In vitro Antihelmintic Evaluation of a Novel Polyherbal Formulation

Dr. Fouzia Tehseen^1*, Dr. Syed Safiullah Ghori¹, Ruqya Khatoon³, Jabeen Sultana³

Md Sadath Ali³, Maria Mahveen³

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

²Department of Pharmacology, Anwarul Uloom College of Pharmacy, Hyderabad

³Students, Anwarul Uloom College of Pharmacy, Hyderabad

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

ABSTRACT:

Ayurveda is a holistic traditional system of medicine in India which was established centuries ago. Ayurveda is also known as “Mother of all healing” Drug formulations in Ayurveda can be of two types- single drug formulation and polyherbal formulation where a mixture of powdered drugs are used to bring about synergistic effect. One of the diseases of concern for human beings is helmintic infections. A large population in developing countries is affected is a variety of helminth parasites like hook worm, tape worm and others. We have developed a novel PHF containing a mixture of powdered bachhi, bael, katha, guggul, spogel and vidanga. The objective of the present study is to formulate and evaluate the antihelmintic activity of aqueous, methanol and petroleum ether extract of a PHF containing a mixture of powdered ayurvedic plants in earthworm, Pheritima posthuma which bears anatomical resemblance with the parasitic helminths.

KEYWORDS: Ayurveda, antihelmintic activity, polyherbal formulation.

INTRODUCTION:

Ayurvedic medicine, an ancient vedic knowledge is considered to be one of the oldest healing sceinces and is popular in present generation over many centuries of tradition. It was first originated in India thousands of years ago. Ayurveda is known as the “Mother of all healing¹”. Etymologically speaking, it is the combination of the Sanskrit words ayur (life) and veda (science or knowledge), which means “the science of life,” which focuses on bringing balance and harmony in al areas of life including mind, body and spirit². Historically, the Ayurvedic literature “Sarangdhar Samhita” dated centuries ago in 1300 A.D. has polularies the concept of polyherbalism in the ancient medicinal system³.

Traditionally the plant formulations and combined extracts of plants are taken rather than individual drugs. It is known that Ayurvedic herbals are prepared in various dosage forms and are mostly PHF^4,5.

Drug formulation in Ayurveda is based on two principles:

1. Single drug.

2. Polyherbal formulation (PHF) which employs use of more than one drug.

This traditional therapeautic herbal strategy involves combining several medicinal herbs to produce synergistic effect, also known as polypharmacy or polyherbalism.

PHF- Poly herbal formulation involves the use of more than one herb in a medicinal preparation. This concept is found in traditional medicinal systems where a combination of herbs in particular ratio are used in the treatment of illness⁶. Plant formulations and combined extracts of plants are chosen over single drugs forms.

Present work:

One of the most common infections in human beings is helmintic infections which effect a large polulation. The developing countries are at high risk to helmints and the infections due to prevalence of malnutrition, unhealthy cleanliness resources and unhygienic food habits. Most prevalent intestinal helminths are tape worm(Teania solium), round worm (Ascaris lumbricoides), book worm (Ancylostoma duodenale) and others. Risk of diseases like schistosomiasis, filariasis and onchocerciasis is major affecting large populations in the tropical developing countires including India. The traditional plants and herbs are used as medicines as they are potentially useful showing therapeutic effect. The newer drugs of allopathy are costly and less available in remote populations. These drugs have side effects like nausea, giddiness etc which limits its use over herbal drugs. The antihelmintic herbal drug should be effective with broad spectrum of action against worms. As single dose with no toxicity to the host with cost effectiveness many herbs. Polyherbal formulations are popular and highly effective with potential results. In vitro antihelmintic are performed using adult earthworm due to its physiological and analytical resemblance to the parasital intestinal worms found in human beings.^7-10

MATERIALS AND METHOD:

The objective of this study is to evaluate the synergistic antihelmintic activity of aqueous, methanol and petroleum ether extract of a PHF containing a mixture of powdered ayurvedic plants in earthworm, Pheritima posthuma. Our PHF ingredients used are as follows

Table1: Ingredients/Composition of PHF

S.No.	Common Name	Botanical Name	Family	% W/W IN 100 GMS.
1	Babchi	Psoralea Corylifolia	Fabaceae	35
2	Bael	Aegle Marmelos	Rutaceae	20
3	Katha	Acacia Catechu	Fabaceae	20
4	Guggul	Balsamodendron Mukul	Burseraceae	10
5	Spogel	Plantago Ovata	Plantaginaceae	10
6	Vidanga	Embelia Ribes	Myrsinaceae	5

Collection and authentication:

The powdered drug was purchased authenticated by Dr. Muhd. Ghouse, Almo Herbal Health Care, Abids, Hyderabad, Telangana.

