K.Hemamalini et al. Int. Res. J. Pharm.

2013, 4 (8)
INTERNATIONAL RESEARCH JOURNAL OF PHARMACY
www.irjponline.com ISSN 2230 – 8407
Research Article

ANTHELMINTIC ACTIVITY OF METHANOLIC LEAF EXTRACT OF SOPHORA INTERRUPTA

K.Hemamalini1*, A.Rajani2, Uma Vasireddy3 and E. Ratna sundari4
1
HOD, Department of Pharmacology, Teegala Ram Reddy College of Pharmacy, Meerpet, India
2
Assistant Professor, SreeDattha Institute of Pharmacy, Sheriguda, Ibrahimpatnam, Greater Hyderabad, India
3
IPA Treasurer, Principal KPS College of Pharmacy, India
4
Dept. of Pharmaceutics, Dhanavanthri College of Pharmacy, Kothagudem, AP, India
*Corresponding Author Email: [email protected]

Article Received on: 15/06/13 Revised on: 07/07/13 Approved for publication: 01/08/13

DOI: 10.7897/2230-8407.04828
IRJP is an official publication of Moksha Publishing House. Website: www.mokshaph.com
© All rights reserved.

ABSTRACT
Crude methanolic extracts of leaves of Sophora interrupta (Fabaceae) were evaluated for in vitro anthelmintic activity on the Indian adult earthworms
Pheritima posthuma. Albendazole was used as a standard drug to compare the test results. The results were interpreted as time taken for paralysis and death of
the earth worms. The study concluded that the methanolic leaf extract of Sophora interrupta showed significant Anthelmintic activity in a dose dependent
inhibition of spontaneous motility of earthworms.
Keywords: Sophora interrupta, anthelmintic, leaf, methanolic, Pheretima posthuma

INTRODUCTION of the institute TRR College of Pharmacy. The leaves of

Gastro intestinal parasites create a serious threat to the Sophora interrupta were dried in the shade, milled into
production of live stock in developing nations1. coarse powder by a mechanical grinder and stored in air tight
Anthelminthics are those agents that expel parasitic worms closed container for further use.
(helminthes) from the body, by either stunning or killing
them. Helminthes parasite infections are global problems Preparation of the Plant Extract
with severe social and economic repercussions in the third The air dried coarse powder of the leaves of Sophora
world countries2. The diseases affect the health status of a interrupta was extracted with methanol using soxhlet’s
large fraction of human population as well as animals. Some apparatus. The powdered material (2 kg) was defatted with
type of dangerous helminthes infections like filariasis has petroleum ether (60-80°C) in a soxhlet extraction apparatus
only a few therapeutic modalities at present3. Helminthes and marc was extracted with methanol (1000 mL) over night,
infections are commonly found in community and being at room temperature with constant stirring. The extract was
recognized as cause of much acute as well as cattle’s use of filtered and the filtrate was concentrated at 30°C under
herbs could be one of the major options to control these reduced pressure in a rotary evaporator. The crude extract
pathologies. The literature survey reveals that Sophora was dissolved in distilled water to required concentrations
interrapta is used to treat various types of gastrointestinal and used for the experiments. The crude extracts were
problems. Therefore an attempt has been made to evaluate subjected to qualitative tests for the identification of various
anthelmintic activity of leaves on adult earthworm Pheritima active constituents. From the dried extract, accurately 5 mg /
posthuma. Sophora interrupta belongs to the family fabaceae ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg /
is commonly called as Edwardsia maderaspatana Wight, Pili ml suspensions of methanolic extract of Sophora interrupta
Girgoli. There are approximately 219 species in genus in normal saline was prepared.
Sophora. Sophora interrupta is available exclusively in
Seshachalam hill ranges of Tirumala, India. This plant is Phytochemical Screening
woody perennial shrub with pinnate leaves, sub opposite leaf- Phytochemical screenings were performed using standard
lets broadly ovate and golden yellow flowers. It has procedures as follows8,9.
multifarious medicinal properties4,5. The plant was
investigated and was found out to possess abortifacient, anti- Test for Reducing Sugars (Fehling’s test)
bacterial, anti-cholesterolemic, anti-inflammatory, anti- The aqueous ethanol extract (0.5 g in 5 ml of water) of
spasmodic, diuretic, emetic, emollient, febrifuge, individual plants was added to boiling Fehling’s solution (A
hypotensive, purgative, styptic and tonic properties6. and B) in a test tube. The solution was observed for a colour
reaction.
MATERIALS AND METHODS
Collection and Authentication of Plant Test for Anthraquinones
The leaves of Sophora interrupta were collected from The individual plant extract (0.5 g) was boiled with 10 ml of
surroundings of Seshachalam hill ranges of Triumala, sulphuric acid (H2SO4) and filtered while hot. The filtrate
Tripathi, Andhra Pradesh, India during the month of August. was shaken with 5 ml of chloroform. The chloroform layer
The plant material was taxonomically identified and was pipette into another test tube and 1 ml of dilute ammonia
authenticated by Dr. Madhava Chetty, Associate professor, was added. The resulting solution was observed for colour
Department of Botany, S.V. University, Tripathi7 and a copy changes.
has been preserved for the future reference at the herbarium

