Preliminary Phytochemical Screening and Anthelmintic Activity of Acacia sinuata (Lourr.) Merr against Intestinal Helminthiasis

 

Thube Smita* , Baig R. Samrana and Gulam Mohammed Sifarish Ali

Department of Pharmacognosy, M. C. E. Society’s Allana College of Pharmacy, Azam Campus , 2390 K.B.Hidyatullah road, Camp, Pune 411 001. India

*Corresponding Author E-mail: smitathube81@rediffmail.com

 

ABSTRACT:

Extracts from the pods of Acacia sinuata (Lourr.) Merr were investigated for their anthelmintic activity against Perithima  posthuma  taking into consideration their traditional use and reported chemical constituents. Various concentrations (10-100 mg/ml) of each extract were tested in the bioassay, which involved determination of time of paralysis and time for death of the worms. The activity of methanolic and ethyl acetate extracts were found to be more significant than the standard even at a dose of 10 mg/ml. Piperazine citrate (10 mg/ml) was used as standard reference and distilled water as control. The preliminary phytochemical screening showed the presence of carbohydrates, alkaloids and saponins.

 

KEYWORDS: Acacia sinuate, Anthelmintic, Phytochemical screening, Steroids

 


INTRODUCTION:

Diseases caused by helminth parasites in livestock continue to be a major productivity constraint, especially in small ruminants in the tropics and subtropics1. In the developing world, the greatest impact of parasitic diseases is indirect and causes potential productivity losses2. Infections by gastrointestinal helminth parasites of livestock are among the most common and economically important diseases of grazing livestock3. Adulteration of anthelmintics has been found to be a common practice4. Illiteracy and unfamiliarity with synthetic anthelmintics, resulting in incorrect usage, are also a problem leading to the same consequences. Moreover, these drugs are relatively expensive. As a consequence of these problems and difficulties, pastoralists and small holder farmer have continued to use indigenous plants as livestock dewormers5. Considerable research has shown that some plants not only affect the nutrition of animals, but also have antiparasitic effects6. For example, plants that contain condensed tannins, a class of phenolic secondary metabolites, have these effects. Despite extensive use of chemicals in modern clinical practices all over the world, interest in exploiting potential of plants as source of drugs have not waned. The plants are known to provide a rich source of botanical anthelmintics, antibacterial and insecticides7. A number of medicinal plants have been used to treat parasitic infections in man and animals8,9.

 

Literature survey revealed that the extract of the pods of Acacia sinuate (Lourr.) Merr has yet not been screened for its traditional anthelmintic activity. Therefore the present study was carried out to provide pharmacological evidence for the folklore medicinal consideration of pods as anthelmintic.

 

Plant description:

Acacia sinuate (Lourr.) Merr is a common prickly, scandent shrub, with internodal thorns hooked, occurring in tropical  jungles throughout India, especially in the Deccan10. The leaves are bipinnate, pinnae 8-15 pairs, leaflets 20-25 pairs, linear, pubescent. The flowers are yellow in colour, globose with axillary heads. The fruit pods are fleshy, wrinkled when dry, depressed between seeds. Seeds are present 10-14.The plant flowers and fruits between September to March. The plant is known in the market as soap pod or soap nut11. Propogation is done by the seeds.

 

MATERIALS AND METHODS:

Plant material:

The pods of Acacia sinuate were collected in the month of October 2009 from Marketyard area, Pune. The collected pods were shade dried for 20 days and then a coarse powder was prepared. The coarse powder was stored in an air tight container and used for further successive extraction.

 

Preparation of crude extract:

On the basis of polarity different solvents like Petroleum ether, ethyl acetate and methanol were chosen for successive soxhlet extraction. Finally extracts were concentrated under reduced pressure using rotary evaporator. The percentage yield of these extracts are reported in Table I.

 

Table I Percentage yield

Sr. No

Extract

% Yield (w/w)

1.

Petroleum ether

1.1

2.

Ethyl acetate

5.5

3..

Methanol

8.2

 

Preliminary phytochemical screening:

The preliminary phytochemical screening of the plant parts was undertaken using described methods12,13.

The screening covered mainly alkaloids, glycosides, sterols, flavonoids, saponins, tannins, protein and amino acids and reducing agents. The presence of different secondary metabolites are reported in Table II.

