ANTHELMINTIC ACTIVITY OF ETHANOL AND ETHYL ACETATE
EXTRACTS OF TECOMARIA CAPENSIS. (FAMILY:
BIGNONIACEAE)
E. Tamil Jothi1, Ch. Harini2* and G. Nagarjuna Reddy3
1
Department of Pharmacology, K.L.R Pharmacy College, Palvoncha, Telangana.
2
Doctor of Pharmacy, K.L.R Pharmacy College, Palvoncha, Telangana.
ABSTRACT
Ethanol and ethyl acetate extract of Tecomeria capensis
(Bignoniaceae) was investigated for the activity against Indian
earthworms Pheretima posthuma. Various concentrations of extract
(25-100mg/ml) were tested, which involved determination of time of
paralysis and time of death of the worms. Ethanol and ethyl acetate
extract at the tested dose were produced significant activity. The
maximum activity of ethanol and ethyl acetate extract of Tecomeria
capensis was 8.55 and 8.50 minutes respectively as time of paralysis,
32.25 and 32.10 minutes respectively as time of death at the dose of
100 mg/ml. Albendazole (20 mg/ml) which is included as standard reference showed 31.86
minutes as time of paralysis, 42.5 minutes as time of death. 0.5% w/v of carboxymethyl
cellulose used as control there is no paralysis and death of earth worms. The present study
indicates ethanol and ethyl acetate extract of Tecomeria capensis acts as potential
anthelmintic agent.
KEYWORDS: Albendazole, Anthelmintic activity, Pheretima posthuma, Paralysis time, Death time, Tecomeria capensis.
1. INTRODUCTION
Parasitic diseases are a major infestation in the human beings like helminthiasis. The disease
is caused by round worm, hook worm, thread worm, tape worm and filarial, guinea worm are
found in intestine.[1] The worm is responsible for many types of diseases; they harm the host by depriving of food, causing blood loss in stool, injury to organs, intestinal or lymphatic
Volume 5, Issue 1, 1443-1449. Research Article ISSN 2277– 7105
Article Received on 18 Nov 2015,
Revised on 09 Dec 2015, Accepted on 29 Dec 2015
*Correspondence for
Author
Dr. Ch. Harini
Doctor of Pharmacy,
K.L.R Pharmacy College,
ill health. Number of synthetic drugs used to control and prevent the infestation related to
worms, the drugs like mebendazole, albendazole, piperazine and pyrantel, almost
mebendazole used as broad spectrum anthelmintic drugs.[2] Adverse effect like tolerance, resistance, nausea, vomiting, drowsiness, dizziness and abdominal pain occurred at long term
used of synthetic medicine. Therefore, overcome the problem associated with synthetic
medicine, the natural compounds are selected. Naturally produced medicinal products offer
as an alternate anthelmintic and therapeutic agents so as to overcome some of the infestation
and subsequently may be sustainable and environmentally acceptable because the natural or
herbal compounds are free from adverse effect.[3]
However, No scientific data are available regarding ethyl acetate and ethanol extracts of
Tecomaria capensis (leaves) usefulness as anthelmintic agent. Keeping the above information
in view the present study was endeavored to ratify the anthelmintic activity of the extracts of
Tecomaria capensis on Indian earth worms (Pheretima posthuma).
2. MATERIALS AND METHODS 2.1. Plant material
The leaves of Tecomaria capensis (Thunb.) Spach were collected from Guntur district, A.P
and it was authenticated by professor Dr. S.M. Khasim, Department of Botany and
Microbiology, Acharya Nagarjuna University, Guntur. The specimen (No:
ANU/00129/2009/AP) was deposited in the department of botany and microbiology for
future reference. Fresh plant material was collected in bulk, washed under running tap water
to remove adhering material, dried under shade and pulverized in a mechanical grinder. The
coarse powder was passed through sieve no. 60#. Care was taken to select healthy plants and
for normal organs.
2.2. Preparation of extracts Ethanol extract
This extract was prepared by using A Soxhlet apparatus. About 150gm of dried flower
powder was taken In a muslin cloth bag. The purified Ethanol was passed through the tube
where the powder bag was kept. The Ethanol was passed through siphon tube so that it
reaches the round bottom flask in which porcelain chips were provided. The vapours
containing the constituents pass through the condenser and reach the tube containing powder
flask containing extract was transferred to a beaker and was allowed to evaporate in a water
bath. This concentrated Ethanol extract was used for further studies.
