49. An In Vitro evaluation of the Anthelmintic activity of Unripe Fruits extract of Carissa carandas Linn

Chanchal Kumar Mishra1*, D. Sasmal2 and B. Shrivastava3
  1. Department of Pharmacology, Arya College of Pharmacy, Kukas, Jaipur
  2. Department of Pharmaceutical Sciences, Birla Institute of Technology, Mesra, Ranchi
  3. School of Pharmaceutical Sciences, Jaipur National University, Jaipur, Rajasthan
Corresponding Author: Chanchal Kumar Mishra, Asst. Professor, Department of Pharmacology, Arya College of Pharmacy, Kukas, Jaipur, India. Email- [email protected]
Received:21 November 2012 Accepted: 06 December 2012
Citation: Chanchal Kumar Mishra, D. Sasmal and B. Shrivastava “An In Vitro evaluation of the Anthelmintic activity of Unripe Fruits extract of Carissa carandas Linn” Int. J. Drug Dev. & Res., October-December 2012, 4(4):393-397. doi: doi number
Copyright: © 2012 IJDDR, Chanchal Kumar Mishra et al. This is an open access paper distributed under the copyright agreement with Serials Publication, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Carissa carandas Linn is popularly known as ‘karunda’ in India belongs to family Apocynaceae. C. carandas Linn is a climbing shrub, usually growing to 10 or 15 feet (3-5 m) high. In folklore medicine, the unripe fruits of Carissa carandas Linn are Antidiarrhoeal, thermogenic, aphrodisiac, appetizer and anthelmintic activity, this is because of availability of specific and important phytoconstituents. The objective of the present work was to evaluate the in-vitro anthelmintic potency of the petroleum ether (60-80), chloroform and ethanolic unripe fruits extract of Carissa carandas Linn using Indian earthworms (Pheretima posthuma). The various concentrations (50, 100, and 150 mg/ml) of the different solvent extract were tested in in-vitro for anthelmintic potency by determination of time of paralysis and time of death of worm. Piperazine citrate (15mg/ml) used as standard drugs. The result of present study indicates that the unripe fruits extract of Carissa carandas Linn Potentiate to paralyze earthworm and also caused its death after some time. The shortest time of paralysis was observed at higher dose (150 mg/ml) of ethanolic extract (EECC), chloroform extract (CECC) and pet. ether extract (PEECC) i.e., found to 56.35 min, 40 min and 22.35 min respectively. The result showed that EECC took less time to cause paralysis of the earthworm than that of other unripe fruits extract of Carissa carandas Linn due to the availability of some important phytoconstituents. Thus the present studies demonstrate that the unripe fruits extract of Carissa carandas Linn having an anthelmintic has been confirm on Pheretima posthuma.


Carissa carandas Linn, Paralysis, death of earthworm.


Human intestinal helminthesis is among the most common infectious diseases occurring throughout the developing world. These infections have been associated with low standard of sanitation and the worldwide prevalence lies between 500 million to one billion annually approximately. It has been well accepted that due to the limited availability and affordability of modern medicines most of the world’s population depends to a greater extent on traditional medical remedies. Further, the use of medicinal plant products for treatment of various acute and chronic diseases is gaining increasing importance around the globe.1-3 It has been well evidenced that the traditional medicines including plants and plantderived preparations hold a great promise as source of easily available effective antihelmintic agents to the people
Carissa carandas Linn. (F; Apocynaceae) Its botanical name was in recent years changed to Carissa congesta. Wight (syn. C. carandas Auct., formerly widely shown as Carissa carandas Linn.) A genus of about 32 species distributed mostly in the warmer parts of India, Sri lanka, Java, Malaysia, Myanmar and Pakistan. Of the 8 Indian species, 3 are of economic importance. Its Leaves are simple, opposite, oblong-oval or oblong-lanceolate, subacute at the base, obtuse at the apex, glabrous and thin. The flowers are regular and bisexual. The fruit cluster of 3 to 10 is oblong, broad-ovoid or round, ½ to 1 inch (2.5-7.5 cm) long, has fairly thin but tough skin, purplish- red turning dark- purple or nearly black and shiny when ripe. 4-8 The sour unripe fruit is reputed for its aphrodisiac, appetizer, antipyretic, antimicrobial, astringent properties and is used in the treatment of diarrhoea and intermittent fever. The ripe fruit is used to treat mouth ulcer, sore throat and skin disorders. The root is used in healing stomach disorders and as an antihelmintic.[9-11]
There is no report on pharmacologically evaluated anthelmintic activity on unripe fruits extract of Carissa carandas till date.[12] Therefore, in this present study we have evaluated the antihelmintic activity of Pet. ether, chloroform and ethanolic extract of unripe fruits extract of Carissa carandas on Pheretima postuma.


