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Pharmacologyonline 3: 576-580 (2009) Pavan et al
576
PHYTOCHEMICAL SCREENING ANTIEPILEPTIC& ANALGESIC ACTIVITY
OF LEAF EXTRACT OF PASSIFLORA FOETIDA
Pavan Kumar Reddy K.,1
Bhagavan Raju M., 1
Sreedevi P., 1
Pranali Pandit 2
Veena Rani I., 3
Veena G.,3
1
Department of Pharmaceutical Chemistry, CM College of Pharmacy, Maisammaguda,
Dhulapally Hakimpet post, Kompally, Sec bad-500 014, Andhra Pradesh, India
2
Department of Pharmacognosy , CM College of Pharmacy, Maisammaguda, Dhulapally
Hakimpet post, Kompally, Sec bad-500 014, Andhra Pradesh, India
3
Department of Pharmacology , CM College of Pharmacy, Maisammaguda, Dhulapally
Hakimpet post, Kompally, Sec bad-500 014, Andhra Pradesh, India
Summary
Antiepileptic activity of passiflora foetida was evaluated in mice against maximum
electroshock induced convulsions and pentylenetetrazole induced covulsions.Screening
of analgesic activity was evaluated using Eddy’s hot plate method using aspirin as
standard.Antispamodic activity was evaluated in mice using Diazepam as standard.
Hypnotic activity was evaluated using thiopentone as standard.
Key words: Passiflora foetida, Antiepileptic activity, Analgesic activity
Introduction
Passiflora foetida is a perennial creeping vine which has an edible fruit and
leaves that have a mild rank aroma. Literature study reveals it has been used traditionally
in various countries for treatment of stress, anxiety, depression, insomnia, cardiac
arrhythmia and tension related asthma (1-2). Passiflora edulis sims possesses
antibacterial, cytotoxic and anti-oxidant activity (3-4).Leshmanicidal activity of
passifloricin A and derivatives were reported from the resin extract of passiflora foetida
(5).Benzoflavone moiety present in passiflora incarnate were responsible for antiaxiety
activity(6).Several HPLC methods has been employed for qualitative and quantitative
analysis of C-glycosylflavones in P.incarnata(7).The active constituents of Passiflora
foetida are hydrocyanic acid, groups of flavone alkaloids, harman alkaloids ,alkaloids,
phenols, cyanogenic compounds , passifloricins,polyketides and alpha-pyrones( 8-
10).The present study was undertaken to investigate antiepileptic and analgesic activity of
methanol extract of passiflora foetida.
Pharmacologyonline 3: 576-580 (2009) Pavan et al
577
Materials and methods
Dried leaves of Passiflora foetida were obtained from Kotla vijaybhaskar reddy
botanical garden, Hyderabad Andhra Pradesh, India and authenticated by Pranali Pandit,
pharmacognist, CM College of pharmacy, Hyderabad. A voucher specimen has been
deposited at the museum of our college. The leaves of passiflora foetida were air-dried.
The leaves were subjected to soxhlet extraction using different methanol for 48 hrs.The
extract was concentrated using rotary vacuum to get the solid mass. The phytochemical
investigation of various extracts are represented in Table-1, yield color and consistency
of various extracts are represented in Table-2
TABLE-1
Extract Constituents investigated
Petroleum ether Carbohydrates , flavanoids
Chloroform Carbohydrates,alkaloids,flavanoids,glycosides
Methanol Carbohydrates,tannins,triterpenes,flavanoids,glycosides
Hydroalcoholic Saponins,glycosides,alkaloids,carbohydrates,triterpenes
TABLE-2
Extract Yield(% W /W) Co lour consistency
Petroleum ether 5.0 Reddish brown Hard waxy
Chloroform 3.0 Green Waxy
Methanol 20 Dark brown Resinous
Hydro alcoholic 13.2 Reddish brown Resinous
Antiepileptic activity
Swiss albino mice male weighing 20-25 g were housed in groups of six under
standard lab conditions ( temperature 250
C ±10
C, relative humidity 55 ± 5% , 12h:12h
dark: light cycle) with standard pellet food and water ad libitum.The animals were
transferred to the laboratory atleast 1 hr before the start of the experiment. The
experiments were performed during the day and as per the guidelines of the Committee
for the purpose of supervision and control of Experiments on Animals (CPSCEA),
Government of India.
