Ijpcr.netC. Velmurugan,et al / Int. J. of Pharmacology and Clin. Res. Vol-1(1) 2016 [1-7]
International Journal of Pharmacology and
Clinical Research (IJPCR)
IJPCR Volume 1 Issue 1 Jan-Mar - 2016
Research article Pharmacology research
Anthelmintic activity of leaves of different extracts of Gossypium
C.Velmurugan*1, Sujith Thomas2, Anurag Bhargava3, SK Shajahan1
1Department of Pharmacology, Sri Krishna Chaithanya College of Pharmacy, Madanapalle-517325,
Andhra Pradesh, India.
2Department of Pharmacology, Moulana College of Pharmacy, Kerala-679321, India.
3Department of Pharmacognosy, CH. Devi lal College of Pharmacy, Bhagwangarh, jagadhri, Haryana-
*Corresponding author: C.Velmurugan
E-mail id: firstname.lastname@example.org
The N-hexane, ethyl ether and ethanol extracts of leaves of Gossypium herbaceum L was investigated for anthelmintic
activity using earthworms (Pheretimaposthuma). Various concentrations (10,20,40,60,80 &100 mg/ml) of plant extracts were
tested in the bioassay. Albendazole (10 mg/ml) was used as reference standard drug whereas 1% v/v tween 80 as control.
Determination of paralysis time and death time of the worms were recorded. The ethyl ether and ethanol extracts exhibited
significant anthelmintic activity at highest concentration of 60, 80 & 100 mg/ml compared to standard drug. The result shows
that ethyl ether extract possesses potent vermicidal activity and found to be effective as an anthelmintic compared to
Keywords: Anthelmintic, Gossypium herbaceum, Albendazole, Earthworm and Pheretimaposthuma.
Helminthes infection, the source of infection being very In the recent years, the importance of Herbal drugs in common due to poor sanitation, poor family hygiene, Medicine has tremendously increased because of their malnutrition, and crowded living conditions (Basu& fewer side effects. Consequently, the demand for the Sharma 2005). Although the majority of infections due to herbal formulation is increasing day by day. The helminths are generally restricted to tropical regions and phytochemical constituents and their standardization are cause enormous hazard to health and contribute to the accelerated with the development of instrumental analysis prevalence of undernourishment, anaemia, eosinophilia and this field becomes important and new for and pneumonia (Bundy1994). Parasitic diseases cause ruthless morbidity affecting principally population in Helminth infections are among the most widespread endemic areas (Tagbota&Townson2001). The gastro- infections in humans, distressing a huge population of the intestinal helminthes becomes resistant to currently world. As the half of world suffering from bacterial and available anthelmintic drugs therefore there is a foremost problem in treatment of helminthes diseases and treatment C. Velmurugan,et al / Int. J. of Pharmacology and Clin. Res. Vol-1(1) 2015 [1-7]
of helminthic infections becomes a major concern today district, Andhra Pradesh. The leaves were shade-dried and (Sondhi&Shahu1994). Hence there is an increasing powdered and stored in airtight containers further use. demand towards natural anthelmintics. The disease is highly prevalent particularly in third world countries (Dhar et al 1982) due to poor management The plant under investigation was identified with the help practices. Chemical control of helminthes coupled with of taxonomists, Prof. Dr. T Damodharam, M.Sc, M.Phil, improved management has been the important worm Ph.D, Assistant Professor, Sri Venkateswara University, control strategy throughout the world. However, increasing problems of development of resistance in helminthes (Geert et al 1995, Coles 1997) against anthel- Preparation of extracts
mintics have led to the proposal of screening medicinal The collected leaves were shade dried completely. The plants for their anthelmintic activity. The plants are dried leaf was then coarsely powdered and was sieved known to provide a rich source of botanical (sieve # 60) to get uniform powdered. The 1kg powder anthelmintics(Satyavati et al 1976, Lewis 1977)A number material extracted with solvent N-hexane, ethyl ether of medicinal plants have been used to treat parasitic and80% ethanol in a Soxhlet apparatus. Final compound infections in man and animals (Nadkarnia 1954, Akhtar et was concentrated by vacuum drying. The traces of the solvents were removed by keeping the dried extracts in to The Survey of literature reveals that the medicinal plant of Gossypium herbaceum Linn. Malvaceae is used in Ayurveda to treat various diseases. In Ayurveda the Preliminary phytochemical screening
properties of Karpasa are Katu (Pungent), Kashaya The fractions of leaves of Gossypium herbaceum was (Astringent) in rasa; Laghu (Light), Tikshna (Penetrating) screened for the presence of various phytoconstituents guna; Ushna (Hot) virya and Katu (Pungent) vipaka. It is like alkaloids, flavonoids, saponin, tannin and glycosides etc. (Kokate 1986) (Dysmenorrhoea) and Prasutipashchatavikara (Purpueral disorders) (Chunekar et al 1982, Sharma et al 2001). collection