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K.Hemamalini et al. Int. Res. J. Pharm. 2013, 4 (8)
Test for Terpenoids (Salkowski test) Test for Cardiac Glycosides (Keller-Killiani test)
To 0.5 g each of the individual extract was added 2 ml of To 0.5 g of individual plant extract diluted to 5 ml in water
chloroform. Concentrated H2SO4 (3 ml) was carefully added was added 2 ml of glacial acetic acid containing one drop of
to form a layer. A reddish brown coloration was confirmed ferric chloride solution. This was under laid with 1 ml of
for the presence of terpenoids. concentrated H2SO4. A brown ring at the interface indicated
the presence of a deoxysugar characteristic of cardenolides. A
Test for Flavonoids violet ring may appear below the brown ring, while in the
A portion of the individual plant extract (0.5 g) was heated acetic acid layer a greenish ring may form just above the
with 10 ml of ethyl acetate over a steam bath for 3 minutes. brown ring and gradually spread throughout this layer.
The mixture was filtered and 4 ml of the filtrate was shaken
with 1 ml of dilute ammonia solution. A yellow coloration Animals
indicates the presence of flavonoids. Pheretima posthuma (Adult Indian earth worms) of about 5-7
cm long were used for the present study.
Test for Saponins
To 0.5 g of each plant extract was added 5 ml of distilled Standard Drug used
water in a test tube. The solution was shaken vigorously and Albendazole suspension (micronized albendazole suspension
observed for a stable persistent froth. The frothing was mixed in the concentration of 10 mg / ml) was used as the standard
with 3 drops of olive oil and shaken vigorously after which it to compare the test results.
was observed for the formation of an emulsion.
Anthelmintic Activity
Test for Tannins Pheretima posthuma (Indian adult earth worms) of nearly
About 0.5 g of the individual extract was boiledin 10 ml of equal size (6 cms ± 1) were selected randomly for the present
water in a test tube and then filtered. A few drops of 0.1 % study10-12. The worms were acclimatized to the laboratory
ferric chloride (FeCl3) was added and observed for brownish conditions before experimentation. The earth worms were
green or a blue-black coloration. divided into four groups of six earth worms in each.
Albendazole suspension in the concentration of 10 mg / ml
Test for Alkaloids served as a standard and poured into petri dishes. The test
0.5 g of each extract was diluted to 10 ml with acid alcohol, extract were prepared in the concentrations of 5 mg / ml, 10
boiled and filtered. To 5 ml of the filtrate was added 2 ml of mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml.
dilute ammonia. 5 ml of chloroform was added and shaken Normal saline served as control. Six earth worms nearly
gently to extract the alkaloid base. The chloroform layer was equal size 6 cms ± 1 were taken for each concentration and
extracted with 10 ml of acetic acid. This was divided into two placed in petri dishes at room temperature13. The time taken
portions. Mayer’s reagent was added to one portion and for complete paralysis and death were recorded. The mean
Draggendorff’s reagent to the other. The formation of a paralysis time and mean lethal time for each sample was
cream (with Mayer’s reagent) or reddish brown precipitate calculated. The time taken for the worms to be become
(with Draggendorff’s reagent) was regarded as positive for motionless was noted as paralysis time and to ascertain death,
the presence of alkaloids. each worm was frequently applied with external stimuli
which stimulates or induce movements in the earthworm, if
alive14.