Maximum electroshock (MES) –induced convulsions
Mice were divided into 5 groups (n=6).Group I served as control ad received only
vehicle. Groups II, III and IV were treated orally with PG at a dose 30, 100 and 300
mg/kg body weight. Group V received 20 mg/Kg i.p phenytoin.The mice received a
current of 50mA for 0.2 sec duration through electroconvulsiometer using ear electrodes
after 60 min of oral administration of PG and 30 min after i.p administration of
phenytoin.The incidence and duration of extensor tonus was noted. A complete abolition
of hind limb tonic extension was considered as 100% protection (9). The results were
reported in Table-2.
Pharmacologyonline 3: 576-580 (2009) Pavan et al
578
Pentylenetetrazol (PTZ)-induced convulsions
Five groups of mice ( n=6) were treated with 80mg/kg s.c PTZ, 60 min after oral
administration of either PG ( 30, 100 and 300 mg/kg) or vehicle or 30 min after i.p
administration of diazepam ( 4mg/kg).The animals were observed for 30 min for onset,
presence or absence of clonic convulsions and mortality(9). The results were reported in
Table-3.
TABLE-2
Effect of methanol extract and phenytoin against MES-induced convulsions
Groups Duration of hind limb
Extension ( Sec)
Mean±SEM
n=6 Death P
( Dunn’s test)
Control 13.3±0.2 5/6
Passiflora methanol extract
( 30mg/kg.b.wt)
12.5±0.6 3/6 >0.05
Passiflora methanol extract
( 100mg/kg.b.wt)
5.0±0.4 0/6 <0.01
Passiflora methanol
extract(300mg/kg.b.wt)
5.2±0.2 0/6 <0.01
Phenytoin 4.0±1.0 0/6 <0.01
The data were analyzed by kruskal-Wallis test KW =22.26 (non parametric
ANOVA, P=0.0002, from chi square distribution) followed by Dunn’s multiple
comparison test.
TABLE-3
Effect of methanol extract and Diazepam against PTZ-Induced convulsions
Groups Onset of myoclonic
spasm( sec)
Mean±SEM
Onset of clonic
convulsions(sec)
Mean±SEM
Death P
n=6
Control 68.25 ±0.2 105±0.6 5/6
Passiflora methanol
extract ( 30mg/kg.b.wt)
119±12.8 164±20.0 4/6
Passiflora methanol
extract ( 100mg/kg.b.wt)
254±1.2 220±22.0 5/6
Passiflora methanol
extract(300mg/kg.b.wt)
A A 0/6
Diazepam A A 0/6
The data were analyzed by krushkal-Wallis test KW= 14.126( non parametric
ANOVA, P= 0.0005, from chi square distribution) followed by Dunn’s multiple
comparison test P. 0.05 ( Not significant) p< 0.001( Highly significant) A-Absence of
any seizure activity.
Pharmacologyonline 3: 576-580 (2009) Pavan et al
579
Analgesic activity
Screening of analgesic activity was carried in animal models which showed
reaction time 3-5 seconds (10). By using Eddy’s hot plate method screening of analgesic
activity is carried, the animals used are albino mice of either sex weighing 25-30 gm.
