In Vitro Licicidal Activity of Different Roots and Leaves Extract of Enicostema axillare (Gentianaceae).

 

1PB Aswar*, 1SS Khadabadi, 2BS Kuchekar ,1R Dhage and    1SP Deshmukh

1Government College of Pharmacy, Amravati 444604(M.S.), INDIA

2MIT College of Pharmacy, Kothrud Pune

 

ABSTRACT

Enicostema axillare is well known for its anthelmintic and bacterial activity. In present study dried leaves and roots of Enicostema axillare were subjected to exhaustic sequential extraction with four solvent i.e.-hexane, chloroform, methanol and distilled water respectively. All four extracts were studied for in vitro licicidal activity using goat-lice Damalinia caprae (Trichodectidae) as experimental organism. Only n-hexane and chloroform fractions showed statistically significant licicidal activity (p value <0.001, n = 5). Decrease in mean time required to kill the lice was observed at concentration 1% w/w and 10% w/w when compared to 1% lindane solution.

 

KEYWORDS: Enicostema axillare leaves and roots, licicidal activity.

 

 

INTRODUCTION:

Enicostema axillare (Gentianaceae) in India commonly known as nai. It has been used in the indigenous system of medicine for, acrid, thermogenic, canninative, and laxative and as an anthelmintic3. Phytochemical review shows the presence of tannins, resins and starch, ophelic acid, chiratin, in the roots and leaves of Enicostema axillare . Various Pharmacological activities have been reported with Enicostema axillare.

 

The aim of present work was to evaluate licicidal activity of different extracts obtained by exhaustive sequential extraction of Enicostema axillare root and leaves by using in vitro method. Feeding habit and anatomy of goat lice Damalinia caprae closely resemble human head-lice Pediculus humanus (capitis) hence goat lice was selected and used as experimental organism. 1% lindane topical lotion is commonly used as a synthetic insecticide to treat lice infestation hence licicidal activity of Enicostema axillare was compared with lindane.

 

MATERIALS AND METHOD:

Plant Material:

The plant of Enicostema axillare was collected from Amravati (Maharashtra) in the month of August and authenticated by Prof.Marathe, Vidarbha Institute of Humanities and Science, Amravati (Maharashtra). The plants species were cultivated and preserved in the institute as a herbarium.

 

Extraction:

The plant material (leaves and roots) were dried for several days under shadow and powdered with the help of an electric grinder. It was air dried and exhaustively extracted in soxhelet apparatus successively by using chloroform, n-hexane, methanol and distilled water. Each time marc was dried at 45˚C and solvent was evaporated at 45˚Cto get respective extract.

 

Finally the dried marc was boiled with distilled water for 30 minutes, filtered and filtrate was evaporated at 45˚C to obtained solid aqueous extract.


Table 1: Effect of Enicostema axillare on goat -lice Damalinic caprae .

Group

Treatment

Mean Time in Minutes ± S.E.M.

Group 1

0.1 % w/w n-Hexane extract  test solution

154* ± 1.87

Group 2

1 % w/w n-Hexane extract test solution

66* ± 3.00

Group 3

10 % w/w n-Hexane extract test solution

34* ± 3.00

Group 4

0.1 % w/w Chloroform extract test solution

168* ± 2.55

Group 5

1 % w/w Chloroform extract test solution

84* ± 1.87

Group 6

10 % w/w Chloroform extract test solution

46* ± 2.24

Group 7

0.1 % w/w Methanol extract test solution

255* ± 1.87

Group 8

1 % w/w Methanol extract test solution

No Effect on motility and mortality

Group 9

10 % w/w Methanol extract test solution

No Effect on motility and mortality

Group 10

0.1 % w/w Aqueous extract test solution

No Effect on motility and mortality

Group 11

1 % w/w Aqueous extract test solution

No Effect on motility and mortality

Group 12

10 % w/w Aqueous  extract test solution

No Effect on motility and mortality

Group 13

1 % w/w Lindane solution ( standard )

94* ± 5.25

n = 5 in each group. Values are Mean time in minutes at which lice were considered dead ± S.E.M. *p value < 0.01

 

 


Test Solutions:

0.1,1,10 % w/w of test solutions of n-hexane extract and chloroform extract were separately prepared in coconut oil. 0.1, 1, 10 % w/w of test solutions of methanol extract and aqueous extract were separately prepared in distilled water.

 

Standard solution:

1% w/w lindane solution.

 

Experimental Organism:

Goat-lice Damalinia caprae (Trichodectidae) were collected from healthy goat located in Amravati (MS).In an earlier observation independent of present study the lice were found to remain live for 24-48 hours when removed and kept away from host body.

 

Experimental procedure:

This was carried out by modifying a method used by Pollack et al. (1999). A 25 ml capacity glass beaker was taken. A filter paper disc coinciding with internal diameter of the beaker was cut .0.15 ml test solution was applied as thin layer on filter paper disc at the bottom of beaker. A group of 5 lice was placed over the disc and observed for motility and death time through magnifying glass. The time in minute at which each lice became immobilize was noted down.

 

The immobilized lice were taken out, placed on on fresh dry filter paper and observed for 6 hours at interval of 30 minutes. The lice which did not show movement during this period were considered dead at the time when these were became immobile. Same procedure was carried out for each test solution and standard solution.

 

Statistical Analysis:

Separate group of 5 lice was assigned to each test solution and standard drug hence n = 5. All the value expressed as Mean ± S.E.M. The data was analysed by test. P value < 0.01 was considered statistically significant.

 

RESULTS AND DISCUSSION:

Methanol and aqueous extracts not showed licicidal activity. Values of mean time in minute at which lice became immobilized and considered dead on treatment with n- hexane extract, chloroform extract and standard drug lindane are given are given in table 1.Both n-hexane and chloroform extracts showed significant decrease in the mean time required to kill lice with 1% and 10% concentration where as increase in the mean time was observed with 0.1 % concentration when compared to 1% lindane. n-hexane extracts at all concentration 0.1,1 and 10%w/w.Thus from the present study it is evident that leaves and roots of Enicostema axillare contain chemical constituents responsible for licicidal property and the constituents are predominately soluble in n-Hexane.

 

In future by using n-hexane and chloroform leaves and roots extracts of  Enicostema axillare  herbal lotions or shampoos can be formulated which will be substitute for traditional chemical and synthetic antilice formulations available in market which may be economic and side effects free. As the formulations available in markets showed various side effects and adverse reactions and expensive.

 

 

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Received on 02.09.2009

Accepted on 07.10.2009     

© A&V Publication all right reserved

Research Journal of Pharmacognosy  and Phytochemistry. 1(3): Nov. – Dec. 2009, 238-239