Table 1: Anthelmintic Activity of Methanolic Extract of Sophora interrupta leaves

Treatment Concentration used (mg/ml) Timetaken for paralysis (min) X ± S.D Time taken for death (min) X ± S.D
Control --- --- ---
Standard 10 mg / ml 17 ± 1.571* 39 ± 1.932*
(Albendazole)
Methanolic extract 5 mg / ml 76 ± 3.303* 99.33 ± 0.402*
of 10 mg / ml 65.33 ± 2.883** 90.67 ± 3.921**
Sophora interrupta 15 mg / ml 56 ± 2.017** 85.00 ± 5.310**
20 mg / ml 4.33 ± 1.498** 69.83 ± 2.496**
25 mg / ml 48.33 ± 1.498** 51.67 ± 2.108**
30 mg / ml 27.33 ± 2.060** 40.33 ± 0.9888***
All values are Mean ± SEM analyzed by one way ANOVA followed by Dunnett’s test. *p < 0.05,** p < 0.01,*** p < 0.001

RESULTS concentration of 30 mg / ml has taken less time to cause

The earlier studies on preliminary phytochemical paralysis, and little more time to cause death of earthworms
investigations of Sophora interrupta plant revealed the as compared with reference drug.
presence of alkaloids, cardiac glycosides, tannins, saponins,
flavonoids and poly phenolics. Methanolic extracts of DISCUSSION
Sophora interrupta were used to evaluate anthelmintic Some of the traditionally used herbs have scientifically
activity and showed the effect in a dose dependent manner. proved a potent anthelmintic activity by using suitable
The mean ± SEM values (statistical analysis) were calculated experimental models. The predominant effect of Albendazole
for each extracts. The results of anthelmintic activity on earth on the worm is to cause flaccid paralysis that result in
worm Phertima prosthuma was given in Table 1, reveal that, expulsion of the worm by peristalsis. Albendazole by
the different concentration of the extracts has shown paralysis increasing chloride ion conductance of worm muscle
and death of earthworms and it was compared with membrane produces hyperpolarization ad reduced excitability
albendazole as reference drug. Methanolic extract in the that leads to muscle relaxation and flaccid paralysis. The