Mice were divided into 4 groups of sex and tested for 4 hours. Group I received Tween-
80 1% i.p and served as control. Group II received Analgin and served as standard. Group
III & IV received the methanol extract of passiflora foetida 50 mg/kg and 100mg/kg
(i.p).The observations were made at 30 min interval upto 4 hours. The results are shown
in Table-4
TABLE-4
Analgesic activity of methanol extract of passiflora foetida
Group Reaction time in seconds
0 min 30 min 60 min 120 min 180 min 240 min
I 5.96±0.1 5.56±0.0 3.51±0.21 3.24±0.10 3.62±0.24 3.82±0.30
II 5.48±0.1 14.25±0.2 15.11±0.5 16.50±0.2 17.20±0.2 5.5±0.92
III 5.10±0.1 11.40±0.1 12.26. ±0.1 14.87±2.6 16.87±1.0 7.9±0.2
IV 5.50±0.1 5.84±0.6 5.12±0.2 3.2±0.1 5.62±0.2 4.93±1.2
P<0.001 vs. control student t test
Results and Discussion
Antiepileptic activity was evaluated using Maximum electricshock induced
convulsions and pentylenetetrazol-induced convulsions .Doses at 30 mg/kg body wt did
not show significant anticonvulsant activity. Doses at 100 and 300 mg/kg significantly
reduced the severity of seizures when compared to standard drug phenytoin, in MES
induced convulsions, but in the case of pentylenetetrazole induced convulsions, the
convulsions were completely abolished at doses 100mg/kg & 300 mg/kg b.wt when
compared to standard drug diazepam. The methanol extract of leaves of passiflora foetida
showed antiepileptic activity in a dose dependant manner. The detailed pharmacological
activity have to be investigated whether it is due to inhibiting Na+ channels/Ca++
channels, GABA/Beznzodiazepine, NMDA receptor complex. The extract of passiflora
foetida may be effective in generalized tonic seizures and absence seizures. Detailed EEG
(Electroencephalogram) discharge studies for different convulsions whether it due to
imbalance between which excitatory and inhibitory influences in the brain should be
studied for the extracts of passiflora foetida.
Analgesic activity was evaluated using Eddy’s hot plate method. The methanol
extract of passiflora foetida exhibited maximum analgesic activity at 120 min.In the
present study the methanol extract showed good analgesic activity at 100mg/kg when
compared to 50mg/kg. The mode of action of analgesics is on peripheral or central
nervous system. Any damage to tissue or injury leads to pain and inflammation .opiod
analgesics produce analgesia by binding to specific G protein coupled receptors that are
located in brain and spinal cord regions involved in the transmission and modulation of
pain.
Pharmacologyonline 3: 576-580 (2009) Pavan et al
580
Peripherally acting analgesics act by blocking the generation of impulses at
chemoreceptor pain site, but centrally acting analgesics not only raise the threshold for
pain but also alter the physiologic response to pain and suppress the patient’s anxiety and
apprehension. The pharmacological action of extract may be peripherally. The detailed
investigation of mechanism of action of drug should be studied. The isolation of active
principle from the crude drug responsible for anticonvulsant and antiepileptic activity
should be studied.
References
1.Soulimani R, Younos C, jarmouni S, Bousta D, Misslin R and Mortier F.Behavioural
effects of Passiflora incarnate L and its indole alkaloid and flavanoid derivatives and
maltol in the mouse.J.Ehnopharmacol 1997;57:11-20.
2. Abourshed EA, Vander plank JR and Khan IA. High-speed extraction and HPLC
finger print of medicinal plants-I. Application to Passiflora flavanoids.Pharm.Biol 2002;
40: 81-91.
4.Farhana AR, Mahmuda H, Laizuman N, Monirul Islam Md.Antibacterial, cytotoxic and
Antioxidant activity of Passiflora Edulis Sims. European journal of scientific research
2009; 31:92-598.
5. Wilson CG, Winston QF and Fernando EL.Leshmanicidal activity of Passifloricin A
and derivatives. Molecules 2004; 9:666-672.
6. Dhawan K, Kumar S and Sharma A.Anti-anxiety studies on extracts of Passiflora
incarnate Linneus. J.Ethnopharmacol 2001; 78:165-170.
7. Rehwald A, Meier B and Sticher O.Qualitative and quantitative reversed phase high
performance liquid chromatography of flavanoids in passiflora incarnata L. Pharm Acta
Helv 1994; 69:153-158.
8.Echeverri F, Arango V, Quinones W, Torres F, Escobar G, Rosero Y, Archbold
R.Passifloricins,Polyketides alpha-pyrones from Passiflora foetida resin.Phytochemistry
2001;56:881-885.
9. Vogel GH, Vogel WH. Drug discovery and evaluation. Pharmacological assays.Berlin:
Springer-Verlag: 1997.
