In vitro anthelmintic and antioxidant potential of fruits of Momordica charantia: A comparative study

Saikat Sen; Raja Chakraborty; Bitul Borah; BK Dey; Bapi Ray Sarkar; BJ Sahariah

doi:10.4103/2349-5006.148813

ORIGINAL ARTICLE

Year : 2014 | Volume : 7 | Issue : 2 | Page : 113-117

In vitro anthelmintic and antioxidant potential of fruits of Momordica charantia: A comparative study

Saikat Sen, Raja Chakraborty, Bitul Borah, BK Dey, Bapi Ray Sarkar, BJ Sahariah
Institute of Pharmacy, Assam Down Town University, Panikhaiti, Guwahati, Assam, India

Date of Web Publication

7-Jan-2015

Correspondence Address:
Dr. Saikat Sen
Institute of Pharmacy, Assam Down Town University, Panikhaiti, Guwahati - 781 026, Assam
India

Source of Support: None, Conflict of Interest: None

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DOI: 10.4103/2349-5006.148813

Abstract

Background and Purpose: Momordica charantia L. (Cucurbitaceae) is used as both food and management of diverse human diseases science ancient time, including in the treatment of worm infection. This study was aimed to evaluate the comparative anthelmintic and anti-oxidant activity of juice and extracts of whole fruit, peels, and seed of M. charantia.
Materials and Methods: In vitro anthelmintic activity of methanol extract of whole fruit, fruit peels, seed and fresh juice whole fruit, peels of M. charantia was investigated against Indian adult earthworms (Eisenia foetida). Extracts and juices are also investigated for their anti-oxidant activity using 2,2-diphenyl-picrylhydrazyl radical (DPPH^• ) scavenging assay method.
Results: Extracts and juice showed potent anti-oxidant and anthelmintic activity. Methanol extract of fruit peel demonstrated strong DPPH^• scavenging activity (IC ₅₀ = 29.02 ± 0.86 μg/ml), juice of fruit peel also produced good radical scavenging effect (57.30 ± 0.99 μmol TE/ml). Methanol extract of fruit peel showed potent anthelmintic effect which is similar to standard albendazole. Whole fruit and seed extract also produced significant anthelmintic effect. Whole fruit juice and peel juice showed similar but moderate in vitro anthelmintic effect.
Conclusion: The wormicidal activity of extract and juice against earthworms suggests that they are effective against parasitic infections of humans. Results justify the use of fresh kerala juice by traditional medicinal practitioner to cure worm infection, and showed the potential to develop natural anthelmintic constitute from the fruit of the plant.

Keywords: Anti-oxidant, fruit, Momordica charantia, worm infection

How to cite this article:
Sen S, Chakraborty R, Borah B, Dey B K, Sarkar BR, Sahariah B J. In vitro anthelmintic and antioxidant potential of fruits of Momordica charantia: A comparative study. Indian J Health Sci Biomed Res 2014;7:113-7

How to cite this URL:
Sen S, Chakraborty R, Borah B, Dey B K, Sarkar BR, Sahariah B J. In vitro anthelmintic and antioxidant potential of fruits of Momordica charantia: A comparative study. Indian J Health Sci Biomed Res [serial online] 2014 [cited 2022 Oct 4];7:113-7. Available from: https://www.ijournalhs.org/text.asp?2014/7/2/113/148813

