Saroj Kumar Raul et al. / Journal of Pharmacy Research 2012,5(7),3813-3814 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info An in-vitro evaluation of the anthelmintic activity of extracts of officinalis, Zingiber zerumbet and Curcuma longa, a comparative study *Saroj Kumar Raul, Gopal K Padhy, Jasmin P Charly, K Venkatesh Kumar Maharajah’s College of Pharmacy,Phoolbaugh,Vizianagaram,Andhra Pradesh, India Received on:07-04-2012; Revised on: 12-05-2012; Accepted on:16-06-2012

ABSTRACT The present study was undertaken to evaluate the Anthelmintic activity of ethanol extracts of Zingiber officinalis (Zingiberacae), Zingiber zerumbet (Zingiberacae) and Curcuma longa on Indian earthworm, Pheretima posthuma.Various concentrations (10ug/ml, 25ug/ml, 50ug/ml) of all extracts were tested and results were expressed in terms of time of paralysis and time of death of the earthworm. Piperazine citrate (10ug/ml) was used as reference standard and sterile water as control group.

KEYWORDS: Anthelmintic activity, Pheretima posthuma, Piperazine citrate, Zingiber officinalis, Zingiber zerumbet, Curcuma longa

INTRODUCTION: Medicinal used in India for centuries have an important therapeutic Curcuma longa commonly known as is a rhizomatous herbaceous source for treating a variety of ailments and has been found to immense global perennial of the family, . [2]. It is native to tropical importance. India is perhaps the largest producer of medicinal herbs and is South Asia and needs temperatures between 20 °C and 30 °C and a consider- rightly called the ‘Botanical Garden of the world’. Medicinal herbs have been able amount of annual rainfall to thrive.[3] Plants are gathered annually for used for thousands of years in the indigenous system of medicines like their , and propagated from some of those rhizomes in the following , Siddha and Unani. season. Turmeric is currently being investigated for possible benefits in Alzheimer’s disease, [10] cancer, [11][12] arthritis, and other clinical disor- Helmenthiasis is one of the most common infections in mankind affecting a ders.[13][14] As an example of preliminary laboratory research, turmeric large proportion of the world population. Parasitic diseases may cause se- ameliorated the severity of pancreatitis-associated lung injury in mice.[15] vere morbidities including lymphatic filariasis and onchocerciasis. Develop- ment of resistance to most of the commercially available anthelmintics be- Curcuma longa and Zingiber officinale rhizomes are rich for phytoconstituents comes a problem world-wide. viz. alkaloids, saponins, flavonoids, terpenes and steroids. These drugs are widely used in the treatment of different ailments in the Indian system of Zingiber officinale commonly known as ginger lends its name to its genus and medicine. family Zingiberaceae. Ginger produces clusters of white and pink flower buds that bloom into yellow flowers. Because of its aesthetic appeal and the MATERIALS AND METHODS: adaptation of the plant to warm climates, ginger is often used as landscaping The methodology adapted to evaluate the anthelmintic activity of the rhi- around subtropical homes. It is a perennial reed-like plant with annual leafy zomes of Zingiber offficinale, Zingiber zerumbet and Curcuma longs is as stems, about a meter (3 to 4 feet) tall. Traditionally, the root is gathered when follows the stalk withers; it is immediately scalded, or washed and scraped, to kill it and prevent sprouting. Plant materials: The rhizomes of Zingiber officinale, Zingiber zerumbet and Curcuma longa Preliminary research indicates that nine compounds found in ginger may bind were collected from the local market of Vizianagaram, Andhra Pradesh. They to human serotonin receptors, which may explain ginger’s extensive effects were identified and confirmed from the Department of Botany in Maharajah’s on the GI tract and suggesting a mechanism for its effects on anxiety. [7] Autonomous College, Vizianagaram, Andhra Pradesh.

Zingiber zerumbet also known as shampoo ginger belongs to the family Collection of worms: Zingiberaceae. It is a vigorous ginger with leafy stems growing to about 1.2 m The Indian earthworm Pheretima posthuma were collected from the water tall. It is found in many tropical countries. The rhizomes of Zingiber zerumbet, logged area of Dharmapuri, Vizianagaram, Andhra Pradesh. They were iden- have been regularly used as food flavoring and appetizer in various cuisines tified from the Department of Zoology, Maharajah’s Autonomous College, while the rhizomes extracts have been used in Malay traditional medicine to Vizianagaram, Andhra Pradesh. treat various types of ailments like inflammatory and pain-mediated dis- eases, worm infestation and diarrhoea.[1] Preparation of extract: The collected materials were washed thoroughly in water, cut into small pieces and air-dried for 3 weeks at 35-40 degree centi- grade. Extraction was done by using Soxhlet apparatus with 90% ethanol as *Corresponding author. solvent. The extracts were concentrated under reduced pressure, dried, pow- dered and stored in an air-tight container. Saroj Kumar Raul, Maharajah’s College of Pharmacy, Preparation of test sample: Phoolbaugh,Vizianagaram, Samples for in-vitro study were prepared by adding Tween 80 as suspend- Andhra Pradesh, India ing agent in 2.5 gram of each extract in 25 milliliters of distilled water to obtain

