Phytochemical Screening and Invitro Anthelmintic Activity of a Novel Polyherbal Formulation

 

Fouzia Tehseen1*, Syed Safiullah Ghori2, Mohammed Dawood1, Shafia Tazeen1, Kulsum Sara1, Abdul Samad3

1Department of Pharmaceutical Chemistry, Anwarul Uloom College of Pharmacy,

Hyderabad, Telangana, India, 500008.

2Department of Pharmacology, Anwarul Uloom College of Pharmacy, Hyderabad, Telangana, India, 500008.

3Research Scholar, Government Nizamia Tibbi College, Charminar, Hyderabad, Telangana, India, 500008.

*Corresponding Author E-mail: fouziatehseen@gmail.com

 

ABSTRACT:

Ayurveda is the ancient Indian system of medicine which includes the usage of natural and mechano procedures to eliminate the root cause of the disease. Ayurvedic system of medicine is economical, easily available and free of side effects which make it beneficial to human beings. Among various diseases, a large population is infected with helminthiasis which can be treated by using medicinal plants. In the present study we developed a novel polyherbal formulation (PHF) containing a mixture of powdered Artemesia absinthium (shoot), Butea Monosperma (leaves), Ipomea headeraceae (seeds), Hyocyamus niger (seeds). Phytochemical screening of the extract indicated the presence of carbohydrates, amino acids, tannins, sterols and alkaloids the polyherbal formulation was studied for its anthelmintic activity using Indian earthworm (Pheritima posthuma) as the test model. Albendazole was taken as the standard reference drug. The petroleum ether and aqueous extracts with the petroleum ether and methanolic extract. of the PHF of different concentrations (25µg/ml, 50µg/ml, 100µg/ml) were prepared and used to determine the paralysis time and death time of the worms. The petroleum ether and methanolic extract of the PHF has shown significant activity due to presence of various phytoconstituents like tannins, flavonoids and alkaloids. In the course of our investigation, we found that the 100µg/ml methanolic extract has better anthelmintic activity when compared to the other concentrations checked.

 

KEYWORDS: Anthelmintic drug, Polyherbal formulation, Earthworm, Albendazole.

 

 


INTRODUCTION:

The ayurvedic system was first developed between 2500 and 500 BC in India1. Ayurveda literally means “science of life,” as Indian health care management is based on the man and his illness. Medicinal plants are used as alternative or traditional drugs, neutraceuticals, diet supplements and as alternative and complimentary medicine2. WHO report shows that 80% of world population is dependent on the herbal drugs3-5. Helminths such as pin worm, ring worm, tape worm, hook worm and others are major cause of acute and chronic infections in humans.

 

Helminthiasis is prevalent globally but more cases are reported in developing countries including India. Higher prevalence when compared to developed countries is due to poor personal and environmental hygiene and cleanliness. Since ancient times medicinal plants have been used as a cure for various ailments. These plants are used effectively for the treatment of helminthiasis. Helminths harm the host by depleting the food, blood loss, pneumonia, eosinophilia, obstruction of intestines and lymphatic system and may secrete toxins6. Anthelminthic are the drugs which expel the parasitic worms from the host either by killing (vermicides) or stunning (vermifuge)7. The resistance of the worm to suggested dose and treatments is a major drawback to the control of worm in future. Helminths that are resistant to the drugs, survive and pass their resistance genes. This may result in failure of treatment if resistant worms’ genes accumulate. Intestinal worm infections can be treated more effectively than other locations in the body8. Earlier ancient system of medicine reports shows excellent anthelmintic activity9. Drugs derived from plants are highly suggestive as they are economic, easily available, safer nd nontoxic with an altered site of action10. The present work conceived by us to evaluate anthelmintic activity of a novel Polyherbal formulation has Artemisia absinthium, Butea monosperma, Ipomea hederacea and Hyocyamus niger belonging to the families Asteracea, fabaceae, Convulaceae and Solanaceae respectively.

 

MATERIAL AND METHODS:

Plant material collection:

The raw plant material namely Artemisia, Butea, Ipomea and Hyoscyamus were purchased from local market in Hyderabad. These were identified and authenticated by Dr. Muhd Ghouse, Almo Herbal Health Care, Abids, Hyderabad, Telangana.

 

Preparation of the PHF:

All the plant materials were grounded in a grinder separately and mixed in proportions given in the table 1 and stored in air tight container. About 50 gms of the PHF was subjected to continuous soxhlet extraction using Petroleum ether and methanol. (Table No 1)

 

Phytochemical Screening:

The PHF extracts were screened to check the presence of phytochemicals using standard qualitative methods11. Tests for alkaloids, carbohydrates, flavonoids, amino acids, tannins, saponins and glycosides were done on both extracts.