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K.Hemamalini et al. Int. Res. J. Pharm. 2013, 4 (8)
extract demonstrated paralysis as well as death of the worms REFERENCES
at a time comparable to Albendazole especially at higher 1. Kuppast IJ, Nayak V. Anthelmintic activity of fruits of Cordia
Dichotoma. Indian journal of natural products 2003; 19(3): 27-29.
concentration of 30 mg / ml. Poly phenolic compounds 2. Srinivasa MV, Jayachandran E. Amthelmintic activity of 8-fluoro-9-
shown anthelmintic activity; chemically tannins are poly substituted (1,3)-benzthiazole (5, 1-B)- 1,2,4 triazole on Pheretima
phenolic compounds. Some synthetic phenolic anthelmintics, posthuma. Indian drugs 2006; 43(4): 343-347.
Eg: - Niclosamide, Oxyclozanide and Bithionol are shown to 3. Maity TK, Mandal SC, Mukherjee PK, Saha K, Das J. Studies on Anti-
inflammatory effect of Cassia tora leaf extract (Fam. Leguminoceae).
interface with energy generation in helminth parasites by Phytotherapy research 1998; 12(3): 221-223. http://dx.doi.org/10.1002/
uncoupling oxidative phosphorylation. It is possible that the (SICI)1099-1573(199805)12:3<221::AID-PTR221>3.0.CO;2-L
active principle like tannins in the extract of Sophora 4. Deena Dalith M. Anti-oxidant activity of Sophora interrupta.
interrupta produces similar effects. Another possible International journal of phyto pharmacology 2011; 2(2): 43-47.
5. Tse WP, Che CT, Liu K and Tin ZX. Evaluation of Anti-proliferative
anthelmintic effect of tannins is that they can bind to free properties of selected psoriasis treated Chinese medicines on cultured
protein in the gastro intestinal tract of host animal or glycol HaCa T cells. Journal of Ethno pharmacology 2006; 108: 133-141.
protein on the parasite and cause death15. Further the extract http://dx.doi.org/10.1016/j.jep.2006.04.023 PMid:16730935
can be tested on various other helminthes to ascertain the 6. Poretz RD, Barth RF. Studies on the interaction of the Sophora japonica
lectin and concanavalin A with erythrocytes and lymphocytes. R.
anthelmintic activity on a broader scale which is our future Immunology 1976; 31: 187.
plan of research work. 7. Madhavachetty K, Sivaji K, Tulasi Rao K. Flowering plants of chitter
district. AP; 2008.
CONCLUSION 8. Trease GE, Evans WC. Pharmacognosy. London: 13th ed. Bailliere
Tindall; 1989. p. 176-180.
From the above result it is concluded that methanolic extracts 9. Pandey Govind, Madhuri S. Medicinal plants: better remedy for
of Sophora interrupta have a potent anthelmintic activity neoplasm. Indian Drugs 2006; 43: 869-874.
when compared with conventionally used drug. It is 10. Dash GK, Suresh P, Kar DM, Ganpaty S, Panda SB. Evaluation of
comparable with standard drug. Further studies using in vivo evaluation salsinoids Linn. for Anthelmintic and Anti-microbial
activities. J. Nat Rem 2002; 2: 182-5.
model are required to find out and to establish effectiveness 11. Szewezuk VD, Mongelli ER, Pomilio AB. Anti-parasitic activity of
and pharmacological rationale for the use of leaves as Melia azadirach growing in Argentina. Molecular Med Chem 2003; 1:
anthelmintic drug. Further studies to isolate active constituent 54-5.
from extracts to establish (s) mechanism of action are 12. Shivkar YM, Kumar VL. Anthelmmintic activity of latex of Callotropic
procera. Pharma Biol 2003; 41: 263-265. http://dx.doi.org/
required. 10.1076/phbi.41.4.263.15666
13. Kaushik RK, Katiyar JC, Sen AB. Studies on the mode of the action of
ACKNOWLEDGEMENTS Anthemintics with Ascardiagalli as a test parasite. Indian J Med Res
Authors are thankful to the management of Teegala Ram Reddy College of 1974; 62: 1367-1375. PMid:4442996
Pharmacy for providing us the required facilities to carry out the research 14. Lal J, Chandra S, Raviprakash V, Sabir M. In vitro Anthemintic action
activity. of some indigenous medicinal plants on Ascardiagalli worms. Indian
Physiol Pharmacol 1976; 20: 64-68,
15. Mali RG, Wadekar RR. Indian J Pharm Sci 2008; 70: 131-133. http:/
/dx.doi.org/10.4103/0250-474X.40352PMid:20390101 PMCid:PMC28
52054

Cite this article as:

K.Hemamalini, A.Rajani, Uma Vasireddy and E. Ratna sundari.
Anthelmintic activity of methanolic leaf extract of Sophora interrupta. Int.
Res. J. Pharm. 2013; 4(8):148-150 http://dx.doi.org/10.7897/2230-
8407.04828

Source of support: Nil, Conflict of interest: None Declared

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