Introduction

Momordica charantia L. (family: Cucurbitaceae) is a tropical and subtropical vine distributed in Asia, Africa, South America, Caribbean countries and commonly known as Karela in India. The plant is cultivated for its bitter fruits, which widely used as a vegetable. Since ancient times, different parts of M. charantia are employed in traditional management of diverse human diseases. ^[1] Fruit of M. charantia is highly valued for its anti-diabetic, hepatoprotective, anti-gout, anti-rheumatism and anti-microbial properties. Leaf of the plant is used to cure leprosy, piles, jaundice, cough and chest pain in children. The seed of the plant has been regarded to reduce diabetes and fat. Shoots are used by some ethnic people to treat pneumonia and leukorrhagia while roots possess abortifacient activity. Fruit, leaf, seeds of the plant used as anthelmintic in different traditional medicinal systems. ^[2],[3],[4] Previous biological studies of M. charantia showed that the plant parts possess anti-oxidant, ^[5],[6] anti-diabetic, ^[7],[8] anti-obesity, ^[9] anti-viral, ^[10] anti-bacterial, ^[11] insecticidal, ^[12] anti-cancer, ^[13] anti-tumor, ^[14] anti-depressant and anxiolytic, ^[15] anti-fertility, ^[16] wound healing, ^[17] analgesic and anti-pyretic ^[18] activity. The in vitro and in vivo trial has shown that 3% aqueous extract of M. charantia whole fruit possessed anthelmintic activity against Ascaridia galli.^[19] Lyophilized M. charantia extracts were investigated for its anthelmintic activity and found that at 500 μg/ml extract shown potent activity against Caenorhabditis elegans. ^[20] Ethanol extract of leaf has investigated for in vitro anthelmintic effect against Ascaris suum. ^[21] Another study reported that plant extract has better anthelmintic effect against A. galli than piperazine hexahydrate. ^[22] M. charantia contains a number of potential pharmacologically active constituents such as charantine, goyaglycoside, mormodicoside, mormodicoside 3β, 25-dihydroxy-5β, 19-epoxycucurbita-6, (23E)-diene, momordicine-I, karavilagenin, karavilagenin C, karaviloside, karaviloside, kuguacin, kuguacin A, kuguacin B, kuguacin E, ^[4],[23] 3, 7, 23-trihydroxy-cucurbita-5,24-diene-19-al, ^[23] 3, 7, 25-trihydroxy-cucurbita-5, 23-diene-19-al, ^[23] 3,7-dihydroxy-25-methoxycucurbita-5, 23-diene-19-al. ^[23]

In tropical developing countries helminthic infections possess a major health hazard to the people. Even though gastrointestinal helminthic infections not responsible for significant morbidity and mortality but they may responsible for substantial effects such as malabsorbtion, diarrhea, anemia and other states of poor health. ^[24],[25] In spite of the presence of a large number of synthetic medicines people still do not have access to, or cannot afford it especially in remote areas. Drug resistance is another key problem to manage parasite infections. In this context, use of medicinal plants tender a foremost and accessible source of health care to people. ^{[24],[25],[26]}

Anthelmintic potential of different parts of M. charantia including fruits and seeds were described in traditional medicinal knowledge, and some current investigations have also reported the anthelmintic effect of M. charantia fruit. ^{[2],[3],[4],[20],[21],[22]} Peoples of India usually consumed juice of both fruit peels and seed, and also as a vegetable. However, no comparative study was available, keeping in view their widespread utilization, the present study was aimed to evaluate the comparative anthelmintic and anti-oxidant activity of juice and extracts of whole fruit, peels and seed of M. charantia against Indian adult earthworms (E. foetida).

Materials and Methods

Plant materials

Mature fruits of M. charantia L. were collected in December 2013 from the local market of Guwahati, Assam. Plant material was identified and authenticated by Dr. AA MaO, Senior Scientist, Botanical Survey of India, Shillong, India (No: BSI/ERC/2014/Plant identification/786).

Preparation of juice and extract of M. charantia fruit parts

Whole fruits were cut into small pieces, and air dried under shed. Dried fruit parts were pulverized to a fine powder using a mechanical grinder. The powder was extracted with methanol using Soxhlet apparatus. The extract was concentrated by distillation and solvent was evaporated to dryness on the water bath to get the methanol extract of whole fruit of M. charantia (MWF).

To obtain the methanolic extract of fruit pulp (MFP), seeds were removed from the fruit and then fruit pulps were dried under shed. The dried fruits pulps were pulverized to a fine powder and extraction was carried out using methanol in Soxhlet apparatus. The extract was concentrated, and solvent was evaporated to dryness.

A similar procedure was carried out by taking the seeds only to obtain methanolic extract of fruit seed (MFS).

The fresh whole fruits were collected and washed properly. The 50 g of fruit was grinded using a mechanical grinder and screened through fine sieve to collect the juice of whole fruit (JFW). Similar protocol was used after removing the seeds to obtain juice of fruit pulp without seed (JFP).

Preliminary phytochemical test

All extracts and juice were investigated to find the presence of different phytochemicals using the standard methods. ^[27],[28]

In vitro anti-oxidant activity

Extracts and juice were investigated for in vitro anti-oxidant effect using 2,2-diphenyl-picrylhydrazyl (DPPH) radical scavenging assay method. DPPH radical scavenging assay method described by Sen et al. ^[29] was used to find the anti-oxidant potential of extracts, while method explained by Jakob et al. ^[30]

Experimental animals and anthelmintic activity

Indian adult earthworms (E. foetida) were used for anthelmintic activity. Earthworms were authenticated and supplied by North East Green Tech Private Limited, Bamunimoidam Guwahati, Assam, India.