Ethyl acetate extract
This extract was prepared by using Soxhlet apparatus. About 150gm of dried flower powder
was taken in a muslin cloth bag. The purified Ethyl acetate was passed through the tube
where the powder bag was kept. The Ethyl acetate was passed through siphon tube until it
reached the round bottom flask in which porcelain chips were provided. The vapours
containing the constituents passed through the condenser and reached the tube containing
powder bag and the process was repeated. This was continued for 24hours. Then the round
bottom flask containing extract was transferred to a beaker and was allowed to evaporate in a
water bath. This concentrated Ethyl acetate extract was used for further studies.
2.3. Drugs and Chemicals used
Albendazole (Glasko Smith Kline) was used as reference Standard purchased from local
medical shop, Thiruninravur, Chennai.
2.4. Administration of Albendazole
Albendazole (20mg/ml) was prepared by using 0.5% W/v of CMC as a suspending agent and
administered as per method of extract.
2.5. Experimental animals
Indian adult earthworms (Pheretima posthuma) were collected from moist soil and water
logged areas at Vadlamudi, Tenali Road and were identified at the Department of Zoology,
JMJ college for women, Tenali. Then all collected worms were washed with normal saline to
remove all the faecal matter and used for the anthelmintic study. The earthworms of 3-5 cm
in length and 0.1 - 0.2 cm in width were used for all the experimental protocol.
2.6. Preparation of test sample
Samples for experiments were prepared by dissolving extract to obtain a stock solution of 200
mg/ml, from the stock solution, different working dilutions were prepared to get
concentration range of 25, 50, 75 and 100 mg/ml. For present study Albendazole has taken as
standard drug. The concentration of standard drug was prepared by using 0.5% w/v of CMC
2.7. Experimental organism Groups Dividing
The Indian adult Earth worms can be divided into ten groups. Each group consist six earth
worms. Group-I is contain Vehicle (0.5% w/v of CMC), Group-II containing Albendazole as
a reference standard (20 mg/ml), remaining groups having ethanol and ethyl acetate extracts
with different concentrations such as 25 mg/ml of ethyl acetate extract as Group III, 50
mg/ml of ethyl acetate extract as Group IV, 75 mg/ml of ethyl acetate extract as Group V,
100 mg/ml of ethyl acetate extract as Group VI, 25 mg/ml of ethanol extract as Group VII, 50
mg/ml of ethanol extract as Group VIII, 75 mg/ml of ethanol extract as Group IX, 100 mg/ml
of ethanol extract as Group X.
2.8. Evaluation of Anthelmintic Activity
The anthelmintic activity was performed according to the method of Ghosh et al.[4] on adult Indian earthworm, Pheritima posthuma as it has anatomical and physiological resemblance
with the intestinal roundworm parasites of human beings.[5,6] Pheritima posthuma worms are easily available and used as suitable model for screening anthelmintic drugs.[7] In the 50 ml of four different concentration of ethyl acetate extract and ethanol extract (25, 50, 75 and
100mg/ml in normal saline) and the standards Piperazine citrate (10 mg/ml) and Albendazole
(20 mg/ml) were prepared and approximately equal sized six earthworms were released in
each group. Observations were made for the time taken to paralyse or 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 lose their motility followed with fading away of their
body color. Piperazine citrate (10mg/ml) and Albendazole (20mg/ml) were used as reference
standards and normal saline water as control.
3. RESULTS AND DISCUSSION
From the observations made, ethyl acetate and ethanol extracts exhibited anthelmintic activity
in dose-dependent manner giving shortest time of paralysis (P) and death (D) with 100 mg/ml
concentration. Results are shown in table no.1. The ethyl acetate extract of T. capensis caused paralysis (P) in 8.50 min and time of death(D) is 32.1 min. while ethanol extract
revealed paralysis in 8.55 min and time of death is 32.25 min respectively against the
earthworm Pheretima posthuma. The reference drug Piperazine citrate showed paralysis in
22.77 min and time of death is 61.01 minutes and Albendazole showed paralysis in 31.86 min
Albendazol by increasing chloride ion conductance of worms muscle membrane produced
hyperpolarization and reduced excitability that which led to muscle relaxation and flaccid
paralysis.[8] Phytochemical screening of the extracts revealed the presence of alkaloids, saponins, flavonoids, triterpenes, tannins and steroids. Tannins were shown to produce
anthelmintic activities chemically tannins are polyphenolic compounds. It is possible that
tannins contained in the extracts of T.capensis produced similar effects. Reported
anthelmintic effect of tannins is that they can bind to free proteins in the gastrointestinal tract
of host animal or glycoprotein on the cuticle of the parasite and may cause death. The
Piperazine citrate acts by increasing chloride ion conductance of worm muscle membrane
produces hyperpolarisation and reduced excitability that leads to muscle relaxation and
flaccid paralysis.
