International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701

_________________________________________Research Article

Anthelmintic activity of Methanolic and Ethanolic Leaf Extract

of Calendula officinalis
Dhawal Dorwal

Department of Pharmaceutics, Rajendra Institute of Technology and Sciences, Sirsa,

Haryana, Chandigarh, India.
__________________________________________________________________________________
ABSTRACT
The methanolic and ethanolic extract of leaves of Calendula officinalis was prepared in three different
concentrations viz 5 mg/ml, 10 mg/ml and 15 mg/ml for investigation of anthelmintic activity in-vitro.
The results suggested that both the extracts showed significant anthelmintic activity as compared to
the standard drug, Albendazole (10 mg/ml) and it was also noticed that higher concentrations depicted
better anthelmintic activity in-vitro.

Key Words: Calendula officinalis, methanolic, ethanolic, Anthelmintic activity.

INTRODUCTION
Calendula officinalis Linn (Asteraceae),
commonly known as Pot Marigold is an
important Medicinal plant used in our
Traditional System of Medicine and has been
widely used in homeopathic medicine for the
treatment of many diseases1. It has been
reported to possess many pharmacological
activities, which include antioxidant, anti-
inflammatory, antibacterial, antifungal and
antiviral. It also possess cytotoxic as well as
tumor reducing potential2. C. officinalis contains
following potentially active chemical
constituents: Sesquiterpene and flavonol
glycosides, Triterpenoid, saponins, Triterpene
alcohols, Flavonoids, carotenoides, xanthophylls,
Phenolic acids and Other like sterols, mucilage,
tocopherols, calendulin, bitters, phytosterols,
resin, volatile oils3. The aqueous extract of
calendula leaves have already been investigated
for their anthelmintic activity in-vitro4.
Therefore, present study focuses on methanolic
and ethanolic extracts of the drug leaves for
investigation of anthelmintic activity in-vitro.
MATERIALS AND METHOD
Drugs and Chemicals
Albendazole ( gift sample from Sigma Aldrich),
DMF, Methanol AR and Ethanol AR were
used during experimental work.
Plant Material
The leaves of Calendula officinalis were
collected from local farm land of Gurgaon,
Haryana. The leaves were washed under
running tap water and dried in hot air oven at
40 degree for 8 days and then homogenized to
get a coarse powder.
Preparation of solvent extract
Powdered leaf (500 g) was successively
extracted with methanol and ethanol using a
Soxhlet apparatus. The extraction was carried
out for 24 hrs at room temperature with mild
shaking. The extracts were filtered and
concentrated at 35° C by rota evaporator. The
extractive values of methanol and ethanol was
found to be 16.5% and 17.1 % respectively1.
Preparation of test and standard sample
Samples for in-vitro study were prepared by
dissolving the all extracts in minimum quantity
of DMF and normal saline so as to get final
concentration of 5, 10 and 15 mg/ml strength
and standard drug, Albendazole of 10 mg/ml.
Animals
Indian adult earthworms (Pheretima posthuma
(Annelida)) 9-11 cms in length and 0.3-0.5 cms
in width were taken for all the experimental
protocol. Indian adult earthworms were obtained
from water logged area of moist soil and
washed with normal saline to remove all faecal
matter.
Anthelmintic Activity
The assay was performed on Indian adult earth
worm5, Pheretima posthuma due to its
anatomical and physiological resemblance with
the intestinal round worm parasite of human
being6. Eight groups of approximately equal
size Indian earthworms consisting of six worms

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 International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701

in each group were released into 50 ml of anthelmintic activity in calendula leaves as

desired formulation as following groups:
Group as control: DMF with normal saline.
Group (Standard): Albendazole at the dose of
10 mg/ml as the standard.
Group M-5, M-10 and M-15: Methanolic
extract at the dose of 5, 10, 15 mg/ml
respectively.
Group E-5, E-10 and E-15: Ethanolic extract at
the dose of 5, 10, 15 mg/ml respectively.
Observations were made from the time taken
for paralysis and / or death of individual
earthworms. The time of paralysis was noted
when worms do not revive even in the normal
saline and when no movement of any sort
could be observed except when the worms were
shaken vigorously. Death was concluded when
the worms lost their motility.
Statistical Analysis
All the results were expressed as a mean ±
SEM of six animals in each group. The data
were subjected to statistical analysis one- way
ANOVA followed by Dunnett post-test was
employed to find significant different means by
using Graph- Pad In Stat computer package.
RESULTS AND DISCUSSIONS
Saponins were confirmed by the phyto-chemical
investigation as they are majorly responsible for
Umesh K Jain et al had found in the aqueous
extract of calendula leaves and flowers and
suggested their anthelmintic activity in-vitro.
The aim of this study was to investigate if
alcoholic extracts of calendula leaves were also
as potent as previously investigated aqueous
extracts of calendula by Umesh K Jain for
anthelmintic activity.
The methanolic and ethanolic extracts of
calendula leaves showed significant anthelmintic
activity. Both the extracts showed dose
dependent decrease in paralysis and death time
as evident from ( table 1 and figure 1) M-5 to
M-15 and E-5 and E-15, as dose was increased
the paralysis and death time decreased. Further,
it may be inferred from data (fig.1) standard
drug, albendazole (10 mg/ml) showed paralysis
time of 29 min. and death time of 44 minutes
comparable with results with E-15 sample as
paralysis time of 35 min. and death time of 46
min.
CONCLUSION
The present research shows promising results
with solvent extraction method for anthelmintic
activity for calendula officinalis. Further, in-
vivo models should be developed for harbouring
the potential of this herbal drug as future
medicine for human kind.

Table 1: Screening of In vitro Anthelmintic Activity of Calendula officinalis.

No. of Paralysis Time Death Time
Groups worms Dose (mg/ml) (min) (min)
used (Mean ± SEM) (Mean ± SEM)
Control 6 - 0 0
Standard 6 10 29±1.73 44±2.02
M-5 6 5 155±2.03 305±1.99
Methanolic
M-10 6 10 95±1.20 185±1.34
extract
M-15 6 15 69±2.20 139±1.29
E-5 6 5 111±1.97 190±1.98
Ethanolic
E-10 6 10 62±0.92 135±1.54
extract
E-15 6 15 35±1.23 46±1.18

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 International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701

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