1. R.SAI KALYANI
G.DAYA RANI
T.BHARGAVI
A.APARAJITHA
M.CHAITANYA
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2.  EPILEPSY:
Epilepsy is a collective term for a group of chronic seizure
disorder having in common ,sudden and transient episodes
(seizure) of loss or disturbance of consciousness , usually but
not always with a characteristic body movements (convulsions)
and sometimes with autonomic hyperactivity.
 SEIZURES :
Seizures are discrete, time limited alterations in brain function
including changes in motor activity, autonomic
function, consciousness or sensation that result from an abnormal
and excessive electrical discharge of a group of neurons with in the
brain.
 CONVULSIONS:
Convulsions are sudden, violent, involuntary contractions of
the muscles of the body, often accompanied by loss of
resconsciousness.
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3.  Psidium guajava (Linn.) belongs to the family-Myrtaceae has many
pharmaceutical applications.
 ROOTS - anti-emetic, and are useful in hemorrhages, diarrhoea and
dysentery especially in children, ulcers, gingivitis, proctoptosis and
vomiting.
 LEAVES -
wounds, ulcers, cholera, diarrhoea, vomiting, nephritis, cachexia and
epilepsy.
 FLOWERS - bronchitis, ophthalmodynia, colic andulemorrhagia.
 FRUITS - aphrodisiac, laxative, tonic and also useful in
diarrhoea, dysentery.9
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4. AIM :
An attempt is proposed to evaluate the effect of Psidium guajava
leaves extract on electro shock induced convulsions and
Pentylenetetrazole induced convulsions in rats or mice.
The whole study is divided into two phases:
1. Phase -1 : Identification and authentication of the plant Psidium
guajava(Linn.)
2. Phase -2 :To evaluate antiepileptic activity of hydro alcoholic extract
of Psidium guajava (Linn.) Leaves in various validated animal models
such as:
- PTZ (Pentylenetetrazole) Induced convulsive model
- Maximal Electro Shock (MES) Induced convulsive model
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5. METHODOLOGY:
The hydro alcoholic extract of the leaves was collected and the
following preliminary tests were performed:
Nature : Sticky
Color : Greenish
%Yield : 39.68(% w/w) in g.
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6.  PRELIMINARY PHYTOCHEMICAL TESTS:
EXPERIMENT OBSERVATION INFERENCE
Benedicts test Formation of orange red presence of reducing
precipitate sugar.
Fehling’s Test red precipitate presence of reducing sugar
Biuret test purplish violet color presence of proteins.
Millions test brick red precipitate Presence of proteins
Ninhydrin test purple or bluish color appears. Presence of amino acids
Tyrosine test dark red color Presence of amino acids
Mayer’s Test yellow cream precipitate presence of alkaloids
Dragendroff’s red precipitate presence of alkaloids
Test
Borntrager’s test Ammonical layer turns pink or red. Presence of glycosides
Lead acetate test Yellow colored precipitate Presence of flavonoids
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7. EXPERIMENT OBSERVATION INFERENCE
Salkowski’s Test lower layer turns red Sterol are present.
lower layer turns golden triterpenes are presents
yellow
Stain Test oily stain on filter paper presence of fixed oil
Lead acetate test White precipitate Presence of tannins and
phenols
Acetic acid test red color solution. Presence of tannins and
phenols
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8. Experimental animal : Albino mice of either sex weighing
between 20-30g were procured.
Principle of the test : Animals are dosed, one at a time, at 24 hour
intervals.
 The first animal receives a dose at the level of the best estimate
of the LD50.
 Depending on the outcome for the previous animal, the dose for
the next animal is adjusted up or down.
 If an animal survives, the dose for the next animal is increased;
if it dies, the dose for the next animal is decreased.
 After reaching the reversal of the initial outcome, i.e. the point
where an increasing (or decreasing) dose pattern is reversed by
giving a smaller (or a higher) dose, four additional animals are
dosed following the same UDP.
 The LD50 is calculated using the method of maximum likelihood.
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9. Procedure:
 The systemic acute oral toxicity (LD50) profile of the extract was
evaluated in female wistar albino rats according to OECD 425
guidelines.
 In brief, this method was carried out in three steps, the initial
investigation in which nine animals were used, three animals per
treatment group.
 The animals used were fasted overnight, note down the fasted body
weights and calculate the doses, the dose volume should not be
exceeded 1ml/100gm .