Mechanism of the anthelminthic activity of T.capensis cannot be explained on the basis of
our present results. From the observations made, higher concentration of extract produced
paralytic effect much earlier and the time to death was shorter for all worms. Further studies
are in process to identify the possible phytoconstituents responsible for anthelmintic activity.
Table - 1: Anthelmithic activity of ethanol and ethyl acetate extracts of Tecomaria capensis on Indian Earthworms (Pheretima posthuma).
Test substance
Concentration (mg/ml)
Time taken for Paralysis(min.)
Time takenfor Death(min.)
Vehicle --- No paralysis
(upto72min)
No death (upto 72min) Piperazine Citrate(Std.) 10 22.77±0.27** 61.01±0.42**
Albendazole (Std.) 20 31.86±0.22** 42.50±0.26**
Ethyl acetate extract
25 36.35±0.23** 72.10±0.33** 50 23.59±0.29** 64.1±0.24** 75 18.09±0.07** 43.91±0.26** 100 8.50±0.37** 32.10±0.53**
Ethanol extract
25 34.94±0.93** 69.05±0.28** 50 28.43±0.18* 48.83±0.29** 75 19.29±0.20** 51.6±0.21** 100 8.55±0.10** 32.25±0.25**
Each value represents mean ± SEM (N=6). *P<0.05, **P<0.01. This activity was
Concentration dependent. The potency was found to be inversely proportional to the time
4. CONCLUSION
From the above results, it is concluded that ethyl acetate and ethanol extracts of Tecomaria
capensis, showed significant anthelmintic activity. The experimental evidence obtained in the
laboratory model could provide a rationale for the traditional use of this plant extracts as
anthelmintic activity.
5. ACKNOWLEDGEMENT
The author is thankful to management, teaching and nonteaching staff of K.L.R Pharmacy
College for the facilities generously they are provided to execute some of the research work
presented in the article.
6. CONFLICT OF INTEREST Conflict of Interest declared none.
7. REFERENCES
1. Agbolade O M, Akinboye D O, Awolajam A, Intestinal helminthiasis and urinary
schistosomiasis in some villages of Ijebu North, Ogun State, Nigeria. African J. Biotech,
2004; 3: 206-209.
2. Afroz Jahan, Sanweer Khatoon C R and Iswati A Umap, In Vitro Anthelmintic Activity
of Phyllanthus Niruri Linn. Against Paramphistomes. Biochem. Pharmacol, 2009; 5(3):
836-838.
3. Deore S L, Khadabadi S S, In vitro Anthelmintic activity of Cassia tora. Int. J. of Chem.
Tech. Res, 2009; 1(2): 177-179.
4. Girme AS, Bhalke RD, Ghogare PB, Tambe VD, Jadhav RS, Nirmal SA. Comparative In
vitro Anthelmintic Activity of Mentha piperita a Lantana camara from Western India.
Dhaka Univ Pharm Sci., 2006; 5: 5-7.
5. Tambe VD, Nirmal SA, Jadhav RS, Ghogare PB, Bhalke RD, Girme AS, Bhamber RS.
Anthelmintic activity of Wedelia trilobata leaves. Ind J Nat Prod, 22: 27-29.
6. Dash GK, Sursh P, Sahu SK, Kar DM, Ganpaty S, Panda SB. Evaluation Evolvulus
alsinoids Linn. for anthelmintic and antimicrobial activities. J Nat Rem., 2002; 2: 182-85.
7. Daginawala HF, Prasad S, Kashyap RS, Deopujari JY, Purohit HJ and Taori GM.
Development of an in vitro model to study clot lysis activity of thrombolytic drugs.
8. Dickerson EH, Cho LW, Maguiness SD, Killick SL, Robinson J, Atkin SL. Insulin
resistance and free androgen index correlate with the outcome of controlled ovarian