornia crassipes had significantly decreased time
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spent on dowel at all four observation times (30 mins, 60 mins, 90 mins and 120 mins). Time
spent on dowel was lower than that of mice treated with Standard drug, resperidone,
suggesting excellent anti-psychotic property of test drugs, especially that of Nelumbo
nucifera.
Table 16: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Antipsychotic Induced Weight Gain
S.no Treatment Body weight in Grams
0 weeks 2 weeks 4 weeks
1 Vehicle 47.5±2.5 47.5±2.5 48.75±3.146
2 Control
(Risperidone 0.5mg/kg) 35±2.041 42.5±1.443 62.5±4.33
3 Nelumbo nucifera 40±4.082 45±2.041 45±2.041
4 Eichhornia crassipes 45±0.2887 42.50±4.330 40±3.536
5 Nelumbo nucifera +
Eichhornia crassipes 45±5.000 48.75±1.25 50±0.0
Weight gain of mice treated with ethanolic extracts of Nelumbo nucifera, Eichhornia
crassipes and combination of Nelumbo nucifera and Eichhornia crassipes at third
observation (4 weeks) was found to be minimal when compared to that of Control.
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Table 17: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Forced Swim Test.
S.no Treatment Immobility time in seconds
(Mean ± SEM)
1 Vehicle 283.5±12.9
2 Nelumbo nucifera 172±3.367
3 Eichhornia crassipes 213±5.066
4 Nelumbo nucifera + Eichhornia
crassipes 149±3.937
5 Standard (Imipramine 10mg/kg) 191.5±27.49
Immobility time in mice treated with with ethanolic extracts of Nelumbo nucifera, Eichhornia
crassipes and combination of Nelumbo nucifera and Eichhornia crassipes was observed to
have decreased when compared to control. Immobility time decrease was especially
significant in mice treated with ethanolic extracts of Nelumbo nucifera and combination of
Nelumbo nucifera and Eichhornia crassipes which were both lower than that of mice treated
with standard drug, Imipramine, suggesting excellent anti-depressant property of both.
Table 18: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Tail Suspension Test
S.no Treatment Immobility time in seconds
(Mean ± SEM)
1 Vehicle 122.8±3.010
2 Nelumbo nucifera 68±1.080
3 Eichhornia crassipes 46.56±1.033
4 Nelumbo nucifera + Eichhornia
crassipes 52.5±14.93
5 Standard (Imipramine 10mg/kg) 4±0.4082
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Immobility time in mice treated with with ethanolic extracts of Nelumbo nucifera, Eichhornia
crassipes and combination of Nelumbo nucifera and Eichhornia crassipes was observed to
have significantly decreased when compared to control, suggestive of anti-depressant
property of both extracts and their combination.
Table 19: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Step Down Test.
S.no Treatment Step down time in seconds
(Mean ± SEM)
1 Vehicle 82.5±9.845
2 Nelumbo nucifera 48.25±1.493
3 Eichhornia crassipes 147.3±1.377
4 Nelumbo nucifera +
Eichhornia crassipes 424.5±2.901
5 Standard
(Piracetam 10mg/kg) 402.8±2.626
Mice treated with ethanolic extract Eichhornia crassipes and combination of ethanolic
extracts of Nelumbo nucifera and Eichhornia crassipes were observed to have high latency to
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step down, when compared to mice treated with vehicle and Standard drug, piracetam.
Especially mice treated with combination of ethanolic extracts of Nelumbo nucifera and
Eichhornia crassipes were observed to have higher latency time to step down than that of
mice treated with standard drug, piracetam, suggestive of memory enhancing property of
combination of ethanolic extracts of Nelumbo nucifera and Eichhornia crassipes drug.
Table 22: Effect Of Ethanolic Extract Of Leaves Of Nelumbo Nucifera (Test Drug 1),
Eichhornia Crassipes (Test Drug 2), Combination Of Both Test Drugs (Test Drug 3)
And Standard Drug On Normothermic Animals.