Preparation of extract:

About 50 gms of powdered drug in ratio mentioned in table was extracted by successive solvent extraction process.

Phytochemical screening was performed for the detection of phytoconstituents like carbohydrates, tannins, sterols, amino acids, glycosides etc.For the study 5 mg/ml and 10 mg/ml extracts of water, methanol and petroleum ether of the PHF were tested for parameters like time of paralysis and time of death.

Drugs and Chemicals:

Albendazole, 200 mg, Ranbaxy, New Delhi was used as reference standard.

Normal saline was used as control for the study.

In Vitro Antihelmintic Activity:

The antihelmintic activity was performed on adult earthworms due to its anatomical and physiological resemblance to the round worm found as parasite in human beings using the standard procedure given. Nine groups of almost equal sized earthworms consisting of about 4 worms were exposed to 10 ml of desired PHF concentration solution.

Each group was exposed to normal saline (control), albendazole (5mg/ml and 10 mg/ml), PHF (aqueous, methanolic and pet ether each with mg/ml and 10 mg/ml).

Observations were made for the paralysis time and time of death of the individual group of worms. Paralysis occurs when the worms did not revive even in normal saline.

Death was supposed to occur on long standing no revive of the worms even after saline treatment.

Fig1: Nine groups of earthworms containing saline, albendazole, PHF with aqueous and methanol and petroleum ether with 5 and 10 % concentration.

RESULTS AND ANALYSIS:

Table 2: Phytochemical Investigation

S. No	Phytochemical Investigated	Name of Test	Methanol Extract	Petroleum Ether Extract	Aqueous Extract
1	Carbohydrates	Benedicts Test	+ VE Green colour ppt.	-VE No change	+VE Dark Green ppt.
2	Sterols	Salkowski Test	+ VE Chcl3 layer shows red colour and acid layer is transperant	+ VE	+VE
3	Amino Acid	Ninhydrin Test	-VE No purple blueish color	-VE	-VE
4	Flavanoids	Sulfuric Acid Test	+VE for Chalcones. Dark Red is Seen	+Ve for Flavanone. Orangish red Color Seem	+ Ve for Chalcones. Dark Red Seen.
5	Glycosides	Raymonds Test	-VE	-VE	-VE
6	Tannins	Lead Acetate Test	+VE Milky White PPT	-VE No Change	+VE Milky White PPT

From the in vitro antihelmintic test results it can be observed that the methanolic extract is most potential when compared to other treatment on the earthworms. The PHF drug is found to be highly potential when compared to albendazole. The time required for paralysis was found to be inversely proportional to time.

There is no clear evidence of mechanism of action of the PHF. We can expect the paralysis and subsequent death due to depletion of glucose avalability which is primary source for the ATP synthesis in the worm. It can also be suggested that paralysis may be due to the nicotinic or muscuranic receptor activation.

Table 3: Antihelmintic activity of PHF

S. No. Grp.	Treatment	Time of Paralysis	Time of Death
1	Normal Saline	-	`
2	Albendazole 5 %	14	18min
3	Albendazole 10%	15 min	19 min
4	Aqeous Extract Of PHF 5%	10min 30 sec	15 min 38 sec
5	Aq. Extract of phf 10%	6min 10 sec	12min
6	Methanolic Ext Of PHF 5%	52 sec	1.5 min
7	Methanolic Ext Of PHF 10%	5 sec	50 sec
8	Pet.Ether Ext Of PHF 5%	52sec	1 min 5 sec
9	Pet. Ether ext of PHF 10%	10 sec	2 min

CONCLUSION:

From the present investigation we can conclude that the PHF is highly potent showing synergistic effect and can be used for treatment of various helminths. The short paralysis and death of worms suggests that the drug can be used effectively in small doses for the relief from helminths.

REFERENCES:

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7. Vidyarthi RD, A Textbook of ZooLogy, S. Chand and co., New Delhi, 14^thedition 1967, pp 329-370.

8. Vigar Z, Atlas of Medical Parasitology, P.G Publishing House, Singapore, 2^ndedition, 1984, pg 216.

9. Chatterjee KD, Parasitology, Protozoalogy, and Helminthology, Guha ray Sree Saraswathy Press Ltd, Calcutta, 1967, PP 168-169.

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Received on 02.07.2019 Modified on 08.08.2019

Research J. Pharm. and Tech 2020; 13(2):636-638.

DOI: 10.5958/0974-360X.2020.00121.3