10. Turner RA.Screening methods in pharmacology. New York: Academic press; 1965;
158.

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Phytochemical screening antiepileptic and analgesic activity of leaf extract of passiflora foetida.pdf

  • 1. Pharmacologyonline 3: 576-580 (2009) Pavan et al 576 PHYTOCHEMICAL SCREENING ANTIEPILEPTIC& ANALGESIC ACTIVITY OF LEAF EXTRACT OF PASSIFLORA FOETIDA Pavan Kumar Reddy K.,1 Bhagavan Raju M., 1 Sreedevi P., 1 Pranali Pandit 2 Veena Rani I., 3 Veena G.,3 1 Department of Pharmaceutical Chemistry, CM College of Pharmacy, Maisammaguda, Dhulapally Hakimpet post, Kompally, Sec bad-500 014, Andhra Pradesh, India 2 Department of Pharmacognosy , CM College of Pharmacy, Maisammaguda, Dhulapally Hakimpet post, Kompally, Sec bad-500 014, Andhra Pradesh, India 3 Department of Pharmacology , CM College of Pharmacy, Maisammaguda, Dhulapally Hakimpet post, Kompally, Sec bad-500 014, Andhra Pradesh, India Summary Antiepileptic activity of passiflora foetida was evaluated in mice against maximum electroshock induced convulsions and pentylenetetrazole induced covulsions.Screening of analgesic activity was evaluated using Eddy’s hot plate method using aspirin as standard.Antispamodic activity was evaluated in mice using Diazepam as standard. Hypnotic activity was evaluated using thiopentone as standard. Key words: Passiflora foetida, Antiepileptic activity, Analgesic activity Introduction Passiflora foetida is a perennial creeping vine which has an edible fruit and leaves that have a mild rank aroma. Literature study reveals it has been used traditionally in various countries for treatment of stress, anxiety, depression, insomnia, cardiac arrhythmia and tension related asthma (1-2). Passiflora edulis sims possesses antibacterial, cytotoxic and anti-oxidant activity (3-4).Leshmanicidal activity of passifloricin A and derivatives were reported from the resin extract of passiflora foetida (5).Benzoflavone moiety present in passiflora incarnate were responsible for antiaxiety activity(6).Several HPLC methods has been employed for qualitative and quantitative analysis of C-glycosylflavones in P.incarnata(7).The active constituents of Passiflora foetida are hydrocyanic acid, groups of flavone alkaloids, harman alkaloids ,alkaloids, phenols, cyanogenic compounds , passifloricins,polyketides and alpha-pyrones( 8- 10).The present study was undertaken to investigate antiepileptic and analgesic activity of methanol extract of passiflora foetida.
  • 2. Pharmacologyonline 3: 576-580 (2009) Pavan et al 577 Materials and methods Dried leaves of Passiflora foetida were obtained from Kotla vijaybhaskar reddy botanical garden, Hyderabad Andhra Pradesh, India and authenticated by Pranali Pandit, pharmacognist, CM College of pharmacy, Hyderabad. A voucher specimen has been deposited at the museum of our college. The leaves of passiflora foetida were air-dried. The leaves were subjected to soxhlet extraction using different methanol for 48 hrs.The extract was concentrated using rotary vacuum to get the solid mass. The phytochemical investigation of various extracts are represented in Table-1, yield color and consistency of various extracts are represented in Table-2 TABLE-1 Extract Constituents investigated Petroleum ether Carbohydrates , flavanoids Chloroform Carbohydrates,alkaloids,flavanoids,glycosides Methanol Carbohydrates,tannins,triterpenes,flavanoids,glycosides Hydroalcoholic Saponins,glycosides,alkaloids,carbohydrates,triterpenes TABLE-2 Extract Yield(% W /W) Co lour consistency Petroleum ether 5.0 Reddish brown Hard waxy Chloroform 3.0 Green Waxy Methanol 20 Dark brown Resinous Hydro alcoholic 13.2 Reddish brown Resinous Antiepileptic activity Swiss albino mice male weighing 20-25 g were housed in groups of six under standard lab conditions ( temperature 250 C ±10 C, relative humidity 55 ± 5% , 12h:12h dark: light cycle) with standard pellet food and water ad libitum.The animals were transferred to the laboratory atleast 1 hr before the start of the experiment. The experiments were performed during the day and as per the guidelines of the Committee for the purpose of supervision and control of Experiments on Animals (CPSCEA), Government of India. Maximum electroshock (MES) –induced convulsions Mice were divided into 5 groups (n=6).Group I served as control ad received only vehicle. Groups II, III and IV were treated orally with PG at a dose 30, 100 and 300 mg/kg body weight. Group V received 20 mg/Kg i.p phenytoin.The mice received a current of 50mA for 0.2 sec duration through electroconvulsiometer using ear electrodes after 60 min of oral administration of PG and 30 min after i.p administration of phenytoin.The incidence and duration of extensor tonus was noted. A complete abolition of hind limb tonic extension was considered as 100% protection (9). The results were reported in Table-2.