 

Evaluation of In vitro Anthelmintic Activity:

The invitro anthelmintic activity of the PHF was evaluated on Indian earthworm (Pheritima posthuma) due to its anatomical and physiological resemblance with the intestinal round worm parasites in humans. The modified method developed by Ajaiyeoba et.al, (2001) was used for the assay12-14. Albendazole (15µg/ml) was used as reference standard drug and distilled water as vehicle control. For the study, petroleum ether (PHFP) and methanolic extracts (PHFM) of the PHF as different concentrations of 25µg/ml PHFP1, 50µg/ml PHFP2, 100 µg/ml PHFP3, 25µg/ml PHFM4, 50µg/ml PHFM5 and 100µg/ml PHFM6 were prepared using distilled water. The selected earthworms were divided into nine groups each containing six earthworms contained in petridishes. Freshly prepared PHF solutions with different concentrations were used to record the time of paralysis when the earthworm did not show any sort of movement even after exposing to external stimuli such as normal saline. The time of death was noted when earthworms did not move even after shaking vigorously nor when placed in warm water and their body colour faded.

 

RESULTS AND DISCUSSIONS:

The phytochemical screening done on the PHF showed the presence of carbohydrates, tannins, sterols, alkaloids and flavonoids (Table 2). The Polyherbal formulation of petroleum ether (PHFP) and methanol (PHFM) showed significant anthelmintic activity. The PHF extracts are better anthelmintic when compared to Albendazole. It was found that the PHFM at 100µg/ml concentration was more effective than the other concentrations. The result of the anthelmintic activity is shown in Table 3 and figure 1. It can be observed that activity is dose dependent and is inversely proportional to the time of paralysis and time of death. Albendazole causes flaccid paralysis on the earthworm which results in expulsion of the worm by peristalsis. Moreover, albendazole brings about increased chloride ion conductance in the muscles of worms producing hyperpolarization and reduced excitability resulting in muscle relaxation and flaccid paralysis. The exact mechanism of action of the PHF cannot be clearly shown based on our results. However, it can be expected due to the depletion of glycogen synthesis or inhibition of glucose uptake in the parasites. It may have activated nicotinic receptor in the earthworms resulting in persistent depolarization or hyperpolarization15.


 

Table 1: Composition of the Polyherbal formulation

S. No.

Botanical Name

Common Name

General Name

Family

Plant Part Used

%w/w in grams per 100 grams

1

Artemesia absinthium

Worm wood

Vilayati afsantin

Asteraceae

Aerial shoot

30

2

Butea monosperma

Flame of the forest

Kamarkas

Fabaceae

Leaves

25

3

Ipomea hederaceae

Ivy leaves

Kaladana

Convulvulaceae

Seeds

25

4

Hyocyamus niger

Henbane

Ajwain

Solanaceae

Seeds

20



Table 2: Phytochemical Screening of Polyherbal Formulation

S. No.

Name of Test

Phytoconstituent

Petroleum ether extract (PHFP)

Methanolic extract (PHFM)

1

Dragendroffs Test

Alkaloids

+

+

2

Benedicts Test

Carbohydrates

-

+

3

Ninhydrin Test

Amino acids

-

-

4

Sulfuric acid Test

Flavonoids

+

+

5

Raymonds Test

Glycosides

-

-

6

Lead acetate Test

Tannins

-

+

7

Salkowski Test

Sterols

+

+


Table 3: Anthelmintic Screening of the Polyherbal Formulation

Group S. No.

Treatment

Chemical concentration (µg/ml)

Time of Paralysis (MIN)

Time of Paralysis (MIN)

1

Distilled water

-

-

-

2

Albendazole

15

25

38

3

PHFP1

25

20

26

4

PHFP2

50

14

23

5

PHFP3

100

12

20

6

PHFM1

25

18

24

7

PHFM2

50

15

17

8

PHFM3

100

13

15

 

Figure 1: Effect of Albendazole, Petroleum ether extract (PHFP) and Methanolic extract (PHFM) of the Polyherbal Formulation on Indian Earthworm (Pheritima posthuma).

 

CONCLUSION:

Medicinal plants are utilized since ancient times for the treatment of diseases without knowledge of the exact mechanism of action of the plant. However, with the recent advancements in the research studies, we can assume the possible action of the plants. The synthetic available drugs for the treatment of helminthiasis are more costly with side effects when compared to the medicinal plants which are economic, easily procurable and have fewer side effects on the host. The PHF under study has significant anthelmintic activity due to the synergistic effect of the plants used. All the PHF extracts showed good vermifugal and vermicidal activity when compared to the standard reference drug. The methanolic PHF at concentration 100µg/ml has shown to the best activity. Hence the PHF can be further studied and modified to be an effective anthelmintic drug.

 

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Received on 21.05.2020           Modified on 25.06.2020

Accepted on 29.07.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(5):2742-2744.

DOI: 10.52711/0974-360X.2021.00483