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Introduction-

Parasitic worms or Helminths are obligatory parasite found on or in the body the other organisms (hosts),viz.,human beings, domestic pets, livestokes, plants ,insects etc1.
The parts of plant Tephrosia purpurea were selected for present study
2. The well-cleaned seeds of Tephrosia purpurea Linn were powdered and extracted with three different solvents.
Adult Indian earthworms, Pheretima posthuma having anatomical and physiological resemblance with intestinal roundworm parasite of the human being. So Pheretima posthuma were used for present study2.

Objectives-

·To perform the literature review of Tephrosia purpurea
·Selection of suitable method for the evaluation of anthelmintic activity.
·Preparation of the extracts from dried and powdered stem of Tephrosia purpurea by using different solvents of different polarity.
·Pharmacognosticalevaluation of Tephrosia purpurea
· Phytochemical evaluation of different extracts.
·To make the different groups of worms.
·Preparation of test sample suspensions by using suspending agent i.e. acacia
·Determination of the anthelmintic activity of test extract and albendazole Compilation & documentation of the obtained results.


Experimental method

1.PREPARATION OF EXTRACT FOR TEST 2:
The extracts of seeds of Tephrosia purpurea Linn. were extract were suspended in 1% acacia in normal saline as a vehicle and used for anthelmintic evaluation.The Pet ether, Ethanol and Aqueous extracts of seeds of a Tephrosia purpurea Linn were suspended  at 50 and 100 mg/ml in 1% acacia in normal saline.

2.Preparation of Drug Solution:

For positive control, an Albendazole suspension was used. The selected Albendazole suspension was a marketed formulation i.e. ‘Noworm suspension’ Alkem Pharmaceutical Pvt. Ltd.

3.Selection of worm:

Adult Indian earthworms, Pheretima posthuma having anatomical and physiological resemblance with intestinal roundworm parasite of the human being. So Pheretima posthuma were used for present study.

4.Chemical used:

Drug: Albendazole                  :  ‘Noworm suspension’ Alkem Pharmaceutical Pvt.   Ltd.
                                              
Testing sample                        :  successive  pet-ether, ethanol and aqueous extracts
                                                 of seeds of Tephrosia purpurea Linn.
Vehicle                                    :     1% acacia in normal saline

5.Grouping of animals:

The experimental design of the investigation was carried out in seven groups with ten worms in each group and carried out in the following regimes.
Group I                       normal saline.
Group II                     Albendazole 25 mg/ml.
Groups III, IV           Pet-ether extracts (50,100mg/ml)
Group V, VI             
Ethanol extract (50,100mg/ml)
Group VII, VIII       
Aqueous extract (50,100mg/ml)
       

Anthelmintic Activity 3

                    Adult Indian earthworms, Pheretima posthuma was obtained and authenticated from the horticulture department of CT Bora College, Shirur. It used to screen anthelmintic activity, due to its anatomical and physiological resemblance with intestinal roundworm parasite of the human being. Eleven groups of six earthworms of approximately same size (5 to 8cm) were released in to 10ml of desired formulation at room temperature. Each of group is first treated with normal saline solution and then treated with one of the following solution. All the dried extract were suspended in 1% acacia in normal saline as a vehicle and used for anthelmintic evaluation.

The standard Albendazole (25mg /ml) and the test extract i.e. Pet- ether; ethanol and aqueous extract (50,100 mg/ml) were evaluated for anthelmintic activity. Observations were made for the time taken for paralysis and death of individual worms up to four hours of test period. The mean paralysis time and mean lethal time of each extract was recorded. Paralysis was said to be occurred when worms did not revive even in normal saline. Death was concluded when worm lost their motility followed with fading away of their body colour (Table 1)

RESULTS AND DISCUSSION

Table1. Data showing the time taken for paralysis and death by Indian earthworm
April -2009 / Volume - 1 / Article No - 5 / Research Article

“EVALUATION OF PHARMACOGNOSTICAL, PHYTOCHEMICAL AND PHARMACOLOGICAL ACTIVITY OF TEPHROSIA PURPUREA LINN”.


Miss.Surve Suvidha S
.*., Miss. Patil Anuja

Miss.Surve S.
Sr. No. Treatment Time Taken For Paralysis (Min) Time Taken For Death  (Min)
1 Vehicle    
Albendazole
2 25 mg/ml 2.26 ± 0.194 5.0 ± 0.00
3 50 mg/ml 44.0 ± 1.378 60.0 ± 0.00
4 100 mg/ml 27.0 ± 1.140 60.0 ± 0.00
5 50 mg/ml 7.6 ± 1.030 15.2 ± 1.356
6 100 mg/ml 5.4 ± 0.244 10.6 ± 0.245
7 50 mg/ml 16.2 ± 0.860 53.6 ± 2.211
8 100 mg/ml 10.4 ± 0.813 46.2 ± 3.023
(i.e. Pheretima posthuma)
Pet - Ether Extract
Ethanol Extract
Aqueous Extract
The P<0.001, considered extremely significant variation among the column. Column means is significantly greater than expected chance.

The preliminary phytochemical study investigation of all the extract shows presence of the preliminary phytochemical study investigation of all the extract shows presence of Anthroquinone.  Pet ether and ethanol extract shows presence of alkaloids, steroids and sterols, triterpenoids and fixed oil. Ethanol and aqueous extracts shows presence of flavonoids, while as aqueous extract shows presence of carbohydrate, Protein, Amino acid, tannins and phenolic compounds and saponins. Pet ether, Ethanol, Aqueous extracts of seeds of Tephrosia purpurea Linn. was evaluated for  anthelmintic activity. The observation carried out as time require for paralysis and death. The worms were considered to be paralyzed when their normal movement was changed and almost ceased, while the worms were considered to be died when their movement was completely stopped, which were further confirmed by putting them into normal saline solution.
  As shown in table1 Pet ether, Ethanol, Aqueous extracts of Seeds of Tephrosia purpurea Linn. exhibited anthelmintic activity in dose-dependent manner giving shortest time of paralysis and death with 100mg/ml concentration for Pheretima posthuma. Pet ether extract cause paralysis of 27 min and earthworm remain alive for 60 min., the aqueous extract shows paralysis within 10.4 min and time of death 46.2 min and The Ethanolic extract of Tephrosia purpurea Linn. cause paralysis within 5.4 min and time of death 10.6 min Ethanolic extract was found to be most potent among the all extract. The vehicle treated earthworm was found to be unaffected even after four hours.
The ethanolic extract shows the maximum anthelmintic activity, the different chemical present such as Alkaloids, Glycosides, Steroids and sterols, Anthraquinones, Flavonoids, Triterpenoids, and fixed oil may attribute it.
               The present study was undertaken to validate the Ethanobotanical claim of Tephrosia purpurea Linn. The result shown that the plant having potent anthelmintic activity compaired to standard drug.  In future, scientist should isolate the possible phytoconstituent, which may be responsible for the anthelmintic activity and to possible mechanism of action.

REFERENCES

1.
Goodman and Gilman’s  ‘ the pharmacological basis of therpautics ‘McGraw Hill, Edn 9th, 1996, 1009 - 1014.
2.Wealth of India, publication and information directorate, CSIR, new Delhi; Vol X, 2003, 151 - 153.
3. Turner, R. A. “Screening Methods in Pharmacology”; 3rd Edition. Academic Press Nic., London, 1965, p. 592.
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