In- Vitro Anthelmintic Evaluation of Polyherbal Formulation: Krumighattini Tablet

Shahin Khan¹, Ajay Kumar Meena², Ajay Saluja¹

¹A. R. College of Pharmacy & G. H. Patel Institute of Pharmacy, Anand

²Central Council for Research in Ayurvedic Science, Janakpuri, New Delhi

*Corresponding Author E-mail: inshah01@rediffmail.com, ajaysheera@gmail.com

ABSTRACT:

Ayurveda has abundant collection of effective formulations against diseases. The polyherbal formulation contains Vernonia anthelmintica, Curcuma longa, Piper longum, Mallotus philippinensis, Operculina turpethum, Terminalia chebula, Butea monosperma, and Red ochre. The present study deals with anthelmintic evaluation of ayurvedic formulation. Various concentrations of polyherbal formulation (10 mg/ml, 20 mg/ml and 30 mg/ml) in 50 ml of normal saline were taken. The results were compared with standard albendazole (10mg/ml, 20mg/ml and 30 mg/ml) and control was taken as normal saline (0.9% NaCl). The results were expressed in terms of time in minutes for the paralysis and time of death of the worms. Paralysis and death time were analyzed using one way ANOVA analysis using Graph pad prism 2.01 Software. The study indicates that polyherbal formulation shows more potent anthelmintic activity than standard reference.

KEYWORDS: Polyherbal formulation, Krumighattini tablet, anthelmintic activity, albendazole.

INTRODUCTION:

Herbal medicines are in great demand in the developed as well as developing countries for primary healthcare because of their wide biological activities, higher safety margins and lesser costs ^[1]. Ayurveda can be considered as the treasure of India having abundant formulation for various diseases. During the past decade, there has been increasing acceptance as well as public interest in natural therapies in both developing and developed countries ^[2]. Ayurveda gives various formulations for the treatment of helminthiasis.

Helminthiasis is a macroparasitic disease of humans and animals in which a part of the body is infested with parasitic worms such as pinworm, roundworm or tapeworm. More than half of the population of the world suffers from infection of one or the other and majority of cattle’s suffers from worm infections ^[3]. Anthelmintics are drugs that expel parasitic worms from the human body ^[4]. Treatment with an anthelmintic drug kills worms whose genotype renders them susceptible to the drug. Worms that are resistant survive and pass on their “resistance” genes. Resistant worms accumulate and finally treatment failure occurs ^[5].

Indiscriminate uses of synthetic anthelmintics can lead to resistance of parasites. Herbal drugs have been in use since ancient times for the treatment of parasitic diseases in human and could be of value in preventing the development of resistance ^[6].The objective of this study was to investigate the anthelmintic activity of krumighattini tablets. The polyherbal formulation contains Vernonia anthelmintica, Curcuma longa, Piper longum, Mallotus philippinensis, Operculina turpethum, Terminalia chebula, Butea monosperma, and Red ochre.

MATERIALS AND METHODS:

Krumighattini tablets were prepared in laboratory by taking the same ingredients as mentioned above. Sodhana of Mallotus philippinensis and Red ochre were done as mentioned in Ayurvedic formulary. Dry granulation method was followed. Fine powder of each ingredient was passed through sieve no. 80 and compressed to form tablets using binding agent.

Collection of Worms:

The Indian Earthworms Pheretima posthuma were collected from Anand Agricultural University Farm and authenticated from the Department of Zoology from V.P. Science College, Vallabh Vidhyanagar. It was washed with normal saline to remove all fecal matter can be used for the anthelmintic study. The earthworms of 3-5 cm in length and 0.1-0.2 cm in width were used for all the experimental protocol. Ascardia species worms are easily available in plenty from freshly slaughtered fowls and their use, as a suitable model for screening of anthelmintic drug was advocated earlier. It was collected from the freshly slaughtered Fowl intestine and washed with Normal saline to be used for anthelmintic activity ^[7,8].

In vitro anthelmintic activity:

The anthelmintic assay was carried out as per the method of Ajaiyeoba et al. ^[9] with minor modification. The assay was performed on adult Indian earth worm Pheretima posthuma due to its anatomical and physiological resemblance with the intestinal roundworm parasite of human beings^[10]. Three groups of treatment, each consisting of three worms (Pheretima posthuma and Ascardia species) in each treatment from each model were released into 50 ml of herbal tablet formulation (10 mg/ml, 20 mg/ml and 30 mg/ml) and compared with standard Albendazole with same concentration. Observations were made on the basis of the time taken for paralysis and/or death of worms. Time for death of worms were recorded after ascertaining that worms neither moved when shaken vigorously nor when dipped in warm water (50^οC). Three replications of each treatment should be maintained to estimate any sources of error. Paralysis was said to occur when they did not revive even in saline water. Death was concluded when the worms lost their motility followed with fading away of their body color.

Statistical Analysis:

Results are expressed as Average± S.E.M of three worms in each group for paralysis and death time using one way ANOVA analysis using Graph pad prism 2.01 software.** symbol represents the statistical significance p value; *p<0.01, **p<0.001 comparison are made with standard group albendazole ^[11].

RESULTS AND DISCUSSION:

The evaluation of anthelmintic activity on earthworm revealed that the activity was dose dependent and inversely proportional to paralysis and death time. From the graph (Figure 1 and 2), it is clearly seen that the herbal tablet is more potential than reference control. The activity of herbal tablet is more significant at 10mg/ml with significant level **p< 0.001.

Table 1. In vitro anthelmintic activity of herbal tablet on earth worms (Pheretima posthuma)

Groups	Treatment	Dose (mg/ml)	Paralysis time	Death time
I	Normal saline	--	--	--
IIa	Albendazole	10	54±0.57	66±0.57
IIb		20	34±0.57	41.33±0.88
IIc		30	21.67±0.88	32±0.57
IIIa	Anthelmintic Herbal tablet	10	41±1.155	53.6±0.88
IIIb		20	31.667±0.667	38.6±0.88
IIIc		30	21±1.155	28.66±1.453

Table 2. In vitro anthelmintic activity of herbal tablet on round worm (Ascaridia species)

Groups	Treatment	Dose (mg/ml)	Paralysis time	Death time
I	Normal saline	--	--	--
IIa	Albendazole	10	38.667±0.33	57.33±0.67
IIb		20	22±0.57	39.83±0.60
IIc		30	12.83±0.44	24.5±0.265
IIIa	Herbal tablet	10	30.67±0.67	48.67±0.88
IIIb		20	17±0.57	31.83±0.92
IIIc		30	8.33±0.67	17.66±0.43

The evaluation of anthelmintic activity on round worm revealed that the activity was dose dependent and inversely proportional to paralysis and death time of worms. From the graph (Figure 3 and 4), it is clearly seen that the herbal tablet shows significant activity as compared to reference control. The herbal tablet at dose 10mg/ml and 20 mg/ml shows significant activity at significant level of **p<0.001. The herbal tablet at 30 mg/ml dose has significant activity at significant level *p<0.01 in paralysis of round worm while it shows **p<0.001 significant activity in death time as compared to reference. The possible mechanism of the anthelmintic activity of polyherbal tablet cannot be explained on the basis of our present results. However, it may be due to its effect on inhibition of glucose uptake in the parasites and depletion of its glycogen synthesis. It may also have activated nicotinic cholinergic receptor in the worms resulting in either persistent depolarization or hyperpolarization ^[12].

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Received on 09.10.2015 Modified on 16.10.2015

Res. J. Pharmacology & P’dynamics. 7(4): Oct.-Dec., 2015; Page 181-183

DOI: 10.5958/2321-5836.2015.00036.1