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Corresponding author
N.M.A. Rasheed
rasheed_chem@yahoo.co.in
Please cite this article in press as N.M.A. Rasheed et al. Phytochemical evaluation and anthelmintic activity of ethanolic leaves
extract of Passiflora foetida Linn. Indo American Journal of Pharm Research.2013:3(7).
Copy right 2013 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
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Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly
cited.
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Vol 3, Issue 7, 2013. N.M.A. Rasheed et al. ISSN NO: 2231-6876
Introduction
Passiflora foetida Linn also called as stinking passionflower or wild water lemon belongs to Passifloraceae
family. It is a fetid, herbaceous, hairy, perennial vine, scrambling or climbing to 5m or more by axillary,
unbranched, coiling tendrils with soft to hard, yellow to brown hairs, distributed and found wild in several parts
of India. It is native to the Southwestern United States (Southern Texas and Arizona), Mexico, the Caribbean,
Central America, and much of South America. It has been introduced to tropical regions around the world, such
as Southeast Asia and Hawaii[1,2]. The stems are thin and wiry, covered with minute sticky yellow hairs. Older
stems become woody. The leaves are alternate, palmately three-five lobed and viscid-hairy. The flowers are
white to pale cream coloured, about 5-6 cm in diameter and with an epicalyx of pinnatifid bracteoles. The fruit
is globose up to 2.5 cm in diameter, hairy, yellowish when ripe and has numerous black seeds embedded in the
pulp[3]. Leaves have a mild aroma and ripened fruit is edible. The plant is reported to contain alkaloids,
phenols, glycosides, flavonoids, cyanogenic compounds, passifloricins, polypeptides and alpha-pyrones.[4]
Passiflora foetida Linn. reported to possess sedative, hypnotic, antispasmodic and anodyne properties[5]. Tea of
its leaves is used as an expectorant and for nervous disorders. Traditionally it is used for diarrhoea, intestinal
tract, throat, ear infections, fever and skin diseases. It is also used in eczema, inflammation and pain[1,2].
Literature study shows that the fruits of Passiflora foetida Linn. possess hepatoprotective activity may be
attributed to flavonoids present in the fruits of Passiflora foetida Linn[6]. The antibacterial properties of leaf
and fruit (ethanol and acetone) extracts were screened against four human pathogenic bacteria i.e. Pseudomonas
putida, Vibrio cholerae, Shigella flexneri and Streptococcus pyogenes by well-in agar method. The results
showed the leaf extract having remarkable activity against all bacterial pathogens compared to fruits[7]. Besides
these, the amount of harmaline, a betacarboline alkaloid in Passiflora foetida was estimated by comparing the
peak area of standard and that present in the leaf extract. The harmaline content present in the extract was
estimated to be 0.75% w/w[8]. Antinociceptive, antidiarrhoeal and cytotoxic activities of ethanolic extract of
whole plant were also studied. The extract produced significant (P<0.001) writhing inhibition in acetic acid-
induced writhing in mice. The extract also showed antidiarrhoeal activity on castor oil induced diarrhoea in
mice and cytotoxic activity against brine shrimp Artemia salina.[9] There are no reports on systematic and
scientific study of anthelmintic activity of leaf extract for passiflora foetida Linn. In the present study, we report
the anthelmintic activity of ethanolic extract of the leaves of Passiflora foetida Linn.
of approximately equal size. They were collected from local place, washed and kept in water.
Drugs
Ethanolic extract of Passiflora foetida Linn. was tested in various doses in each group. Normal saline water was
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used as control. Albendazole was used as the standard drug for the study with ethanolic extract.
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alkaloids.
Wagners test: 2 ml of the extract and 0.2 ml of dilute hydrochloric acid were placed in a test tube. Then 1 ml of
iodine solution (Wagners reagent) was added. Formation of brown/reddish precipitate indicates the presence of
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alkaloids.
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Hagers test: 2 ml solution of the extract and 0.2 ml of dilute hydrochloric acid were placed in a test tube. Then
1 ml of picric acid solution (Hagers reagent) was added. formation of yellow coloured precipitate indicates the
presence of alkaloids.
HPTLC Analysis:
Preparation of ethanolic Extract
Five grams powder of finely powdered leaves were dissolved in 100 ml of ethanol using a soxhlet
extraction and the extract was filtered through Whattmann No. 41 filter paper and concentrated over the water
bath to 20 ml. The solution obtained was used as sample for the determination of components.
The sample applied as a band of 10mm with the help of Automatic TLC applicator system of the
DESAGA Sarstedt Gruppe on Precoated Aluminium Sheets of Silica Gel 60 F254 (Merck). After trying with
various solvent systems with variable volume ratios, the suitable solvent system as stated above is selected in its
proportional ratio and developed in the Twin through chamber of TLC to the maximum height of the plate so
that it can be able to separate the components on the polar phase of silica gel and that of mobile phase of solvent
system[14-16].
with the Densitometer CD60 of DESAGA Sarstedt Gruppe system under the UV range of 366nm appearing a
maximum number of components. A corresponding densitograms was obtained as shown in the figures 2a in
which peaks are appeared for the corresponding spots being detected in the densitometer while scanning and the
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peaks area under the curve corresponds to the concentration of the component in the sample for the
concentration we applied on the TLC plate.
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Pharmacological Study
Anthelmintic Activity
Nargund[17] method was followed for the screening of anthelmintic activity on adult Indian earthworm,
Pheretima postuma. Earthworms were divided into eight groups consisting of 4 in each. Group 1 served as
normal control which received saline water only. Group 2 received the standard drug, Albendazole at a dose
level of 10 mg/ml. Groups 3 to 8 received doses of ethanolic extract of 10mg/ml, 20mg/ml, 30mg/ml, 40mg/ml,
50mg/ml and 60mg/ml respectively. Observations were made for time taken to cause paralysis and death of
individual worms in two hours. Paralysis was said to occur when the worms do not revive even in normal saline
water. Death was concluded when the worms lost their motility followed with fading away of their body
colours.
Statistical analysis
The data on biological studies were reported as mean Standard deviation (n = 4). For determining the
statistical significance, standard error mean and analysis of variance (ANOVA) at 5% level significance was
employed. P < 0.05 was considered significant[18].
Table 1: Results of different chemical group tests of the ethanolic extract of dried leaves of Passiflora foetida
Linn.
Ethanloic extract of dried leaves of
Presence
Passiflora foetida Linn.
Reducing sugar -
Steroids -
Alkaloids +
Tannins +
Flavonoids +
Glycosides +
Saponins -
Table 2: Peak list & densitogram of Passiflora foetida Ethanolic extract at UV 366nm with Rf values of
the spots
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50
Series1
40
Series2
30
20
10
0
1 2 3 4 5 6 7 8
Groups
Conclusion
The phytochemical screening on quantitative analysis shows that the leaves of the Passiflora foetida Linn. are
rich in popular phytochemical constituents such as alkaloids, glycosides ( cyanogenetic glycosides), flavonoids
and tannins. The present study has confirmed that the ethanolic extract of leaves of Passiflora foetida Linn.
possess better anthelmintic activity when compared with the standard drug, albendazole. Further studies are
required to isolate and reveal the active compound present in the crude extract of Passiflora foetida L. and to
establish the mechanism of action responsible for anthelmintic activity. An account of the finger print HPTLC
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of ethanolic extract of leaves will definitely help in future for quality check and ensure the genuineness of drug.
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Acknowledgement
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We the authors are thankful to Dr. J. Venkateswara Rao, Principal Sultan- Ul -Uloom College of Pharmacy for
providing necessary facilities and encouragement during the research work.
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