In-Vitro Evaluation Of Smilax Zeylanica Linn. Leaves For Anthelmintic Activity
V RAJESH, P PERUMAL, V CHINTHAKINDHI, S PRABHAKARAN, G HYMAVATHI, T GUNTUPALLI
Keywords
anthelmintic, pheritima posthuma, smilax zeylanica
Citation
V RAJESH, P PERUMAL, V CHINTHAKINDHI, S PRABHAKARAN, G HYMAVATHI, T GUNTUPALLI. In-Vitro Evaluation Of Smilax Zeylanica Linn. Leaves For Anthelmintic Activity. The Internet Journal of Pharmacology. 2009 Volume 9 Number 1.
Abstract
The objective of the present study was to evaluate the
Introduction
Helminth infections are among the most common infections in man, affecting a large proportion of the world’s population. Parasitoses have been of concern to the medical field for centuries and helminths still cause considerable problems for human beings and animals. During the past few decades, despite numerous advances made in understanding the mode of transmission and the treatment of these parasites, there are still no efficient products to control helminthes and the indiscriminate use of some drugs has generated several cases of resistance. Furthermore, it has been recognized recently that anthelmintic substance having considerable toxicity to human beings are present in foods derived from livestock, posing a serious threat to human health. Consequently, the discovery and development of new chemical substances for helminth control is greatly needed and has promoted studies on traditionally used plants which is generally considered to be very important source of bioactive substances.
It was observed from ayurvedic literature and ethanobotanical studies that the plant
The literature survey revealed that no systematic studies were carried out to evaluate the anti parasite potential of
Materials and Methods
Drugs and Chemicals
The following drugs and chemicals were used
Drug: Albendazole (Kemwell Pvt.Ltd, Bangalore).
chemicals: Petroleum ether AR (60-800C) Merck Specialities Pvt. Ltd.), Benzene AR (Merck Specialities Pvt. Ltd.), Chloroform AR (Nice Chemicals Pvt. Ltd.), Methanol AR (Thomas Baker Chemical Pvt. Ltd.), Tween 80. All other reagents used were of analytical grade.
Plant Material
The fresh leaves of
Extraction
The powdered leaf material was successively extracted by using soxhlet extraction. Solvents were used with increasing polarity: Petroleum ether, benzene, chloroform and methanol. The extracts were evaporated to dryness and phytochemical screening were performed1,3.
Animals: Swiss albino mice of female sex weighing 20-25gms were employed for toxicity study. . They were housed in standard environment condition and fed with standard rodent diet with water and
Toxicity Study
An acute oral toxicity study was performed as per OECD guidelines 423 by Acute toxic class method4. Swiss albino mice of female sex weighing 20-25gms were used for the study. Acute toxic class method is a stepwise procedure with use of three animals of a single sex per step¬. Depending on mortality or morbidity status of the animals, an average of 2-4 steps may be necessary to allow judgement on the acute toxicity of the substance. Three animals were used for each step. The animal were placed individually and observed for any sign of toxicity, morbidity or mortality during the first 24hrs, with special attention given during the first 4 hours and daily thereafter for a total of 14 days.
Evaluation of
Indian adult earthworms
All the leaf extracts (Petroleum ether extract, benzene extract, chloroform extract and methanol extract) were evaluated for anthelmintic activity. Two doses of extract 20mg/ml and 40mg/ml were selected to evaluate the anthelmintic activity. The drug albendazole 20mg/ml was used as reference standard. The worms of equal size were divided into twelve groups containing six worms in each group. All the extracts and the standard drug solution were freshly prepared before starting the experiment. The extracts were dissolved in minimum quantily of Tween80 and the volume was adjusted to 10ml with saline water to desired concentrations (20mg/ml and 40mg/ml). Standard drug solution was prepared freshly before experiment. Different extracts and the standard drug solution were poured in petridishes labelled with extract and concentrations. Tween 80% in normal saline, volume adjusted to 10ml was used as vehicle control.
Six worms of equal size were introduced into each Petridish and time was noted. Observations were made for paralysis and death. The mean time of paralysis and death was recorded in minutes. The paralysis time was recorded when no movement of any sort could be observed except when the worms were shaken vigorously. Time for death of worms was recorded when worms neither moved while shaken vigorously nor when dipped in warm water (500C) with fading away of their body colors.
Statistical Analysis
All the grouped data were expressed as mean
Result and Discussion
The percentage yield of Petroleum ether, Benzene, Chloroform and Methanol extract was found to be 2%, 1%, 1% and 19.63%.
The preliminary phytochemical analysis showed the presence of alkaloids, phytosterols in all the four solvent extracts (Pet ether, Benzene, Chloroform and methanol). The presence of Flavonoids and carbohydrates were noted in methanol extract and terpenoids in chloroform extract.
The acute oral toxicity of various extracts of
Figure 1
All the value are expressed as mean
From the observations made which is shown in table 1, the petroleum ether extract of
The Benzene extract of
The chloroform extract of
The methanol extract of
From the results, it is clear that petroleum ether and chloroform extract showed potent anthelmintic activity than standard drug Albendazole. The Benzene extract was inactive at 20mg/ml concentration but showed a dose dependent action on paralysis and death.
The methanol extract showed a dose dependent anthelmintic activity. The time taken for death of worms were least compared to albendazole.
The findings suggest, the difference in anthelmintic activity between extracts is due to presence of active constituents responsible for anthelmintic activity
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
5 A – 20mg/ml
5 B – 40mg/ml
Figure 7
6A – 20mg/ml
6B- 40mg/ml
Figure 8
7 A – 20mg/ml
7 B – 40mg/ml
Figure 9
8 B – 40mg/ml
8 A – 20mg/ml
Conclusion
From the results, it is concluded that the extracts of leaves of
Acknowledgement
The authors are thankful to the management of JKK Nataraja College of Pharmacy, komarapalayam, Tamilnadu, INDIA for providing necessary facilities to carry out the research work.