An In-vitro Evaluation on Anthelmintic activity of different extracts of Hemigraphis colorata leaves

 

Sabin Scariya, Dijo Davis, Jahanara Hameed, Anjusha M. K, Merin Babu*

Department of Pharmacology, Amrita School of Pharmacy, Amrita Vishwa Vidyapeetham,  Kochi – 682041, Kerala, India

 

ABSTRACT:

Objective: Worm infection is a serious problem affecting millions of people all over the world. Traditional chemical drugs are not fully effective due to the evolving drug resistant worm strains, so exploring new remedies derived from natural products is a good way to fight these infections. Hence, the present study was undertaken to determine the anthelmintic activity of leaves of Hemigraphis colorata. Methodology: The study was carried out on Indian earthworms (Pheretima posthuma). The effects of different concentrations of fresh juice extract, ethanolic extract and aqueous extract of the leaves were compared with standard drug Albendazole (20 mg/ml). Distilled water was taken as the control. The time for paralysis (reduction in motion and movement of worm) and death of the worms (no movement) were noted. Result: All the extracts (fresh juice, ethanolic and aqueous) of leaves showed anthelmintic property. The fresh juice extract showed the best result compared to the other two extracts. The fresh juice extract showed the paralysis and death time at 12 min and 21 min. respectively. In case of Albendazole group, paralysis and death time was at 6 min and 10 min. respectively. Conclusion: The results obtained confirm the anthelmintic property of the plant. However, further studies are required to elucidate the mechanism responsible for the beneficial effect.

 

 

INTRODUCTION:

Even though parasitic worm infections are a serious issue, the studies in the field of anthelmintic drugs are very less [1]. Anthelmintics are those agents that expel parasitic worms from body either by stunning or by killing them [2]. Anthelmintics obtained from natural sources are a key to the treatment of these parasitic worm infections. Lately, significant consideration has been give to utilize herbal medicine for treatment of these types of various diseases [3]. There are a huge number of natural herbs which shows potential anthelmintic activity. Hemigraphis colorata is an example for one of such species.

 

Hemigraphis colorata of the Acanthaceae family is a low-creeping perennial herb. It reaches an average height of about 15 to 30 cm. The plant has numerous synonyms like aluminium plant, metal leaf, cemetery plant, red flame ivy, waffle plant, java ivy, etc. This is an excellent garden and showcase plant. The plant is known as 'murikootti' and 'murian pacha' in malayalam. The plant got its name because of its high wound healing property. The leaves show purple colour. They are ovate and opposite with an average size about 2-8cm in length and 4-6cm wide.

 

The plant contains constituents like saponins, phenols, terpinoids, flavanoids, carboxylic acids, coumarins, carbohydrates, proteins, xanthoproteins, tannins, steroids, sterols and alkaloids. The leaf shows presence of constituents like of polyphenols, flavanoids, tannins together with low sodium and high potassium level [4].

 

Though it is a showcase plant, it has several pharmacological activities. The plant was used as a natural herb in ancient medicine. The plant was found to have uses like anti-bacterial activity, anti-diabetic activity, wound healing property [5], anti-oxidant activity, etc.

 

Earthworms (Pheretima posthema) commonly known as the farmers friend are used in this anthelmintic study due to the easy access to them. Earthworms can easily be spotted and caught from fields and other wet areas. The study on the anthelmintic property of the leaf extract of Hemigraphis colorata was carried out using the Indian earthworms. The study was carried out on earthworms due to their anatomical resemblance with intestinal round worms [6]. The rate of death of earthworms was noted during the study.

 

MATERIALS AND METHODS:

Collection of Plant Material:

The fresh leaves of Hemigraphis colorata were collected from regions of Pala, Kottayam district. The plant was authenticated and identified by Dr. Giby Kuriakose. The fresh leaves required for the preparation of juice extract of the leaves were collected at the time of study. The leaves to be extracted were collected few days before the study and were cleaned and kept in shade for drying.

