It will helpful for doing projects in pharmacology

1. EVALUATION OF IN-VITRO ANTI HELMINTHIC POTENCY
OF ETHANOLIC EXTRACT OF TEPHROSIA PURPUREA.
Under the Guidance of
Mrs.K.Lakshmi ,M.Pharm., Assistant Professor, Department of Pharmacology ,
Sri Siddhartha Pharmacy College,
NUZVID-521202,
AUGUST-2021.
mm
Dissertation submitted to
KRISHNA UNIVERSITY, Machilipatnam.
In partial fulfillment for the award of degree of
BACHELOR OF PHARMACY
Damuluri Haritha(Y17BPH203015)
Gokavarapu Anitha(Y17BPH203019)
Vaka Krishna Prasad(Y17BPH203033)
Amudala Ramya(Y17BPH203001)
Bokinala Anusha(Y17BPH203006)
Bondili Deekshitha Singh(Y17BPH203008)
Duvvari Vijay Teja (Y17BPH203016)
7/21/2022 1

2. CONTENTS
 Abstract
 Introduction
 Plant profile
 Plan of work
 Aim and objectives
 Literature review
 Materials and methods
 Results and discussion
 Summary and Conclusion
 References
7/21/2022 2

3. ABSTRACT
Various species of genus Tephrosia have been extensively studied for their
pharmacological activity. Tephrosia purpurea is expected to possess many
constituents among them triterpenoids are major constituents. So due to presence of
triterpenoids Tephrosia purpurea may possess Antihelmintic activity. In the present
work ethanolic extract of leaves of Tephrosia purpurea was evaluated for its
Antihelmintic activity against earthworm at three different concentrations (25, 50,
100 mg/ml). The Antihelmintic activity of extract was comparable with Marketed
allopathic drugs (Piperazine citrate (10 mg/ml), Albendazole (25 mg/ml, 50 mg/ml),
Ivermectin (5 mg/ml), Levamisole (10 mg/ml). 2% v/v Tween 80 in water was
taken as a normal control. Marketed allopathic drugs concentrations were prepared
using 2% v/v Tween 80. Antihelmintic activity was evaluated by considering the
time required for mobility, paralysis and death of worms by all the prepared extracts
and marketed allopathic drugs concentrations.
Levamisole>Piperazine citrate>Tephrosia purpurea leaf extract
>Ivermectin >Albendazole.
Key Words: Tephrosia purpurea, Antihelmintic activity, Earthworms.
7/21/2022 3

4. INTRODUCTION
Helminthiasis (alternatively spelled Helminthiasis;
plural helminthiases) is any macro parasitic disease of
humans and animals in which a part of the body is
infected with parasitic worms known as helminthes.
These parasites are broadly classified
into tapeworms, flukes, and roundworms.
They often live in the gastrointestinal tract of
their hosts, but may also burrow into other organs,
where they induce physiological damage. They remain
the major cause of wildlife diseases, economic crises in
the livestock industry, and human socio-economic
problems in developing countries.
7/21/2022 4

5. PLANT PROFILE
TAXONOMICAL CLASSIFICATION
7/21/2022 5
Kingdom Plantae
Subkingdom Tracheobionta
Division Magnoliophyta
Class Magnoliopsida
Subclass Rosidae
Order Fabales
Family Fabaceae
Genus Tephrosia
Species Purpurea
Part used Leaves
FIGURE 1 -Tephrosia purpurea plant and
leaves

6. PLAN OF WORK
Collection of Tephrosia purpurea leaves
Powdered
Subjected to extraction with non-polar (ethanol) solvent by using
Soxhlation
Drying
Preliminary phytochemical analysis
Comparative Antihelmintic activity against Indian earthworm.
Results and discussions

7. AIM & OBJECTIVES
AIM:
To evaluate the comparative in vitro Antihelmintic activity of Tephrosia purpurea leaves
along with various marketed branded drugs against Indian earthworms.
OBJECTIVES: The objective of the present work is
 To carry out the Leaves of Tephrosia purpurea. extraction with non-polar (Ethanol)
solvent by using Soxhlet extraction.
 To carry out the preliminary phytochemical analysis of ethanolic extract of Tephrosia
purpurea.
 To screen the comparative in vitro Anti helminthic activity of Ethanolic extract of
Tephrosia purpurea leaves in various strengths with marketed brands of Anti helminthic
drugs.
7/21/2022 7