  • 3. Pharmacologyonline 3: 576-580 (2009) Pavan et al 578 Pentylenetetrazol (PTZ)-induced convulsions Five groups of mice ( n=6) were treated with 80mg/kg s.c PTZ, 60 min after oral administration of either PG ( 30, 100 and 300 mg/kg) or vehicle or 30 min after i.p administration of diazepam ( 4mg/kg).The animals were observed for 30 min for onset, presence or absence of clonic convulsions and mortality(9). The results were reported in Table-3. TABLE-2 Effect of methanol extract and phenytoin against MES-induced convulsions Groups Duration of hind limb Extension ( Sec) Mean±SEM n=6 Death P ( Dunn’s test) Control 13.3±0.2 5/6 Passiflora methanol extract ( 30mg/kg.b.wt) 12.5±0.6 3/6 >0.05 Passiflora methanol extract ( 100mg/kg.b.wt) 5.0±0.4 0/6 <0.01 Passiflora methanol extract(300mg/kg.b.wt) 5.2±0.2 0/6 <0.01 Phenytoin 4.0±1.0 0/6 <0.01 The data were analyzed by kruskal-Wallis test KW =22.26 (non parametric ANOVA, P=0.0002, from chi square distribution) followed by Dunn’s multiple comparison test. TABLE-3 Effect of methanol extract and Diazepam against PTZ-Induced convulsions Groups Onset of myoclonic spasm( sec) Mean±SEM Onset of clonic convulsions(sec) Mean±SEM Death P n=6 Control 68.25 ±0.2 105±0.6 5/6 Passiflora methanol extract ( 30mg/kg.b.wt) 119±12.8 164±20.0 4/6 Passiflora methanol extract ( 100mg/kg.b.wt) 254±1.2 220±22.0 5/6 Passiflora methanol extract(300mg/kg.b.wt) A A 0/6 Diazepam A A 0/6 The data were analyzed by krushkal-Wallis test KW= 14.126( non parametric ANOVA, P= 0.0005, from chi square distribution) followed by Dunn’s multiple comparison test P. 0.05 ( Not significant) p< 0.001( Highly significant) A-Absence of any seizure activity.
