Investigations on Calliandra haematocephala flowers extract for in-vitro anthelmintic

Abstract

Objective: To evaluate the preliminary phytochemical analysis and in-vitro anthelmintic activity of �owers extract of
Calliandra haematocephala. C. haematocephala is an annual, stiff-erect herb found commonly as a weed throughout
India. Different parts of plant is reported to possess phenolic compounds, �avonoids, carbohydrates, alkaloids,
glycosides, saponins, steroids and tannin as major phytochemical groups. Chemical ingredients including phydroxybenzoic
acid, protocatechuic acid, caffeic acid, astilbin, neo-isoastilbin and a fraction yielded lupeol and
betulinic acid. Plant genus shows various pharmacological properties: Anti-in�ammatory, immunomodulatory,
anticonvulsant and antiulcerogenic activity.Materials andmethods: Preliminary phytochemical analysiswas done by
adopting the standard methods. Anthelmintic assay is performed on earthworm (Eisenia foetida) tested for different
concentration (20, 40, 60 and 80 mg/ml) of �ower extract. The effect was compared with standard drug Albendazole.
Result: The present study reveals the presence of few secondary metabolites in the hydroalcoholic extract exhibits
potent antihelmenthic activity.The effectwas observed in dose dependentmanner.Conclusion: Fromthe present study
it can be concluded that hydroalcoholic extract of Calliandra haematocephala �ower possessed signi�cant (P<0.050)
in-vitro anthelmintic effect.
Keywords: Calliandra haematocephala, anti-helmenthic, anti-in�ammatory, laxative