8. REVIEW OF LITERATURE
 Rajender Vadluri et al. (2016) Performed in vitro propagation and production of
Tephrosia purpurea plant via internodal explants. Tephrosia purpurea (Linn.) Pers.
(Leguminasae) is a highly sub erect, branched and herbaceous perennial herb.
 Pankti P Dalwadi et al. (2014) Performed A Review on Phytochemistry and
Pharmacological Studies of Tephrosia purpurea Linn (Sharpunkha, Wild Indigo).
 Suriyavathana Muthukrishnan et al. (2016) Performed in vitro Antioxidant
Potential of Methanolic Extract of Tephrosia purpurea L.
 Anuradha Laishram et al. (2013) Performed Phytochemical analysis, TLC
profiling and antimicrobial activity of Tephrosia purpurea.
 Santram Lodhi et al. (2016) Performed effects of flavonoids from Martynia annua
and Tephrosia purpurea on cutaneous wound healing. Martynia annua L.
 S. Sujatha et al. (2014) Performed integrated imminent wide scientific potential
from tropical weedy medicinal plant of Tephrosia purpurea (linn.) pers.
 S. Sandhya et al. (2010) Performed Tephrosia maxima and Tephrosia purpurea
Pers. is a pan tropical coastal shrub which contain flavonoids and isoflavanones.
 Saad Touqeer et al. (2013) Performed A Review on the Phytochemistry and
Pharmacology of Genus Tephrosia.
7/21/2022 8

9. MATERIALS AND METHODS
Collection & Identification of Plant :-
• Leaves of Tephrosia purpurea were purchased from local market in Hyderabad. A
voucher specimen was kept in department for future reference.
Preparation of extract:-
The plant leaves was initially washed with water and then shade dried in
sunlight for 3 days until leaves became completely dry. The dried leaves were
made into a fine powder by using a mixer grinder. The fine powder was taken
for further extraction procedure. Then weighed amount of 67 gm of powder of
leaves of Tephrosia purpurea was packed in thimble flask and 550ml of ethanol
(70%) was added in 1 litre round bottom flask. Then the Soxhlet assembly was
set up to complete 10 to 15 cycles. After that the extract was filtered and filtrate
was concentrated up to 50 ml using water bath. Filtration of mixture was
carried out. Filtrate was taken in a beaker and evaporated until it converted into
semisolid concentrated liquor. Then it was dried completely to get dried extract.
The % yield was found to be 19.40% w/w.
7/21/2022 9

10. Preliminary phytochemical screening: The preliminary phytochemical
investigation was carried out with ethanolic extract of Tephrosia
purpurea (Linn) for identification of phytochemical constituents.
Phytochemical tests were carried out by standard methods.
Test Organism :
Indian adult earthworms (Pheretima posthuma) were used during
the experiment. The earthworms were collected from a local
supplier. Worms were washed with normal saline to remove all
fecal matter .The earthworms of 8-10 centimeter (cm) in length and
0.2 -0.5 cm width were used for all the experiment protocol. Ready
availability, anatomical and physiological resemblance of (Pheretima
posthuma) made it to be used initially for in-vitro evaluation of
anthelmintic activity. Time for paralysis was noted either when any
movement could not be observed except when the worms where
shaken vigorously. Death was included when the worms lost their
motility followed by white secretions and fading away of their body
colour.
7/21/2022 10

11. Evaluation of Anthelmintic activity:
The antihelmintic activity was evaluated on adult
Indian earthworm. The earthworms were
randomly chosen and divided into five groups
having five earthworms in each as follows:
Group I:Ethanolic leaves extract of Tephrosia
purpurea(TPEE-25,50,100 mg/ml)
Group II: Albendazole-25,50,100 mg/ml
Group III: levamisole-5,10,15 mg/ml
Group IV: Piperazine Citrate -5,10,15 mg/ml
Group V:Ivermectin- 5,10,15 mg/ml
7/21/2022 11

12. • Observations were made for the time taken by worms to
paralyze and death was observed. Time for paralysis was
noted when no movement could be observed with a slight
pin prick method. Death was ascertained by applying
external stimuli which stimulate and induce movements in
worms as well as fade of the body color was noted.
STATISTICAL ANALYSIS:
The values are expressed as mean± SEM. The statistical
analysis was performed using one way analysis of variance
(ANOVA) followed by Dunnett’s multiple comparison test.
Comparisons were made between test and marketed
branded antihelmintic drugs. P-values <0.05 was considered
statistically significant.
The statistical analysis was done by using Graph pad prism
version no: 6.0.
7/21/2022 12