S.no Treatment Temperature in ºC
0 min 30 mins 60 mins
1 Vehicle 36.7±0.1915 36.55±0.1658 36.43±0.04787
2 Nelumbo nucifera 36.75±0.1323 36.15±0.02887 35.78±0.1887
3 Eichhornia crassipes 36.58±0.1887 36.33±0.1652 36.4±0.1581
4 Nelumbo nucifera +
Eichhornia crassipes 37.18±0.2323 36.3±0.0 36.2±0.09129
Minimal or negligent change in body temperature in normothermic mice was observed in
mice treated with ethanolic extract Eichhornia crassipes and combination of ethanolic
extracts of Nelumbo nucifera and Eichhornia crassipes at all three observations (0 mins, 30
mins, 60 mins) suggesting drug safety and indicating that the drugs do not cause any
hyperthermicreactions.
HISTOPATHOLOGICAL RESULTS
Effect of Normal saline on Brain histopathology
Normal brain tissue was observed.
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Fig 1: Microphotograph of brain of mouse treated with Normal saline for one month
Effect of Nelumbo nucifera (ethanolic leaf extract) on Brain histopathology.
Neuronal degeneration was observed with mild oedema.
Fig 2: Microphotograph of brain of mouse treated with Nelumbo nucifera for one
month.
Effect of Eichhornia crassipes (Ethanolic leaf extract) on brain histopathology.
Neuronal degeneration and prominent glial reaction was observed.
Fig 3: Microphotograph of brain of mouse treated with Eichhornia crassipes for one
month
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Effect of combination of Nelumbo nucifera and Eichhornia crassipes (ethanolic leaf
extracts) on brain histopathology.
Edema of parenchyma with minimal neuronal degeneration was observed.
Fig 4: Microphotograph of brain of mouse treated with combination of Nelumbo
nucifera and Eichhornia crassipes for one month.
Effect of control (Mouse left untreated after induction of Psychoneurosis) on brain
histopathology.
Acute Neuronal degradation was observed.
Fig 5: Microphotograph of brain of mouse left untreated after psychoneurosis induction
for one month.
Histopathological reports suggest mild neuronal impairement in mice brain of Nelumbo
nucifera, Eichhornia crassipes and combinaton of Nelumbo nucifera and Eichhornia
crassipes treated mice, when compared to complete neuronal degradation and damage found
in brain of mouse left untreated after psychoneurosis induction. This suggests that the plant
extracts studied, have neuronal protective properties.
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4. CONCLUSION
Plant substances continue to serve as viable sources of drugs for the world population and
several plant-based products are in extensive clinical use (Roja & Rao, 2000). This study
paves the way for further attention and research to identify the active compounds
responsible for the plant neuropharmacological activity. Further studies should be undertaken
to elucidate the exact mechanism of action by which extracts exhert their neuropharmacological effect.
Awareness of local community must be enhanced incorporating the traditional knowledge with
scientific findings. Exploitation of the rapidly growing weed, Eichhornia crasipes can be done
on making the different pharmaceutical products, which will be widely available due to the
high availability and rapid growth. Hence, making the 'Best out of waste'.
ACKNOWLEDGEMENTS
This M.Pharmacy thesis is the result of a challenging journey, upon which many people have
contributed and given their support. I cannot thank my parents enough, for showing their
unlimited support and encouragement throughout the ups and downs of this research thesis.
My utmost, devout and sincerest gratitude and appreciation to Mrs. Mehnoor Farheen for
persevering with me as my advisor, guide and friend throughout the time it took me to
complete this research and write the dissertation. She was there for me from start to finish.
With great pleasure, I acknowledge my profound and sincere thanks to Principal Mrs.
Sunitha, Shadan Women 's College of Pharmacy, Hyderabad for constant encouragement
throughout the period, which helped me in the successful completion of this project. I wish to
express my gratitude to beloved Vice Principal Mrs. Nishath Farheen M.Pharm Ph.D, Shadan
Women's College of Pharmacy, Hyderabad for always finding time from her busy schedule to
answer queries and inspiring me to work my hardest. I must acknowledge as well the many
friends, lab assistants, and librarians who assisted, advised, and supported my research and
writing efforts. Last but not the least; i would like to thank our Lab helpers, Sultan and
Arshad, who were always flexible with their work timings, just to help me through my
research.
REFERENCES
1. H.U. Wittchen, F. Jacobi, J. Rehm, A. Gustavsson, M. Svensson, B. Jönsson, J. Olesen,
C. Allgulander, J. Alonso, C. Faravelli, L. Fratiglioni, P. Jennum, R. Lieb, A. Maercker,
J. van Os, M. Preisig, L. Salvador-Carulla, R. Simon and H.-C. Steinhausen, The size and
www.wjpps.com Vol 4, Issue 12, 2015.