Full text

Introduction
Herbal drugs, referred to as Botanical Medicine, are uses from
plants for their important or medicinal value. An herb is a plant
or plant part valued for its medicinal, aromatic or savory
qualities. Herbals contain a variety of chemical substances that
act upon the body (Chatterjee and Pakrashi, 1997).
Anthelmintics are drug that used to destroy parasite worms
(known as helminthes) from the human body. It has been
expected that about half of the world's population suffers from
helminthiasis and the quantity can be increases. It is not only
poor to tropical and subtropical countries, but is also endemic in
several regions because of poor hygiene, poor family hugiene
malnutrition and crowed living condition (Kiritikar and Basu,
1999).
Calliandra haematocephala, (family: Mimosaceae)
due to its distinct appearance as that of powder puff, it is
commonly known as red powder puff. It is a rambling shrub
with height of 1.5-4 meters has branched pinnates and silky
leaves. The red powder puff flowers are attractive to
butterflies and hummingbirds but only appear during
November-April (Kiritikar and Basu, 1999). The presence
.
of antibacterial and insecticidal properties (Parekh and
Chanda, 2007) in leaves and seeds from former researches
has drawn attention in investigating anthelmintic activities
in flowers as no earlier researches are reported on its flowers
part. is a branched herbal plant, heightC. haematocephala
from 1 to 3 meters. Its branches are brown, cylindric and
rough. Leaves are oblong, acute, with 5 to 10 pairs of glossy
green leaflets. It can be grown in gardens and parks and
found native to tropical America and India. Phytochemical
investigations of the have beenC. haematocephala
extensively done in last few decades. Different parts of plant
are reported to presence of flavonoids, carbohydrates,
alkaloids, glycosides, saponins, steroids and tannin as major
category of compounds. This plant is traditionally used as
Investigations on flowers extract for anthelminticCalliandra haematocephala in-vitro
activity
Jharna Tiwari, Ajay Shukla
Department of Pharmacy, Guru Ram Das Khalsa Institute of Science & Technology, Jabalpur (M.P.) India 483001
* Corresponding Author:Address for
Ajay Shukla
Department of Pharmacy,
Guru Ram Das Khalsa Institute of Science & Technology, Jabalpur,
India
Email: ashukla1007@gmail.com
Abstract
Objective: To evaluate the preliminary phytochemical analysis and anthelmintic activity of owers extract ofin-vitro
Calliandra haematocephala. C. haematocephala is an annual, stiff-erect herb found commonly as a weed throughout
India. Different parts of plant is reported to possess phenolic compounds, avonoids, carbohydrates, alkaloids,
glycosides, saponins, steroids and tannin as major phytochemical groups. Chemical ingredients including p-
hydroxybenzoic acid, protocatechuic acid, caffeic acid, astilbin, neo-isoastilbin and a fraction yielded lupeol and
betulinic acid. Plant genus shows various pharmacological properties: Anti-inammatory, immunomodulatory,
anticonvulsant and antiulcerogenic activity. : Preliminary phytochemical analysis was done byMaterials and methods
adopting the standard methods. Anthelmintic assay is performed on earthworm ( ) tested for differentEisenia foetida
concentration (20, 40, 60 and 80 mg/ml) of ower extract. The effect was compared with standard drug Albendazole.
Result: The present study reveals the presence of few secondary metabolites in the hydroalcoholic extract exhibits
potent antihelmenthic activity.The effect was observed in dose dependent manner. : From the present studyConclusion
it can be concluded that hydroalcoholic extract of ower possessed signicant ( <0.050)Calliandra haematocephala P
in-vitro anthelmintic effect.
Keywords: Calliandra haematocephala, anti-helmenthic, anti-inammatory, laxative
Research Article
17
Received: 14 February 2016 Revised: 10 March 2016 Accepted: 30 March 2016
Advance Pharmaceutical Journal 2016; 1(1): 17-20
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anti-oxidant and as blood purifier (Moharran, 2006). Ethyl
acetate fraction of barks consist of p-C. haematocephala
hydroxybenzoic acid, protocatechuic acid, caffeic acid, astilbin,
neo-isoastilbin lupeol and betulinic acid. The plant genus
showed various pharmacological properties e.g. anticonvulsant,
anti-inflammatory, immunomodulatory and antiulcerogenic
activity. It's butanolic extract reported for gastroprotective
effects in acute gastric lesions (A de Paula et al., 2012). It also
used traditionally as antibacterial (Nia et al., 1999). Recently, C.
haematocephala effervescent granules have been formulated for
the treatment of gastric ulcer treatment (Gupta et al., 2013). In
present study, hydroalcoholic and aqueous extracts of C.
haematocephala flowers were screened for anthelmitic effect on
earthworm.
Materials and methods
Collection, identification and authentication of plant
materials
The fresh flowers of were collected from theC. haematocephala
native area of Jabalpur (M.P.). Plant materials were identified by
the taxonomist from Jawaharlal Nehru Krishi Vishwavidyalaya,
Jabalpur (M.P.). Plant material was dried in shed and powdered
for extraction with alcohol.
Extraction phytochemical analysis of flowers
The flowers of were firstly made free ofC. haematocephala
insects. It was observed that the flowers were very attracting for
insects, variety of insects was found to be stuck on the flowers.
Flowers were shade dried, powdered and were subjected to
extraction with soxhlet apparatus using hydro alcohol as solvent.
Water is another solvent was selected for extraction by
maceration method. The extracts were filtered through the
simple filter paper and concentrate in vacuum under reduced
pressure. The extract were stored in coolC. haematocephala
place for further uses (Dev et al., 2015; Bajaj et al., 2012).
Various qualitative chemical tests were applied in
hydroalcohalic and aqueous extract of Calliandra
haematocephala flowers for detection and confirmation of
different chemical constituents (Kokate et al., 2001; Khan et al.,
2014).
Antimicrobial activity
Antimicrobial activity was evaluated by employing 24 hrs of
culture of using nutrient broth. TheStaphylococcous aureous
microbial strains were transferred to sterile plates aseptically.
The pates were left at room temperature and allowed for
solidification. In each plate one well with 6 mm diameter were
made using a sterile borer. Accurately, 100 μL of different
dilutions of hydroalcoholic extract of andC. haematocephala
single concentration of Ciprofloxacin (10μ/mL) solution, were
transferred to wells aseptically and labeled accordingly. The
plates wereincubated at 37±1 Cfor 24 hours. Thediameters
o
of zone of inhibition were recorded.
Anthelmintic activity
Helminths are worm-like organisms living in and feeding
on living hosts. They receiving nourishment and protection,
while disrupting their host's nutrient and causing weakness.
Those that live inside the digestive tract are called intestinal
parasites. They can live inside human and other animals.
They often find their way into host through contaminated
food or water, soil, mosquito bites and even sexual acts.
Poorly washed vegetables eaten raw may contain eggs of
nematodes such as etc (Gupta et al.,Ascaris, entrobius
2015).
The earthworms were collected fromEisenia foetida
aroud the college campus and authenticated from
Department of Agronomy, Jawaharlal Nehru Krishi
Vishwavidyalaya, Jabalpur (M.P.), India. Four groups of
approximately equal size earthworms consisting of six
earthworms in each group were used for the present study.
Albendazole (20, 40, 60, 80 mg/mL approx) was used as
reference standard. Albendazole, aqueous and
hydroalcohalic extracts were selected in dose of 20 mg/ml,
40 mg/ml, 60 mg/ml and 80 mg/ml concentration.
Observations of the each group were made for the time
taken to paralysis and death of individual worms. Paralysis
was said to occur when the worms do not revive even in
normal saline. Death was concluded when the worms lost
their motility followed with fading away of their body color
(Gupta et al., 2013).
Results and discussion
Preliminary phytochemical analysis
The yield of hydroalcoholic and aqueous extract were
found as 6.37% and 4.51% w/w respectively. Preliminary
phytochemical analysis of hydroalcolic extract exposed the
presence of alkaloids, tannins, flavonoids and glycosides.
Aqueous extract was confirmed the presence of amino acid,
sugars and flavonoids.
Antimicrobial activity
Antimicrobial activity was tested in hydroalcoholic and
aqueous extract of against gramC. haematocephala
positive bacteria . TheStaphylococcous aureous
hydroalcoholic extract of inhibitedC. haematocephala
maximum the growth of bacteria at 100μg/ml and the zone
of inhibition was found 12 mm (Figure 1 and 2). Hence the
hydroalcoholic extract of was mostC. haematocephala
potent against at theStaphylococcous aureous
concentration 100μg/ml (Table 1).
Anthelmintic activity
18
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Advance Pharmaceutical Journal 2016; 1(1): 17-20