2007)and root bark is anticancerous (Jain 1991). The appropriate time for their collection was found early Gossypium herbaceum contains a Gossypol. Gossypol is a in the morning in the summer, and noontime during the male contraceptive. It also assists menstrual flow and winter. Freshly collected alive worms were stored in the effectively inhibits egg implantation (Choudhry et al plastic bags, filled with suitable quantity of wet compost 1980, Krishna Reddy et al 1984). Gossypol and its derivatives have been shown to have significant antimicrobial activity as well as wound healing effect Evaluation of anthelmintic activity
(Reddy et al 1981).In traditional system of medicine the An Indian adult earth worm 4 -5 cm in length and 0.1 - tribals use various indigenous plants for the treatment of 0.2 cm in width were used for the in vitro anthelmintic bio anthelmintic. One such plant drug used by tribals many assay. Because of easy availability, earthworms have parts of Andhra Pradesh is leaves juice of Gossypium been used widely for the initial evaluation of anthelmintic herbaceum commonly called cotton plant is claimed by compounds in vitro(Sollmann 1918). The anthelmintic folklore for anthelmintic. With this view, the Gossypium assay was carried out as per the method. (Ajaiyeoba et al herbaceum is studied for its anthelmintic activity. 2001). The assay was performed in vitro using adult earthworm (Pheretimaposthuma) owing to its anatomical and physiological resemblance with the intestinal MATERIALS AND METHODS
roundworm parasites of human beings for preliminary Plant material Procurement of leaves
evaluation anthelmintic activity (Vigar 1984, Dash et al The leaves used for the investigation were collected from 2002, Shivakumar et al 2003) a medicinal garden at a place called sri Krishna chaithanya college of pharmacy, madanapalle in chittoor C. Velmurugan,et al / Int. J. of Pharmacology and Clin. Res. Vol-1(1) 2015 [1-7]
The earthworms were divided into different groups containing six worms in each group. Preliminary phytochemical screening
Control group received 1% v/v of tween80. The leaves of The preliminary phytochemical analysis of fractions of N-hexane, ethyl ether (EEEGH) and ethanol extracts Gossypium herbaceum shows presence of steroids, (EEGH) of Gossypium herbaceum were dissolved in 1% alkaloids, flavonoids, glycosides, saponins, tannin and w/v Tween 80, to give 10, 20, 40, 60, 80and 100 mg/ml carbohydrate. (Table 1) respectively. Albendazole was used as the standard at 1, 2, 4, 6, 8 and 10mg/ml.10 ml of freshly prepared each Anthelmintic activity
extracts of Gossypium herbaceum, standard solution and The results of anthelmintic activity are shown in Table 2 1% v/v tween 80 were poured into petridish. The worms and Figure 1-3. The ethyl ether and Ethanolic extracts were washed with saline and released into the petridish showed significant paralysis as well as death time at all and the time taken for the worms to get paralyzed and concentrations. N-Hexane has no anthelmintic activity killed was noted. Observations were made for the time from 10 mg/ml to 60 mg/ml but 80 & 100 mg/ml shows taken for paralysis was noted when no movement of any effect but it is not significant when compared to standard sort could be observed except when the worms were drug albendazole. Among all the concentrations evaluated shaken vigorously. Time for death of worms were for the EEEGH 10, 20, 40, 60, 80 & 100 mg/ml solution, recorded after ascertaining that worms neither moved the 60, 80 and 100 mg/ml exhibited the most potent when shaken vigorously nor when dipped in warm water anthelmintic activity when compared with the positive control albendazole 10mg/ml. The 40 mg/ml of EEEGH revealed as like as standard. The same manner EEGH Statistical analysis
shows potent anthelmintic activity at the concentration of Experimental data are expressed as mean±standard error 80 and 100 mg/ml. The 60 mg/ml showed equal to of mean (SEM). Statistical analysis was performed by standard but the ethyl ether extract is showed more potent one-way ANOVA followed by Dunnett's method of anthelmintic activity than ethanol extract of Gossypium multiple comparisons was employed using Graph pad herbaceum with less concentration compared to ethanol Instat 3.0 software. Data were considered significant at Table 1: Phytochemical screening of methanolic and different fractions of Gossypium herbaceum
Steroids Alkaloids Glycosides Saponins Flavonoid Tannin Carbohydrates
C. Velmurugan,et al / Int. J. of Pharmacology and Clin. Res. Vol-1(1) 2015 [1-7]
Table 2: In vitro anthelmintic activity leaves of various extracts of Gossypium herbaceum linn.