Plant Materials and Authentication-

The unripe fruits and leaves were collected in May to July from Garhawa; Daltanganj and Daily Market, Ranchi (Jharkhand) and Ajmeri Gate Market, Jaipur (Rajasthan) and plant were identified by Dr. S. Jha, Professor, Department of Pharmaceutical Sciences, BIT Mesra, Ranchi. A Set of voucher Specimen was made for whole part of the plant. (Herbarium sheet No.- H-397/CM/D-2007)

Preparation of Extract-

The dried unripe fruit extract of Carissa carandas were powdered, defatted with petroleum ether and subjected to successive solvent extraction with chloroform, ethanol in Soxhlet extractor for 48 hours. All the extracts were further evaporated by rotary vapour Apparatus and the extract was collected. The preliminary phytochemical investigation was carried out for Ethanolic extract (EECC), Chloroform extract (CECC) and Pet. ether extract (PEECC) of unripe fruits of Carissa carandas. Further, the percentage of yield of EECC, CECC and PEECC were found to be 12.20, 8.23 and 10.12 respectively.

Administration of Extract:

The suspension of all extract of Carissa carandas with different concentration (50, 100 and 150 mg/ml) were prepared by using 0.2% v/v of Tween 20 as a suspending agent and final volume was made to 10 ml for respective concentration of unripe fruit extract of Carissa carandas Linn., Piperazine citrate (15mg/ml) was used as standard.[13-15]
Animals: Indian adult earthworms (Pheretima posthuma) were collected from water logged areas and were identified by Dr. N. K. Lohia, Department of Zoology, University of Rajasthan, Jaipur, (Rajasthan).

Anthelmintic activity against Pheretima posthuma-

Plant extract were investigated for their Anthelmintic activity against Pheretima posthuma. Various concentrations (10-150 mg/ml) of extract were tested in the bioassay, which involved determination of time of paralysis and time of death of the worms. Piperazine citrate (PC) is included as standard reference and distilled water as control.[16-17]
Eleven groups of approximately equal size earthworms consisting of six earthworms individually in each group were released into 10 ml of desired concentration of drug. The animals were divided into 11 group containing six earthworms of each different concentration of extracts and standard drug solutions were poured in different Petri-dishes. Observations were made for the time taken for paralysis (Paralysis was said to occur when earthworm did not revive in normal saline) and death (Time for death of worms was recorded after ascertaining that worms neither moved when shaken vigorously nor when dipped in warm water 50 ºC), followed with their body colors fading away. For evaluation of anthelmintic activity of unripe fruit extract of Carissa carandas plant, group I was control, group II received standard drug (Piperazine citrate), group III to XI received pet. ether (60-80), chloroform and ethanolic unripe fruit extract of Carissa carandas plant respectively. All the results were expressed as Mean ±S.D. of six animals in each group.[18-19]
Statistical Analysis: The data were analyzed with GraphPad Prism 4 (San Diego, CA). Statistical analysis of data was done by One-way ANOVA, followed by Newman Keuls test. Data are expressed as Mean ± Standard error of mean (S.E.M.). A level of P<0.05 was accepted as statistically significant.


Anthelmintics are drugs that may act locally to expel worms from the GIT or systemically to eradicate adult helminthes or development forms that invade organs and tissues. The pet. ether (60-80), chloroform and ethanolic unripe fruit of Carissa carandas plant were found to show anthelmintic activity when compared to standard drug. EECC plant of concentration (50, 100, and 150 mg/ml) showed paralysis at 58.00, 28.00 and 22.35 min and death at 121.56, 112.00 and 91.31 min while CECC plant of concentration (50, 100, and 150 mg/ml) paralysis at 87.32, 62.00 and 40.00 and death at 165.00, 140.00 and 119.66 min and PEECC showed paralysis at 98.22, 71.00 and 56.35 min and death at 221.56, 182.00 and 148.31 min. The standard drug, Piperazine citrate showed paralysis at 21.26 min and death after 72.33 min at 15mg/ml concentration. From the above result, it is clear that ethanolic, Chloroform and pet. ether (60-80) unripe fruit extract of Carissa carandas plant have significant anthelmintic activity in dose dependent manner when compared with standard anthelmintic drug.


From the results it concludes that, unripe fruit extracts of Carissa carandas (ethanolic, chloroform and pet. ether) demonstrate to possess dose dependant anthelmintic activity when compared to piperazine citrate. Data in the figure-1 and the present scientific work showed that EECC plant took the less time to cause paralysis of the earthworm than that of other solvent extracts thus it conclude that EECC possess potent anthelmintic activity. Preliminary phytochemical screening of the extracts revealed that the presence of terpenoids, flavonoids, alkaloids, tannins, saponins and steroids. It has been well established that PC by increasing chloride ion conductance of worm muscle membrane produces hyperpolarization and reduced excitability that leads to muscle relaxation and flaccid paralysis thus, our drug may have the similar profile of mechanism of action. Further, it has been reported that tannins which are polyphenolic compounds produce antihelmintic activity by binding to glycoprotein on the cuticle of the parasite and thus leads to death of the worm.20-21


The pet. ether, chloroform and ethanolic extract of unripe fruits of Carissa carandas showed anthelmintic activity on Pheretima postuma. Therefore, standardization of each extracts and isolation of phytoconstituents in each extracts for antihelmintic activity is required in the future.

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