 The different doses selected were 500, 1000, 2000 mg/kg of the
extract per body weight.
 Animals are observed individually after dosing at least once during
the first 30 minutes, periodically during the first 24 hours, with
special attention given during the first 4 hours, and daily thereafter
for a total of 14 days.
 However, the duration of the observation period should not be fixed
rigidly. It should be determined by the toxic reactions, time of onset
and length of recovery period, and may thus be extended when
considered necessary.
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10. DETERMINATION OF ANTICONVULSANT
ACTIVITY :
PTZ INDUCED CONVULSIONS:
 Albino mice of either sex weighing between 22-25g were randomly
selected and segregated in to five groups, each group consisting of
six animals.
 Group A - Normal control (2%w/v Gum acacia p.o.)
 Group B - Standard (Diazepam 5mg/kg p.o)
 Group C - Leaf extract of Psidium guajava (Linn.)
(100mg/kg p.o)
 Group D - Leaf extract of Psidium guajava (Linn.)
(200 mg/kg p.o)
 Group E - Leaf extract of Psidium guajava (Linn.)
(400 mg/kg p.o)
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11. Experimental procedure(for anti convulsant activity)
 Group A served as normal control and was administered with 2%w/v Gum
acacia suspension orally, Group B with diazepam (5mg/kg p.o.) and served
as standard, Groups C, D and E with three different doses of leaf extracts
(low, medium and high respectively) hydroalcoholic of Psidium guajava(Linn.)
for seven consecutive days.
 On the eighth day one hour after the oral administration of either acacia
suspension/standard drug/extracts respectively to different groups, PTZ 60
mg/kg was administered subcutaneously.
 Each animal was then placed into individual plastic cages and were observed
initially for 30min and later up to 24 hrs.
The following parameters were recorded during test session of initial 30min
and up to 24 hrs respectively:
 ► Latency (onset of clonus)
 ► Onset of tonic-clonic convulsions
 ► Status of animal after 1hr
 ► Status of animal after 24 hrs
 ► Percent protection
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12. MAXIMAL ELECTRO SHOCK (MES) INDUCED
CONVULSIONS
Albino mice of either sex weighing between 22-25g were
divided into five groups each group was consisting of six
animals.
 Group A - Normal control (2% gum acacia p.o)
 Group B - Standard (Diazepam 5mg/kg p.o)
 Group C - Leaf extract of Psidium guajava (Linn.)
 (100mg/kg p.o)
 Group D - Leaf extract of Psidium guajava (Linn.)
(200 mg/kg p.o)
 Group E - Leaf extract of Psidium guajava (Linn.)
(400 mg/kg p.o)
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13. Experimental procedure(For MES induced
convulsions) :
 Group A served as control and was administered with 2% gum acacia
suspension,
 Group B with phenytoin (25mg/kg p.o.) and served as standard.
Group C, D and E with three different doses of hydroalcoholic
extracts of Psidium guajava(Linn.) (low, medium and high
respectively) for seven consecutive days.
 On the eighth day one hour after oral administration of acacia
suspension/standard drug/different extracts to respective groups,
MES seizures were induced by electroconvulsometer.
 A 60 mA current was delivered transauricularly for 0.2sec in mice..
 In the pilot study various phases of convulsions, viz., tonic flexion,
extension, clonus, stupor and mortality due to convulsions were
selected as the parameters.
 Phenytoin (25mg/kg p.o.) used as standard drug.
The following parameters were recorded during 1hr test session.
 Tonic flexion
 Tonic extension
 Clonus convulsions
 Percent protection
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14. RESULTS
. Assessment of anti-convulsant activity of hydro
alcoholic extract of psidium guajava leaves :
PTZ (Pentylenetetrazole) induced convulsions:
 Lower dose(100mg/kg)produced significant anticonvulsant activity
when compared to control
 Medium and high doses (200 & 400mg/kg respectively) exhibited a
very significant anti convulsant effect by increasing onset time of
seizures and reducing the duration of tonic-clonic seizures.
 The lower, medium and higher doses of leaves offered a protective
effect of 66.66%, 83.33% and 83.33% up to 1hr interval respectively.
