Helmintholytic Activity of the Methanolic and Aqueous Extract of Seeds of Cleome viscose

 

Matsyagiri Lenkalapally*, Sumathi Cheruku, Shravya Koppula, Sindhuja Sandhi, Narayana Mandapu, Gowri Shankar N.L

Department of Pharmacognosy and Pharmacology, Swami Vivekananda Institute of Pharmaceutical Sciences, Vangapally, Yadagirigutta, Nalgonda-508286, Andhra Pradesh, India.

 

ABSTRACT:

The aim of the present study was to evaluate helmintholytic activity of crude methanolic and aqueous extract of seeds of Cleome viscosa using Pheretima posthuma as test worm. Albendazole was included as standard reverence and distilled water as control. The results of present study indicated that the crude methanolic extract significantly paralysis and also caused death of worm in short time, as compared to aqueous extract. The both extract showed greater P and D time than the standard reference albendazole. Various concentrations of (25, 50, 100 mg/ml) of methanolic and aqueous extracts were tested in the bioassay, which involved the determination of time of paralysis (P) and time of death (D) of the worms. Methanolic extract shows less time (2.88 min for paralysis and 8.64 min for death of worm with 100 mg/ml of methanolic extract of Cleome vicosa) for paralysis and death of the worms as compared to the aqueous extract in the study. In conclusion, the traditional use of seeds of the plant Cleome viscosa as an helmintholytic activity have been confirmed and further studies are suggested to isolate the active principles responsible for activity. These observations showed the helmintholytic activity of Cleome viscosa seeds of methanolic and aqueous extract in paralysis and death of Pheretima posthuma.

 

KEYWORDS: Albendazole, Cleome viscosa, Helmintholytic, Pheretima posthuma

INTRODUCTION:

Cleome contain about 427 species available as tropical and sub tropical of the world, among that seven species had medicinal significance are Cleome aspera, Cleome chelidonii, Cleome folina, Cleome gynandra, Cleome monophylla, Cleome tenella. Among these species, Cleome viscosa is a traditional folk herb found in throughout in india belongs to the family cleomaceae1. It is commonly known as sticky viscome in English, and also called as dog mustard in India, kukka vomita, kukkoti in telugu. Cleome viscosa is an annual herb, common weed in waste places in sand field, around villages and rocky crevices with 30-80 cm in height. Flowers appear as yellow color, in axillary or terminal racemes, flowers and fruits available throughout the year1,2.

 

Description of plant:

Cleome viscosa herb is erect, profusely branched herb; stems are in viscid, foetid, leaves 3-5 foliate, leaflets obovate, or ecliptic, ciliate, acute, flowers of yellow having 4 sepals, oblong, 4 petals oblong or obovate and stamens 12-18. Its ovary is glandular, unilocular ovules numerous parietal5. Capsules are terate, glandular hairy, dehiscing from above3. The isolates from the various parts of this plant has been used as a folk remedy since ancient times. Four verities of this plant parts are available, they are seed, root, bark and leaves4.

 

Causative Agents:

Antihelminthitic drugs are used in the treatment of helminthiasis (vomiting of worms)

The different types of worms include
i) Round worms: They are of two types, Adult Intestinal Nematodes and Larval Tissue Nematodes.
ii) Tape worms: They are found in the intestine or larva in the tissues.
iii) Flukes: These are hermaphrodites with an exception of blood flukes. They are found in blood vessels, the intestine, biliary tract, lungs.

 

Symptoms:
Symptoms of Helminthiasis are as follows
1)  Abdominal pain
2)  Diarrhea
3)  Fever
4)  Fatigue
5)  Enlarged liver
6)  Gastrointestinal inflammation
7)  Eosinophilia
8)  Dehydration.

 

Mode of Transmission:                                                                    
The disease is transmitted through
1) Fecal - oral route for ascaris, trichuris and hookworm
2) Skin penetration for hookworms

 

Prevention and Control:

Following measures can be taken for prevention and control of the disease

·        Personal hygiene

·        Environmental sanitation

·        Clean food and drinking water.

·        Use of slippers or shoes

·        The drugs used are Albendazole, Mebendazole and Levamizole Helminthiasis5, 6.