The observations of the study (Table 2) was confirms the potent
anthelmintic effect of hydroalcoholic extract of C.
haematocephala. Aqueous extract was observed as lower
anthelmintic activity against tested worms. Observations showed
variable death time and paralysis time at different concentrations.
It is close to the standard drug Albendazole (20, 40, 60, 80,
mg/ml) activity. The activities of the crude extract increase with
increasing the amount or concentration of flowers extract. The
hydroalcohalic extract of flowers having less time in paralysis
and death time of earthworms compared to aqueous extract of
flowers (Figure 3). So that, the hydroalcohalic extract of C.
haematocephala shows significant anthelmintic activity greater
than aqueous extract at maximum concentration.
Table 1. Antimicrobial activity of Calliandra haematocephala
extracts
Value represents as mean ±S.D, n = 3
Table 2. Anthelmintic activity of different extracts of Calliandra
haematocephala
Value represents as mean ±S.D, n = 3
Discussion and conclusion
The potential for developing antimicrobials and anthelmintic
agents from plants appears rewarding in development of
phytomedicine. Results of the investigations cleared that flower
of likely to have phytoconstituents suchC. haematocephala
as alkaloids, tannins, glycosides and flavonoids. These
categories of compound may have properties like
antimicrobial and anthelmintic. This study revealed
antibacterial activity against , testStaphylococcous aureous
micro organism which is etiological agents in various
pathogenic diseases and could be useful in number of
infectious diseases. could serve asC. haematocephala
selective agent for bacterial diseases, chemotherapy and
control. This investigation has opened up the possibility of
use of plant in drug development for human consumption
possibly for treatment of various infectious diseases.
Figure 1. Zone of inhibition observed from test samples of
Calliandra haematocephala extracts
Figure 2. Antimicrobial activity of hydroalcoholic extract of
Calliandra haematocephala
Figure 3. Anthelmintic activity of different extracts of Calliandra
haematocephala
19
Test groups Concentration
(µg/mL)
Zone of inhibition (mm)
Hydroalcoholic
extract
100 12.21 ±0.05
50 10.26±0.06
25 8.61±0.04
Aqueous
extract
100 9.30±0.02
50 8.29±0.03
25 5.40±0.05
Treatment groups Concentration
(mg/ml)
Paralysis
time (min)
Death time
(min)
Hydroalcohalic extract
of C. haematocephala
20 48.60±0.26 64.2±0.341
40 32.32±1.08 58.0± 0.76
60 28.30±0.68 40.13±0.87
80 22.18±0.26 38.46±0.21
Aqueous extract of C.
haematocephala
20 68.60±0.26 94.2±0.40
40 52.32±1.08 78.0±0.86
60 48.30±0.68 70.13±0.76
80 42.18±0.26 88.46±0.23
Albendazole 20 11.00±4.50 22.00±1.8
40 11.60±4.75 45.00±1.00
60 14.10±5.77 47.16±8.23
80 14.30±5.86 49.16±4.00
0
2
4
6
8
10
12
14
0 20 40 60 80 100 120
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e
o
f
i
n
h
i
b
i
i
o
n
(
m
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)
conce ntar tion ( µg/mL )
zone of inhibiton
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Advance Pharmaceutical Journal 2016; 1(1): 17-20

Preliminary phytochemical screening of aqueous and
hydroalcohalic extract of showed theC. haematocephala
presence of flavonoid, glycosides and tannins. Chemically
tannins are poly phenolic compound. Some synthetic phenolic
anthelmintic e.g. niclosamide, oxyclozanide and bithionol are
shown to interfere with energy generation in helmints parasites
by uncoupling oxidative phosphoryation. It is possible that
tannins contained in the extract of mayC. haematocephala
produce similar effects. Another possible anthelmintic effect of
tannins is that they can bind to free protein in the gastrointestinal
tract of host animals. From the result, it can be concluded that the
hydroalcoholic extract of showed significantC. haematocephala
anthelmintic activity and comparable with standard drug. The
extract may be further explored for its phytochemical profile
identifies the active constituents responsible for anthemintic
property.
Acknowledgement
Authors are thankful to Principal, Department of Pharmacy,
G.R.K.I.S.T. Jabalpur, M.P., India, for providing excellent
research facilities and for their cooperation and encouragement
in carrying out this work.
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