Indian adult earthworms were washed in normal saline to remove all fecal matter. Earthworms about 3-5 cm in length and 0.1-0.2 cm in width were taken for the study. Indian earthworms are divided into 10 groups each containing six earthworms in the following manner:

Group 1: Control (Normal Saline, 0.9% NaCl)
Group 2: Standard (Albendazole suspension) (40 mg/ml)
Groups 3 and 4: Methanol extract of whole fruit (MFW) (100 and 150 mg/ml)
Groups 5 and 6: MFP (100 and 150 mg/ml)
Groups 7 and 8: MFS (100 and 150 mg/ml)
Group 9: JFW (30 ml)
Group 10: JFP 30 ml.

Indian earthworms of each group were released into 30 ml of solution of respective standard and test drugs. Observations were made for paralyze time and death time for individual worms as per the procedure described by Sen et al. ^[31]

Statistical analysis

The results are expressed as mean ± standard error of the mean the statistical difference was tested by using one-way analysis of variance followed by Tukey tests. P <0.001 was used as a criterion for statistical significance.

Results and Discussion

Momordica charantia fruits and seeds are known for their wormicidal activity since ancient time in Indian societies. People consume fruit and seed as vegetable, and also takes juice of the fruit to remain healthy. Few studies have investigated their anti-oxidant activity and the potency of whole fruit, leaves against different helminth species. ^{[2],[3],[4],[20],[21],[22]} This study was undertaken to compare the activity of fruit, seed in a different form against Indian earthworm.

Among the three extracts, the highest yield was observed for MFP (21.94% w/w) followed by MFW (16.9% w/w), MFS (8.56% w/w). While yield of the JFP was found 18 ml/100 g fruit pulp and 16 ml/100 g whole fruit for JWF. Preliminary phytochemical test showed that chemical constituents such as glycoside, alkaloid, carbohydrate, saponin were commonly present in extract and juice.

Among the extracts, MFP showed highest DPPH^• scavenging activity (IC ₅₀ = 29.02 ± 0.86 μg/ml), followed by MFW, MFS. JFP showed better radical scavenging effect (57.30 ± 0.99 μmol TE/ml) than JFW (45.41 ± 0.42 μmol TE/ml) [Table 1]. DPPH is a stable free radical, on reaction with anti-oxidant the electron of DPPH^• is paired off, which results in decolorization of DPPH solution. ^[32],[33] The strong scavenging capacity of the extracts and juice indicated this effect in a part might be responsible for biological activity.

Table 1: Antioxidant activity of extracts and juice of whole fruits, peels and seed of M. charantia

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Methanolic extract of fruit pulp at 150 mg/ml showed almost similar effect on paralysis (8.50 min) like that of albendazole (8175 min), while higher concentration of MFP found more effective in terms of death time (14.50 min) than standard drug (16.33 min). MFS and MFW also demonstrated potent anthelmintic effect. Among the juice, JFP showed quick paralyzis time (22 min) while JFW produce quick death to the worms (14.5 min). Both the juices showed almost similar activity [Table 2]. Resistance toward the available synthetic medicine possesses a serious problem worldwide. Plant-derived drugs are considered a potent source of new anti-microbial agents. ^[31] Due to the easy availability, anatomical resemblances with intestinal worms and their reaction to anthelmintics, ^[31] Indian adult earthworms (E. foetida) have been used for this comparative evaluation.

Table 2: Comparative anthelmintic activity of juice and extracts of whole fruits, peels and seed of M. charantia

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Preliminary phytochemical analysis and previous investigations confirmed that fruit and seed contain tannins, glycosides along with other chemical constituents. These phytochemicals can attach with free proteins in the gastrointestinal tract or glycoprotein on the parasite cuticle and cause deaths. Anti-oxidant activities of extract and juice could have contributed, at least partly, to the therapeutic benefits of M. charantia fruit against worm infection. The wormicidal activity of extract and juice against earthworms suggests that they are effective against parasitic infections of humans. Results showed that the extract from fruit pulp showed better activity that is similar to that of standard drug. Study confirmed that fresh JFP exhibited potent activity against Indian earthworm, which justify the use of Kerala juice by traditional medicinal practitioner to cure worm infection.

Conclusion

In summary, this study highlight that extract and juice of M. charantia whole fruit, peels, seed have potent anti-oxidant and anthelmintic effect. Fruit peel extract exhibited very good anthelmintic effect, thus results give the experimental basis to understand the use of juice of the fruit in traditional medicine, as an anthelmintic agent. In view of the potential use of M. charantia fruit in the traditional medicine, its therapeutic benefits and bioactive compounds warrant further investigation.

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