 

Characteristics of Cleome viscosa:

Table.1.Characteristics Cleome viscosa plant

Sl. no

Characteristics

Cleome viscosa

1

Habit

Glandular pubescent annual herbs, 30-80 cm tall

2

Leaves

3-5 foliolate, petioled, leaflets elliptic oblong, glabrous above, pubescent beneath, and sessile or sub sessile.

3

Flowers

1-1.5 cm across, solitary, axillary, in terminal lax racemes.

4

Capsules

4-8 cm long, cylindric, viscid, and minutely beaked.

5

Seeds

Numerous, dark brown, reniform, 1.2 mm long

6

Flowering and Fruiting

.Through out the year

 

 

Figure.1. Diagram of Cleome viscosa plant.

Traditional and other medicinal uses:

·        In traditional systems of medicine the plant is reported as an antihelmintic, antiseptic, carminative, anticorbutic, sudoprific, febrifuge and cardiac stimulant.

·        The rural people use the fresh juice of crushed seeds of this plant for infantile convulsions and mental disorders7.

·        The pungent seed can be pickled or used as a mustard substitute in curries and the oil of seed used for cooking.

·        The pharmacological studies have shown that Cleome viscosa possesses various notable biological activities such as anthelmintic, antimicrobial, analgesic, anti-inflammatory, immunomodulatory, antipyretic, Antidiarrheal, psychopharmacological, and hepatoprotective activities8.

·        The leaves are prescribed in external applicable like inflammation of the middle ear, for healing of wounds and ulcers, dried and powdered seeds are valued as analgesic, anthelmintic.

·        A hot infusion of the root is used as restore consciousness, treatment of scurvy and rheumatic problems9.

·        The whole plant is also considered as a remedy for liver diseases, cardiac disorders, bronchitis, flatulence, colic, dyspepsia, constipation and cough10.

 

Phytochemistry:

Cleome viscosa is a rich source of elements like nitrogen, potassium, phosphorous, calcium, iron manganese, zinc, sodium, chloride, magnesium, copper, boron, silicon and nutritional vitamin like vitamin C, source of flavonoids, anthraquinone glycosides, phenolic compounds, tannins, steroids, glycosides, fatty acids (linoleic acid, palmitic acid, stearic acid, oleic acid ) beta carotene, saponins, coumarin lignanes2,11. Phosphorous is tied to calcium in bone structure and play a significant role in CNS function, many enzymes contain as a base phospoprotein, phospholipids are involved in nerve conduction, and phosphate is a primary ion in extracellular and intracellular fluid. Potassium plays a major role in the treatment of diabetes as it effect on the secretion of insulin. Potassium deficiency may cause symptoms of fatigue, weakness, mental depression, abnormal heartbeat, dry skin, low blood pressure and muscle cramps. Calcium also plays a major role in CNS function, is major factor in neurotransmission and important in nerve impulses conduction. Manganese deficiency causes skeletal abnormalities, retard bone growth, change in hair color, and abnormal changes in lipid and carbohydrate metabolism. Zinc deficiency may be associated with mental lethargy, emotional disorders and irritability. Copper is essential for the synthesis of dopamine, copper deficiency may causes hypertension, antibiotic sensitivity, hyperactivity, hyperglycemia, manic disorders, insomnia, allergies and osteoporosis. Boron enhances brain function, promotes alertness, control cell growth and regulate the body uses the calcium, phosphorous and magnesium. Sodium maintains the acid alkali balance of the body12, 13,.

 

MATERIALS AND METHODS:

Collection of plant:

·        The seeds of Cleome viscosa were collected and authentified by senior research officer in pharmacognosy department.

 

Preparation of plant extract:

·        The plant material (seeds) were collected, dried, and powdered as coarse particle.

·        Then defatting the powder of seeds using petroleum ether (40-60 0 C)

·        It was dried and extracted with methanol14.

·        The liquid extract was evaporated in vacuum to yield 34.5 %.

·        The methanol extract was carried out by soxhlation for 72 hrs and aqueous extraction was carried out by maceration method for 7 days15.

·        The solvents were removed under reduced pressure to yield 9.0 % and 7.5 % for methanol and aqueous extracts respectively16, 17.

 

Collection of worm and authentication:

·        Indian earthworm Pheretima posthuma (annelida) were collected from the water logged areas of soil and authentified at PG department of zoology.