Time taken for paralysis in
Time taken for death in
Results are expressed as mean ± S.E.M. (n=6) for each group; significance at p< 0.05, as compared to Standard Figure 1 Paralysis and death time of N-Hexane extract of Gossypium herbaceum compare to Albendazole.
death time in min.
Time of paralysis in min.
Alb 10 NHEGH NHEGH mg/ml mg/ml mg/ml mg/ml mg/ml Figure 2: Paralysis and death time of ethyl ether extract of Gossypium herbaceum compare to Albendazole.
C. Velmurugan,et al / Int. J. of Pharmacology and Clin. Res. Vol-1(1) 2015 [1-7]
Time of death in min.
Time of paralysis in min.
Figure 3: Paralysis and death time of ethanol extract of Gossypium herbaceum compare to Albendazole.
Death time in min.
Time of paralysis in min.
ethanol extracts showed presence of glycosides, alkaloids, The predominant effect of albendazole on Primary action flavanoids, tannins and phenolic compounds. Earlier is binding to beta tubulin and thus inhibition of studies proved that the polyphenolic compounds show microtubule polymerisation. The action is more specific anthelmintic activity (Niezen et al 1995) and some to parasitic beta-tubulin than that of host. Immobilisation synthetic phenolic anthelmintics e.g. niclosamide, & death of parasites occur slowly and they produce many oxyclozanide and bithionol are shown to interfere with biochemical changes in susceptible Nematodes Inhibition energy generation in helminth parasites by uncoupling of mitochondrial fumaratereductase, reduced glucose oxidative phosphorylation (Martin 1997). polyphenols transport and uncoupling of oxidative phosphorylation. from bryophytes were shown to have anthelmintic activity The ethyl ether and ethanolic extract of Gossypium against Nippostrongylus brasiliensis (Gamenara et al herbaceum demonstrated paralysis as well as death of 2001). Another possible anthelmintic effect of tannins is worms in a less time as compared to albendazole that they can bind to free proteins in the gastrointestinal especially at higher concentration of 100 mg/ml. While tract of host animal (Athnasiadou et al 2001) or hexane extract has shown no significant activity. glycoprotein on the cuticle of the parasite (Thompson et Preliminary phytochemical screening of hexane extract al 1995) and cause death. revealed the presence of steroids while ethyl ether and C. Velmurugan,et al / Int. J. of Pharmacology and Clin. Res. Vol-1(1) 2015 [1-7]
So the above point supported the anthelmintic activity of effective against parasitic infections of humans. Further, ethyl ether and ethanol extract of Gossypium herbaceum in future it is necessary to isolate the possible active might be due to phytoconstituents like tannins and phytoconstituents responsible for the anthelmintic activity phenolic compound once again it was confirmed by the mainly phenolic compound and study its pharmacological hexane extract was not shown significant activity it may be due to absence of phytoconstituent like phenolic compound. The mechanism of EEEGH and EEGH interfere with polymerization of tubulin and energy The authors are thankful to Chairman and Principal, Sri production in helminthiasis. Krishna Chaithnya College of Pharmacy, Madanapalle, Andhra Pradesh (India) for providing facilities to carry CONCLUSION
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