903
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
burden of mental disorders and other disorders of the brain in Europe 2010, European
Neuropsychopharmacology., 2011; 21: 655–679.
2. Duke JA et al. Handbook of Medicinal Herbs, 2nd edn.CRC Press, 2002; 473.
3. Khare CP. Indian Herbal Remedies: Rational WesternTherapy, Ayurvedic, and Other
Traditional Usage,Botany, 1st edn. USA: Springer, 2004; 326–327.
4. Sridhar KR, Bhat R. Lotus: a potential nutraceuticalsource. J Agri Technol., 2007; 3:
143–155.
5. Chopra RN et al. Glossary of Indian Medicinal Plants.New Delhi: Council of Scientific
and IndustrialResearch, 1956; 174.
6. Liu CP et al. The extracts from Nelumbo nucifera suppress cell cycle progression,
cytokine genesexpression, and cell proliferation in human peripheralblood mononuclear
cells. Life Sci., 2004; 75: 699–716.
7. Kirtikar KR, Basu BD. Indian Medicinal Plants, 2nd edn.New Delhi International Book
Distributors, 1975; 116–120.
8. Chatterjee A, Pakrashi SC. The Treatise on IndianMedicinal Plants, vol. 1. New Delhi:
Publication andInformation Directorate, 1991; 94–96.
9. Onishi E et al. Comparative effects of crude drugs onserum lipids. Chem Pharm Bull.,
1984; 32: 646–650.
10. Mukherjee PK et al. A review of Nelumbo nucifera Gaertn. Ancient Sci Life., 1996; 15:
268–276.
11. Anon. Pharmacognosy of Indigenous Drugs, vol. 2. NewDelhi: Central Council for
Research in Ayurveda andSidhha, 1982; 806.
12. Varshney CK, Rzo´ska J. Aquatic weeds in South EastAsia, 1st edn. New Delhi:
Springer, 1976; 39.
13. Ling ZQ et al. Isolation, characterization, anddetermination of antioxidative activity of
oligomericprocyanidins from the seedpod of Nelumbo nucifera Gaertn. J Agriccult Food
Chem., 2005; 53: 2441–2445.
14. Chen Y et al. Separation, identification and rapiddetermination of liensine, isoliensinine
and neferinefrom embryo of the seed of Nelumbo nucifera Gaertn. byliquid
chromatography coupled to diode array detectorand tandem mass
spectrometry. J Pharm Biomed Anal., 2007; 43: 99–104.
15. Sanaa M. M. Shanab, Emad A. Shalaby, David A. Lightfoot, Hany A. El-Shemy,
Allelopathic Effects of Water Hyacinth [Eichhornia crassipes], PLOS DOI:
10.1371/journal.pone.0013200.
www.wjpps.com Vol 4, Issue 12, 2015.
904
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
16. Aravind R. Kurup, Divya Rajan, Dr. Jency Blesson, Sruthy Chandran, Thampatty A. R.,
Veena P. V., Detailed Analysis on Phytochemicals, Antioxidants, Antimicrobial Activity
of Eichhornia Crassipes, International Journal of Scientific Research, Volume : 2 | Issue :
2 | Feb 2013 • ISSN No 2277 – 8179.
17. Kennett GA, Dickinson SL and Curzon G. Central serotonergic responses and behavioral
adaptation to repeated immobilization: the effect of corticosterone synthesis inhibitor
metyrapone. Eur J Pharmacol., 1985; 119: 143–152.
18. Kennett GA, Dickinson SL, Curzon G. Enhancement of some 5-HT depressant behavioral
responses following repeated immobilization in rats. Brain Res., 1985; 330: 253–263.
19. Turner RA, Hebborn P. Screening Methods in Pharmacology. New York, NY, USA:
Academic Press., 1965.
20. Turner RA. Depressants of the central nervous system. In: Screening procedure in
Pharmacology. Vol. 1. 1st ed. New York: Academic press., 1972; 78‐88.
21. Costal B, Naylor RJ. On catalepsy and catatonia and the predictability of the catalepsy
test for neuroleptic activity. Psychopharmacol (Berl)., 1974; 34: 233-241.
22. Ahmad et al. Int J Pharm Pharm Sci, 5(4): 590-593 591.
23. Jarvik ME, Essmann WB. A simple one-trial learning stimulation in mice. Psychol Rep.,
1960; 6: 290.