 

Helmintholytic activity:

·        The activity was performed on Indian earth worms due to its anatomical and physiological resemblance with the intestinal round worm parasite of human intestine.

·        Indian adult earth worm (Pheretima posthuma) of 5-8cm in length and 0.2-0.3cm in width were used18.

·        Eight groups of approximately equal sized earth worms each containing six earth worms were selected19.

·        All the earth worms were washed in normal saline solution before they were used.

·        The control group was treated with distilled water.

·        The earth worms were placed in a standard and extracts and time of paralysis (P) and time of death (D) were calculated20.

·        The time of paralysis was noted when no movement of any sort could be observed expect when the worms were shaken vigorously.

·        The time of death were recorded after ascertaining that the worms neither moved when shaken vigorously nor when dipped in warm water (500C) 21, 22.

 

RESULTS AND DISCUSSION:

·        Primary phytochemical screening of crude alcoholic extract detects some secondary plant metabolites like steroids, alkaloids, saponins and flavanoids.

·        Helmintholytic activity of the methanolic and aqueous extracts of seeds of Cleome viscosa (L) was shown in table 1.

·        From the experimental work standard drug albendazole at Pheretima posthuma are 9.36±0.05 and 14.02±0.57mins respectively23.

·        Albendazole involves inhibition of glucose uptake system leading decrease in energy reserves in the helminthes to cause paralysis so that they are expelled in the feaces of man and animals.

·        Both the extracts of methanolic and aqueous extracts of seeds of Cleome viscosa were found to possess interestingly good level of helmintholytic activity24.

·        Methanolic extracts of seeds of Cleome viscosa and its different fractions exhibited helmintholytic activity in dose-dependent manner giving shortest time of paralysis (2.88±0.19 min) and death (8.642±0.38 min) with 100 mg/ml concentration as compared to that of aqueous extracts25.

·        The seeds of Cleome viscosa (L) not only resulted paralysis but also caused death of worms especially at higher concentration (100 mg/ml) of methanolic extracts in shorter time as compared to reference drug Albendazole26. 

 

Table.2. Helmintholytic activity of the methanolic and aqueous extracts of seeds of Cleome viscosa

Test

Substance

Concentration

(mg/ml)

Time taken for paralysis (P) and death (D) of earth warms

P                              D

Albendazole

25

09.36±0.05           14.02±0.57

Methanol extract

25

50

100

06.11±0.25           20.28±0.57

04.94±0.28           15.46±0.57

02.88±0.19           08.64±0.38

Aqueous extract

25

50

100

20.00±0.84           88.22±3.18

16.28±0.90           62.45±2.14

12.40±0.63           22.62±1.73

Control

-

-                 -

All values represent Mean ± SEM; n ═ 6 in each group; P: Paralysis time, D: Death time; SEM – Standard error of mean; control worms were alive up to 24 hrs of the experiment; n= number of animals (earth worms).

 

CONCLUSION:

This study has revealed that this plant Cleome viscosa has many secondary metabolites (Phytoconstituents) and shows dose dependent helmintholytic activity and albendazole shows less paralysis and death time as compared to others. Various concentrations of (25, 50, 100 mg/ml) of methanolic and aqueous extracts were tested in the bioassay, which involved the determination of time of paralysis (P) and time of death (D) of the worms. Methanolic extract shows less time (2.88 min for paralysis and 8.64 min for death of worm with 100 mg/ml of methanolic extract of Cleome viscosa) for paralysis and death of the worms as compared to the aqueous extract in the study. Further, it would be interesting to isolate the phytoconstituents responsible for helmintholytic potential.

 The present research work was providing a basis for further detailed investigations in the direction of isolation of other active compounds.

 

 

ACKNOWLEDGEMENT:

The task of preparing this dissertation has been fascinating experience and it is really a moment of great pleasure for me to express my hearty gratitude to those who have helped me in successfully completion of this dissertation. I express my sincere thanks to Reddy’s Labs for providing gift sample of drug. We thankful to the management of Swami Vivekananda Institute of Pharmaceutical Sciences, Vangapally, Yadhagirigutta, Nalgonda, Andhra Pradesh, India, for providing all facilities during the study.

 

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Received on 07.05.2012

Modified on 18.05.2012

Accepted on 30.05.2012

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Research J. Pharmacology and Pharmacodynamics. 4(5): September –October, 2012, 259-262