 

Table II Preliminary phytochemical screening

Sr.

no

Test

Methanolic

extract

Ethylacetate

extract

Petroleum

ether extract

1.

Carbohydrates

 

 

 

a)

Molish

+

+

-

b)

Fehlings

+

-

-

c)

Benedicts

+

-

-

d)

Tollens

+

+

-

e)

Iodine

-

-

-

2.

Gum

-

-

-

3.

Mucilage

-

-

-

4.

Proteins

 

 

 

a)

Biuret

-

-

-

b)

Million

-

-

-

5.

Amino acid

 

 

 

a)

Ninhydrin

-

-

-

b)

Tryptophan

-

-

-

c)

Tyrosine

-

-

-

6.

Alkaloids

 

 

 

a)

Dragendorff’s

+

+

-

b)

Mayer’s

+

-

-

c)

Hager’s

+

+

-

d)

Wagner’s

+

+

-

7.

Tannins

 

 

 

a)

Ferric chloride

+

+

-

b)

Lead acetate

+

+

-

8.

Steroids

-

-

+

9.

Saponins

 

 

 

a)

Foam test

+

+

+

 

Worms collection:

Indian earthworm Pheritima posthuma (Annelida) were collected from the Government Agriculture College, Pune.

 

Preparation of test sample:

Samples for in-vitro study were prepared by dissolving the extracts in distilled water to obtain different working solutions as 10, 50, and 100 mg/ml.

 

Anthelmintic Assay:

The anthelmintic assay was carried as per the method of Ajaiyeoba E.O. et al 14 with minor modifications. The assay was performed on adult Indian earthworm, Pheretima posthuma due to its anatomical and physiological resemblance with the intestinal roundworm parasite of human beings15-18. Because of easy availability, earthworms have been used widely for the initial evaluation of anthelmintic compounds in vitro19-22. 50 ml formulations containing three different concentrations, each of crude extracts (10, 50 and 100 mg/ml in distilled water) were prepared and six worms (same type) were placed in it. Time for paralysis was noted when no movement of any sort could be observed except when the worms were shaken vigorously. Time for death of worms were recorded after ascertaining that the worms neither moved when shaken vigorously nor when dipped in warm water (50 °C). Piperazine citrate (10 mg/ml) was used as reference standard while distilled water as the control.

 

Table  III Time taken for Paralysis(P) and Death(D) of worms in minutes.

Sr.no

TEST SAMPLES

 

CONCENTRATION

(MG/ML)

TIME OF PARALYSIS (P)

DEATH

(D)

1.

Methanolic extract

10 mg/ml

50 mg/ml

100mg/ml

26 min

15 min

8 min

27 min

17 min

9 min

2.

Ethylacetate

Extract

10 mg/ml

50 mg/ml

100mg/ml

8 min

6 min

5 min

9 min

8 min

6 min

3.

Petroleum ether extract

10 mg/ml

50 mg/ml

100mg/ml

5 min

3 min

1 min

6 min

5 min

2 min

3.

Piperazine citrate

10 mg/ml

25 min

42 min

 

RESULT AND DISCUSSION:

Preliminary phytochemical screening revealed the presence of carbohydrates, saponins, and alkaloids. As shown in table III, the methanolic and ethyl acetate extract of Acacia sinuata displayed significant anthelmintic properties in a dose dependent manner giving shortest time of paralysis and death with 100 mg/ml concentrations. All the extracts not only demonstrated paralysis but also caused death of worms, even better than the standard at a concentration of 10 mg/ml. Phytochemical analysis of the crude extracts revealed the presence of saponins ( 20.8 %)which have reported to contain acacinin-C,-D and –E and alkaloids  as the chemical constituents .

 

CONCLUSION:

Results reported in the present work constitute rational evidence and a scientific basis to justify and support the folkloric claims of the potential anthelmintic activity of Acacia sinuata .Further study is required to isolate the active constituent/s for the development of a novel standardized anthelmintic herbal formulation.

 

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Received on 12.01.2011       Modified on 23.01.2011

Accepted on 31.01.2011      © RJPT All right reserved

Research J. Pharm. and Tech. 4(5): May 2011; Page 756-758