13. RESULTS AND DISCUSSION
In this study, we found that ethanolic extract of Tephrosia
purpurea possess the following chemical constituents
TABLE 1 : Phytochemical screening of ethanolic extract of
Tephrosia purpurea.
7/21/2022 13
Phytochemical
Constituents
Ethanolic extract of Tephrosia
purpurea
Alkaloids -ve
Flavonoids +ve
Carbohydrates +ve
Saponins +ve
Triterpenoids +ve
Sterols +ve
Tannins +ve
Glycosides +ve

14. 7/21/2022 14
Type of Sample Dose
(mg/ml)
Time (min) taken for
paralysis of earthworms
Mean ± S.E.M
Time (min) taken for
death of earthworms
Mean ± S.E.M
Ethanolic Extract of leaves
of Tephrosia purpurea.
25 49.000 ± 2.309 72.667 ± 0.667
50 35.667 ± 0.558 62.333 ± 0.760
100 35.667 ± 0.558 40.000 ± 1.033
Albendazole 25 80.667 ± 1.944 87.867 ± 0.860
50 31.333 ± 0.333 63.800 ± 0.970
100 15.665 ± 1.243 31.985±0.546
Levamisole 5 23.332 ± 0.146 16.833 ± 0.792
10 11.667 ± 0.558 8.416±1.056
15 7.778 ± 0.143 5.610±0.583
Piperazine citrate 5 31.584 ± 0.792 33.500 ± 0.885
10 15.500 ± 0.671 16.758±1.025
15 10.333 ± 0.870 11.166±0.732
Ivermectin 5 33.167 ± 1.014 53.333 ± 0.803
10 16.512 ± 0.546 26.666±0.762
15 11.241 ± 1.768 17.789±0.526
TABLE
2:
Anti
helminthic
activity
(time
taken
for
paralysis&
death
of
earthworm
)
by
using
EETP
&Different
marketed
branded
drugs

15. 7/21/2022 15
EETP-25mg/ml EETP-50mg/ml EETP-100mg/ml
ALBENDAZOLE-100mg/ml. Levamisole -15mg/ml Piperazine Citrate -
15mg/ml
Ivermectin -15mg/ml

16. 7/21/2022 16
FIGURE 2-Graphical representation of time taken for paralysis& death
of earthworm by using EETP &Different marketed branded drugs

17. SUMMARY AND CONCLUSION
 In the present investigation, ethanolic leaf extract of Tephrosia purpurea
posses the presence of carbohydrate, proteins, amino acids, steroids,
saponins, Flavonoids, alkaloids, tannins, glycosides and triterpenoids.
 Tannins, phenolic compounds and triterpenoids were shown to possess
anthelmintic activity. Tannins are found to bind to free proteins in the
gastrointestinal tract of the host animal or glycoprotein on the cuticle of the
parasite and cause death.
 These facts suggest that tannins present in the ethanolic extract of
Tephrosia purpurea. showed the antihelmintic effect by above mentioned
mechanisms.
 From the result shown in table-2 ethanolic extract of Tephrosia purpurea
showed anthelmintic activity in dose dependent manner giving shortest
time of paralysis and death with different marketed branded drugs.
 The study finally concluded
Levamisole>Piperazine citrate>Tephrosia purpurea leaf extract>
Ivermectin>Albendazole.
7/21/2022 17

18. BIBLIOGRAPHY
1. Modhvadia, Avdabhai R., “A Contribution to Ethnobotany of Mehsana District,
North Gujarat”, Saurashtra University, 2009.
2. S.E. Deshpande, G.B. shah and N.S. Parmar, 2003 .Antiulcer activity of Tephrosia
purpurea in rats. Indian J Pharmacol., 35: 168-172.
3. P.C. Trivedi. Ethno medicinal plants of Indian, Jaipur, India: Aavishkar Publisher
and Distributors., 2207:176.
4. S.G. Joshi Medicinal Plants. New delhi, India. Oxford and IBH Publishing Private
Limited ., 2007:211.
5. Fransworth N, R. (1990) The role of Ethopharmacology in drug
development.Nature. 787: 415-416.
6. Gupta S.S. (1994) Prospectus and Prospectives of Natural plant products in
medicine.Ind J Pharmacologic. 26: 1-12.
7. Akerele O. (1993) Natures of medicinal bounty do not throw it way, WHO
Geneva, Switzerland.
8. Tanton B. (1994) Herbal drugs and pharmaceuticals, Bisset NG Ed., (German
Edn.). CRC press.
9. Kuruvilla A. (2002) Herbal formulations as pharmacotherapeutic agents.
Indian J Exp Biol. 40(1):7-11.
10. http://www.flowersofindia.net/catalog/slides/Cardamom.html
7/21/2022 18

19. THANK YOU
7/21/2022 19