 The standard drug diazepam (5mg/kg) exhibited a significant anti-
convulsant activity and offered 100% protection
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15. ONSET OF CONVULSIONS (seconds) STATUS OF
TREATEME ANIMAL AFTER
MEAN ± SEM
NT No. of animals in the group 1hr
Death/Recovery
1 2 3 4 5 6
Control
(2% Gum 654 648 699 687 643 692 670.5±10.135 6/0
acacia p.o.)
Diazepam
885 925 880 870 923 970 908.8±15.396** 0/6
(5mg/kg p.o.)
PGE
(100mg/kg 735 789 690 733 690 709 724.3±15.235* 2/4
p.o.)
PGE
(200mg/kg 790 755 820 805 782 795 806.1±15.844** 1/5
p.o.)
PGE
(400mg/kg 800 830 832 805 852 820 823.1±7.816** 1/5
p.o.)
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16. Effect of hydro alcoholic extract of psidium guajava(linn.) leaves
on MES induced convulsions (Duration of tonic extensor seizure) in mice.
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17. DURATION OF TONIC EXTENSOR
SEIZURES (seconds) MEAN ± SEM
No. of animals in the group
TREATEMENT
1 2 3 4 5 6
Control
(2% Gum 21 24 23 22 25 23 23 ±0.577
acacia p.o.)
Diazepam
7 5 6 4 8 5 5.83 ±0.600, **
(5mg/kg p.o.)
PGE 22.16
21 24 23 20 22 23
(100mg/kg p.o.) ±0.600
PGE
13 16 14 15 12 16 14.33 ±0.666, **
(200mg/kg p.o.)
PGE
10 14 11 13 12 11 12.16 ±0.600, **
(400mg/kg p.o.)
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18. ) Effect of hydro alcoholic extract of psidium guajava (linn.) leaves
on MES induced convulsions (Onset of clonic seizure) in mice
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19. DURATION OF
ONSET OF
TONIC
CLONUS
EXTENSOR %
AVG.WT. AVG.VOL.OF (Sec)
S.NO TREATEMENT (Sec) PROTECTION
(g) DOSE (ml)
(1hr)
Mean ± SEM
Mean ± SEM
Control
1 (2% Gum acacia 24.6 0.25 25±0.57 23 ±0.577 33.33
p.o.)
Diazepam (5mg/kg
2 23.8 0.23 6.8±0.60** 5.83 ±0.600** 100
p.o.)
PGE
3 23 0.23 24.1±0.60 22.16 ±0.600 33.33
(100mg/kg p.o.)
PGE
4 22 0.22 16.3±0.66** 14.33 ±0.666 ** 66.66
(200mg/kg p.o.)
PGE 83.33
5 24 0.24 13.8±0.60** 12.16 ±0.600 **
(400mg/kg p.o.)
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20. Effect of hydro alcoholic extract of psidium guajava (linn.) leaves
on MES induced convulsions in mice
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21.  The present study was carried out to find out,
evaluation of anti-epileptic activity on Psidium
Guajava (Linn.) leaves extract in mice.
 From the results obtained, we conclude that the
Psidium Guajava (Linn.) Leaves extract at higher
and medium doses produces highly significant
and sustained increase in the delay of onset of
convulsions and decrease in the no of
convulsions. This activity may be due to the
presence of different phytoconstituents viz,
flavanoids and saponins in the extract. However,
long term studies in different animals and
epileptic subjects may further substantial our
study result
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22.  Namara JO. Drug effective in the therapy of the Epilepsies. Goodman and
Gilman’s The Pharmacological basis of Therapeutics. 10th Ed. New York,
Mc Graw Hill; 2001: 521.
 Commission on Classification and Terminology of the International
League Against Epilepsy. Proposal for revised clinical and
electroencephalographic classification of epileptic seizures. Epilepsia,
1981, 22: 489-501
 HL Sharma, KK Sharma, Principles of pharmacology, Paras medical
publisher, Delhi, 2007; 1:531.
 http://en.wikipedia.org/wiki/Convulsion. Access date 25/12/2008
 Walczak TS, Leppik IE, D'Amelio M, Rarick J, Ahman P, Ruggles K, et al.
Incidence and risk factors in sudden unexpected death in epilepsy: a
prospective cohort study. Neurology 2001; 56: 519-525.
 Lathers C, Schraeder P. Epilepsy and Sudden Death. Dekker; 1990 NY.
 Hitiris N, Mohanraj R, Norrie J, Brodie M J. Mortality in epilepsy. Epilepsy
Behaviour; 2007, 363-376.
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23. THANK YOU
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