  • 4. Pharmacologyonline 3: 576-580 (2009) Pavan et al 579 Analgesic activity Screening of analgesic activity was carried in animal models which showed reaction time 3-5 seconds (10). By using Eddy’s hot plate method screening of analgesic activity is carried, the animals used are albino mice of either sex weighing 25-30 gm. Mice were divided into 4 groups of sex and tested for 4 hours. Group I received Tween- 80 1% i.p and served as control. Group II received Analgin and served as standard. Group III & IV received the methanol extract of passiflora foetida 50 mg/kg and 100mg/kg (i.p).The observations were made at 30 min interval upto 4 hours. The results are shown in Table-4 TABLE-4 Analgesic activity of methanol extract of passiflora foetida Group Reaction time in seconds 0 min 30 min 60 min 120 min 180 min 240 min I 5.96±0.1 5.56±0.0 3.51±0.21 3.24±0.10 3.62±0.24 3.82±0.30 II 5.48±0.1 14.25±0.2 15.11±0.5 16.50±0.2 17.20±0.2 5.5±0.92 III 5.10±0.1 11.40±0.1 12.26. ±0.1 14.87±2.6 16.87±1.0 7.9±0.2 IV 5.50±0.1 5.84±0.6 5.12±0.2 3.2±0.1 5.62±0.2 4.93±1.2 P<0.001 vs. control student t test Results and Discussion Antiepileptic activity was evaluated using Maximum electricshock induced convulsions and pentylenetetrazol-induced convulsions .Doses at 30 mg/kg body wt did not show significant anticonvulsant activity. Doses at 100 and 300 mg/kg significantly reduced the severity of seizures when compared to standard drug phenytoin, in MES induced convulsions, but in the case of pentylenetetrazole induced convulsions, the convulsions were completely abolished at doses 100mg/kg & 300 mg/kg b.wt when compared to standard drug diazepam. The methanol extract of leaves of passiflora foetida showed antiepileptic activity in a dose dependant manner. The detailed pharmacological activity have to be investigated whether it is due to inhibiting Na+ channels/Ca++ channels, GABA/Beznzodiazepine, NMDA receptor complex. The extract of passiflora foetida may be effective in generalized tonic seizures and absence seizures. Detailed EEG (Electroencephalogram) discharge studies for different convulsions whether it due to imbalance between which excitatory and inhibitory influences in the brain should be studied for the extracts of passiflora foetida. Analgesic activity was evaluated using Eddy’s hot plate method. The methanol extract of passiflora foetida exhibited maximum analgesic activity at 120 min.In the present study the methanol extract showed good analgesic activity at 100mg/kg when compared to 50mg/kg. The mode of action of analgesics is on peripheral or central nervous system. Any damage to tissue or injury leads to pain and inflammation .opiod analgesics produce analgesia by binding to specific G protein coupled receptors that are located in brain and spinal cord regions involved in the transmission and modulation of pain.
  • 5. Pharmacologyonline 3: 576-580 (2009) Pavan et al 580 Peripherally acting analgesics act by blocking the generation of impulses at chemoreceptor pain site, but centrally acting analgesics not only raise the threshold for pain but also alter the physiologic response to pain and suppress the patient’s anxiety and apprehension. The pharmacological action of extract may be peripherally. The detailed investigation of mechanism of action of drug should be studied. The isolation of active principle from the crude drug responsible for anticonvulsant and antiepileptic activity should be studied. References 1.Soulimani R, Younos C, jarmouni S, Bousta D, Misslin R and Mortier F.Behavioural effects of Passiflora incarnate L and its indole alkaloid and flavanoid derivatives and maltol in the mouse.J.Ehnopharmacol 1997;57:11-20. 2. Abourshed EA, Vander plank JR and Khan IA. High-speed extraction and HPLC finger print of medicinal plants-I. Application to Passiflora flavanoids.Pharm.Biol 2002; 40: 81-91. 4.Farhana AR, Mahmuda H, Laizuman N, Monirul Islam Md.Antibacterial, cytotoxic and Antioxidant activity of Passiflora Edulis Sims. European journal of scientific research 2009; 31:92-598. 5. Wilson CG, Winston QF and Fernando EL.Leshmanicidal activity of Passifloricin A and derivatives. Molecules 2004; 9:666-672. 6. Dhawan K, Kumar S and Sharma A.Anti-anxiety studies on extracts of Passiflora incarnate Linneus. J.Ethnopharmacol 2001; 78:165-170. 7. Rehwald A, Meier B and Sticher O.Qualitative and quantitative reversed phase high performance liquid chromatography of flavanoids in passiflora incarnata L. Pharm Acta Helv 1994; 69:153-158. 8.Echeverri F, Arango V, Quinones W, Torres F, Escobar G, Rosero Y, Archbold R.Passifloricins,Polyketides alpha-pyrones from Passiflora foetida resin.Phytochemistry 2001;56:881-885. 9. Vogel GH, Vogel WH. Drug discovery and evaluation. Pharmacological assays.Berlin: Springer-Verlag: 1997. 10. Turner RA.Screening methods in pharmacology. New York: Academic press; 1965; 158.