In Vitro Antihelmintic Activity and Preliminary Phytochemical Screening of Whole Plant of Vernonia cinerea

 

S. Ushasri*, K. Nagaraju, D. Prasanthi, K. Ramadevi, CH. Sudha Bhargavi.

Sri Siddahartha Pharmacy College, Ammavarithotha, Nuzvid, India-521201.

*Corresponding Author E-mail: ushasri06@rediffmail.com

 

Vernonia cinerea (Asteraceae) is a potential medicinal plant and as it is a non cultivated plant it is widely distributed in waste lands and open forests in India. The alcoholic, etheral and chloroform extracts were obtained from the roots of plant Vernonia cinerea by soxhlet extraction or continuous hot percolation methods and the extracts were evaluated for their respective antihelmintic activity, against locally available earth worms (Pheretima posthuma). Three concentrations (10, 30, 60 mg/ml) were prepared from each extract and were used for the study over earth worms. The study involves the determination of time of paralysis and time for death of the earth worms tested. The results obtained from the study revealed the fact that chloroform and alcoholic extracts from the roots of Vernonia cinerea possess significant antihelmintic effect. A solution of Albendazole was prepared in same concentration as that of the extract and was used as standard. A 2% tween-80 suspension prepared using normal saline solution was used as control or blank.

 

 

INTRODUCTION:

Vernonia cinerea (L.) belonging to the family Asteraceae is an annual plant widely distributed in India, Bangladesh, Sri Lanka and Malay island¹. It is commonly known as ‘little Ironweed’ in English. The Ayurvedha Pharmacopoeia¹ of India recommends the plant to treat intermittent fever, filariasis, blisters, boils and vaginal discharges. The roots of the plant are used traditionally for the treatment of all types of eruptive boils and the juice is used for quicker healing of accidental wounds, filariasis and toxic viral fevers. The seeds³ are used in dysuria and to treat colic in the form of decoction. The young leaves⁵ of this plant are used for the treatment of tonsillitis. The leaf juice extract is used to treat skin diseases and the leaf extract for treating dysentery in children. Besides these, the plant is used in smoking cessation, cough, fever, malaria, urinary calculi, arthritis and leprosy. The plants possess antimicrobial³, antibacterial, antioxidant, antihelmentic, anti-inflammatory, analgesic, antipyretic, antiflautulent, antispasmodic and antidiuretic⁴ properties. Some of the phytochemical compounds present are sterols, flavonoids, sesquiterpene lactones and a terpenoid, ‘leupeol acetate’ which shows antihyperglycaemic and antiulcer properties.

 

Furthermore, Vernonia cinerea also possesses insect-antifeedant property. Therefore, the present study was taken up to evaluate the crude root extracts of Vernonia cinerea for it’s against Pheretima posthuma for anti helmintic activity.

 

Helminthiasis or worm infection is one of the most prevalent diseases in the World. The conditions appear to be worsening with respect to the number of people’s affected⁵. Parasitic worm also infect livestock and crops, affecting food production with a resultant economic impact. Despite this prevalence of parasitic infections, the research on antihelmintic drug is poor⁶. Helminth infections are among the most common infections in humans, affecting a large population of the World although the majority of infections due to worms are generally limited to tropical regions and pose a great threat to health and contribute to the prevalence of malnutrition, anemia, eosinophilia and pneumonia. Parasitic diseases cause severe morbidity affecting mainly population in endemic areas with major economic and social consequences. The gastrointestinal helminthes become resistant to currently available antihelmintic drugs therefore there is a foremost problem in the treatment of helminthes diseases⁷. Therefore there is an increasing demand towards natural antihelmintics.

 

 

Fig.1. Antihelmintic activity of various extracts of Vernonia cinerea

 

Table.1. Details of the extraction of roots of Vernonia cinerea plant

Whole plant material used for the total extraction 50g	Solvent used	Color and Consistency	% yield
	Chloroform (CEVC)	Greenish black	2.54
	Ether (EEVC)	Greenish black	2.07
	Alcohol (AEVC)	Greenish black	2.89

 

Table.2. Phyto chemical constituents of various extracts of whole plant of Vernonia cinerea