 

 

Figure 1: Hemigraphis colorata

 

Preparation of Extract:

Preparation of Fresh juice extract:

The leaves which were to be extracted were collected at the day of study and was washed and cleaned. The clean leaves were crushed using mortar and pestle and the fresh juice was strained out. The obtained juice was filtered using whatsmann filter paper and was collected. The obtained extracts were dried by evaporating.

 

Preparation of Aqueous extract:

The earlier dried leaves were powered and were placed in a stoppered iodine flask. Water was added to the iodine flask and was shaken well. This was kept for seven days for maceration. The flask was shaken at regular intervals. After seven days the mixture was filtered and the extract was obtained. The obtained extract was dried by heating.

 

Preparation of Ethanolic extract:

The earlier collected and dried leaves were powdered in a mixer grinder and was weighed and packed and subjected to hot continuous soxhlet extraction at 40⁰ C. The product obtained was concentrated first by distillation and then the extract was dried by evaporating it in a water bath at 40-50°C.

 

Standard drug:

The standard drug Albendazole was purchased from a local pharmacy store.

 

Collection of test organism:

The Indian earthworms were collected from wet and moist local regions. The collected worms had an average size around 5-7cm. These worms were cleaned using normal distilled water and all the soil and dirt particles were removed [7].

 

Anthelmintic property study:

The test was carried out according to the standard procedures of Ajeyoba et al with slight modifications; the organisms were divided into different groups: Control, standard and test. All groups contained five worms each. Control contained normal saline and distilled water. The standard drug used was Albendazole. The standard was also prepared in different concentration (20mg/ml,40mg/ml and 80mg/ml). The test contained three sub groups of aqueous extract, ethanolic extract and fresh juice extract. Each sub group had three different concentrations (20mg/ml,40mg/ml and 80mg/ml). The worms were added to the prepared solutions and time was noted. The time taken for the paralysis and death of the worms in each group was observed. The time when the motility of worms were reduced to a great extent; that they moved only on shaking was observed as the time of paralysis. The time of death was noted when the worms showed no movements during vigorous shaking. The death was confirmed by putting a worm each from each group in slight warm water and observed their movements, if the worms were immobile then the death was confirmed [8,9,10,11].

 

RESULT AND DISCUSSION:

The observations prove positive anthelmintic activity for the leaves of Hemigraphis colorata. The results showed a progressive increase in the rate of paralysis and death with increase in concentration of the extracts. The higher concentration of fresh juice extracts showed a comparative activity to the standard Albendazole. The activities of ethanolic and aqueous extracts were less than that of fresh juice extracts. The results are shown in Table 1. From the obtained results it is clear that the standard Albendazole (80mg/ml) showed the effect at 10 minutes and the 80mg/ml concentration of fresh juice extract showed the effect at 21minutes where as the higher concentrations of aqueous and ethanolic extracts showed the effect at 28 and 43 minutes respectively.

 

Table 1: Anthelmintic activity of Hemigraphis colorata

 

The significant anthelmintic activity of the extracts of leaves of Hemigraphis colorata may be due to the presence of tannins, saponins or flavanoids. Polyphenolic compounds are also shown to cause anthelmintic activity [12, 13].

 

In short the anthelmintic property of the plant may be because of any of the above said constituents. Further detailed studies have to be conducted with the isolated constituents of the plant to finalise the mechanism of death of the worms.

 

 

Figure 2: Different concentrations of extracts

 

CONCLUSION:

The anthelmintic effect of the different extracts of the leaves of Hemigraphis colorata was observed. The observations were given in Table1 and Figure 2. The extracts showed comparative results with the standard Albendazole. The fresh juice extract gave the best results comparing to the other extracts. The aqueous extract showed the next best result. The better results of the fresh and aqueous extracts may be because of the presence of more amounts of phyto constituents due to higher elution. The dose dependent activity of the leaf extracts proves the anthelmintic property of the plant so further studies has to be conducted in order to better understand the proper mechanism of action of the extracts.

 

ACKNOWLEDGEMENT:

The authors would like to show their humble gratitude and thankfulness towards the principal and management of Amrita School of Pharmacy for providing all necessary arrangements and requirements for carrying out the study.

 

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Accepted on 20.05.2019          © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(9):4394-4396.