S.No.	Chemical Test	Chloroform Extract	Ether Extract	Alcohol Extract
1	Alkaloids	+	+	+
2	Amino acids	+	+	+
3	Carbohydrates	+	+	+
4	Tannins	+	+	+
5	Proteins	+	+	+
6	Steroids	+	+	+
7	Flavonoids	_	+	+
8	Saponins	+	+	+
9	Glycosides	+	+	+
10	Acids	+	+	+

 

MATERIAL AND METHODS:

Collection of Plant

The leaves, stem, flowers and roots of plants Vernonia cinerea were collected from the local areas of Nuzvid in the month of August. The voucher specimen was identified with the Botanist Mr. M. Raghu Ram, Botanist in Acharya Nagarjuna University. The whole plant of Vernonia cinerea was shade dried and it was milled into coarse powder by a mechanical grinder and it was stored in a closed vessel for further use.

 

Preparation of extracts

The dried powder whole plant of about 50g placed in soxhlet extractor extracted with 500ml each of ethanol, chloroform and ether solvents respectively for 48hrs. The process was again repeated with the same quantity. The extract obtained was filtered and evaporated to dryness under reduced pressure in a rotary vacuum evaporator. The details of the extraction of plant Vernonia cinerea were presented in the table 1 and also Phyto chemical constituents in the extract were presented in the table 2.

 

Antihelmintic activity:

Chloroform, ether and alcohol extracts from the leaves, stem, flowers and roots of plant of Vernonia cinerea were investigated for antihelmintic activity against Pheretima prosthuma.

 

 

Table.3. Antihelmintic activity of various extracts of roots of Vernonia cinerea plant

Treatment	Concentration used(mg/ml)	Time taken for paralysis(min)	Time taken for death(min)
Control	Positive	-	-
	Negative	-	-
Albendazole	10	3.16±0.030	32.24±0.348
	30	3.18±0.054	30.69±0.146
	60	2.69±0.115	21.58±0.226
Chloroform	10	5.73±0.034	117.75±0.068
	30	4.70±0.023	85.43±0.086
	60	4.38±0.045	75.76±0.103
Ether	10	5.44±0.048	118.88±0.081
	30	4.61±0.037	30.69±0.146
	60	3.43±0.049	87.86±0.061
Alcohol	10	4.84±0.057	106.24±0.091
	30	4.53±0.073	84.55±0.095
	60	3.39±0.127	78.25±0.070

 

Various concentrations (10, 30 and 60 mg/ml) were prepared in 2% tween80 suspension in normal saline for each extract and were tested by bioassay, which involved determination of time of paralysis and time of death of the worms. Albendazole was used as standard reference, normal saline as positive control and 2%tween 80 in normal saline as negative control. The Antihelmintic assay was carried as per the method followed by Venkata raju et al with minor modifications⁸. The assay was performed on adult Indian earth worms, Pheretima prosthuma due to its anatomical and physiological resemblance with the intestinal round worm parasite of human beings⁹. Because of easy availability, earthworms have been used widely for the initial evaluation of antihelmintic compounds in vitro. The earthworms were collected from moist soil and washed with normal saline to remove all faecal matter and were used for the antihelmintic study. The earth worms of 8-10 cm in length and 0.3-0.4 cm in width were used for all experimental procedures. The earth worms were divided into fourteen groups containing six earth worms in each group. All the extracts and standard drug solution were freshly prepared with 2%tween80 in normal saline as suspension before starting the experiments. Different extracts and standard drug solutions were poured in different petri plates. All the earth worms were released into 1ml, 3ml and 6ml suspension which contain 10mg/ml of formulation as follows: Chloroform extract, Ether extract, Alcohol extract and Albendazole in three different concentrations. Observations were made for the time taken to paralysis and death of worms. Time for paralysis was noted when no movement of any sort could be observed except when the worms were shaken vigorously. Death was concluded when the worms lost their motility when dipped in warm water (50ºC) followed with fading away of their body colors. The results were calculated by two way ANOVA.

 

RESULTS:

Preliminary phytochemical analysis showed the presence of alkaloids, flavonoids, steroids and tannins like phytoconstituents in the whole plant extracts of Vernonia cinerea. Some of these phytoconstituents may be responsible to show a potent antihelmintic activity. From all the observations alcoholic and chloroform extracts of whole plant of Vernonia cinerea were found to show a potent antihelmintic activity when compared with the standard drug.

 

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Received on 30.11.2012         Modified on 18.12.2012

Accepted on 20.12.2012         © AJRC All right reserved