Journal of Pharmacy Research Vol.5 Issue 7.July 2012 3813-3814 Saroj Kumar Raul et al. / Journal of Pharmacy Research 2012,5(7),3813-3814 a stock solution of 100mg/ml. From that stock solution different working ralysis and 16.50 min. for death. While in Zingiber officinale for extract at the dilutions were prepared to get concentrations of 10ug/ml, 25ug/ml and 50ug/ concentration of 10mg/ml, the time of paralysis and death was found to be ml. 13.20 min. and 32.40 min. respectively. At concentration of 20 mg/ml, it was 7.50 min. for paralysis and 24.20 min. for death. For concentration at 50mg/ Anthelmintic activity: ml, the time of paralysis and death was 6.40 min. and 17.5 min. respectively.In The Anthelmintic activity was performed according to the methodology case of combination of both the plant extracts (Curcuma longa and Zingiber followed by Ajayieoba et.al, 2001[6] on adult Indian earthworm, Pheretima officinale), the time of paralysis and death was 10.6 min. and 31.8 min. posthuma due too it’s anatomical and physiological resemblance with human respectively at concentration of 10 mg/ml. At concentration of 20 mg/ml, the intestinal roundworm parasite. Earthworms were divided into 3 groups; time of paralysis and death was 6.8 min. and 27 min. respectively and at 50 consisting of 6 earthworms in each group and were released into 20ml of the mg/ml concentration, the time of paralysis and death was 4.8 min. and 16.3 desired formulation. Group I serve as control and receive only distilled wa- min. respectively. The observations with piperazine citrate showed that ter, Group II serves as standard and receives standard drug Piperazine citrate time of paralysis and death was 7.2 min. and 15.9 min. respectively for (10mg/ml), Group III serves as the alcoholic extract of different concentra- concentration at 20 mg/ml. It was observed that both ginger as well as tur- tions. meric hydro-alcoholic extracts showed a remarkable anthelmintic potential against intestinal parasitism. Amongst the both extracts, Curcuma longa showed Observations were done for the time taken for the paralysis and death of better activity in combination with Zingiber officinale. Anthelmintic activity individual worms. Paralysis was said to be occurred when there no move- of Curcuma longa was enhanced in the presence of Zingiber officinale. ment of any sort could be observed except that the worms were shaking vigorously. Death was said to be occurred when the worms lost their motility REFERENCES: followed with fading of their body color. 1. Anthelmintic activity of medicinal plants with particular reference to their use in animals in the Indo-Pakistan subcontinent by M S RESULTS AND DISCUSSION: Akhtar, ZafarIqbal, M N Khan, Muhammad Lateef, Small As shown in the Table 1 below the data revealed that Curcuma longa extract Ruminant Research (2000),Volume: 38, Issue: 2, Pages: 99-107 at the concentration of 10 mg/ml showed the time of paralysis and death at 2. Medicinal plants for helminth parasite control: facts and fiction ,S. 12.40 min. and 34 min. respectively. For concentration of 20mg/ml, the Athanasiado, J. Githiori and I. Kyriazakis, Animal (2007), 1:9, pp paralysis and the death time was found to be 7.20 min. and 23.40 min. 1392–1400 respectively. At the concentration of 50mg/ml, time was 5.40 min. for pa- 3. Jain S, Shrivastava S, NayakS and Sumbhate S, Plant Review. Recents trends in Curcuma longa Linn. Phcog. Rev. 2007; 1:119- Table:1 Anthelmintic activity of Extracts 128 4. Ajayieoba EO, Onocha PA, Olarenwaju OT, In-vitro anthelmintic Test Subject Concentration Time taken for Time taken for properties of Buchholziacoriaceae and Gynandropsisgynandra (mg/ml) paralysis(P) in mins death(D) in mins extract. Pharm. Biol. 2001; 39:217-20. Vehicle - - - 5. Identification of serotonin 5-HT1A receptor partial agonists in

Piperazine citrate 10 22:40 ±1.5 46.50 ± 2.4 ginger. Nievergelt A. Huonker P. Schoop R. Altmann KH. Gertsch J. Bioorganic & Medicinal Chemistry. 18(9):3345-51, 2010 May 1 6. Iqbal Z, Nadeem QK, Khan MN, Akhtar MS, Waraich FN. In vitro Curcuma longa 10 12.40 ± 1 34 ± 2 25 7.20 ± 1 23.40 ±1.5 anthelmintic activity of Allium sativum, Zingiber officinale, 50 5.40 ± 1 16.50 ± 1 Curcurbita mexicana and Ficusr eligiosa .Int J AgrBiol 2001; 3:454-

Zingiber officinale 10 13.20 ± 1 32.40 ± 2 7. 25 7.50 ±1 24.20 ± 1,5 7. T.Ghosh, T.k.Maity, A.Bose and G.K. Dash, Anthelmintic activ- 50 6.40 ± 1 17.5 ± 1.5 ity of Bacopa monierri, Indian J.Nat Prodct, 2005, 21, p-16-19

Zingiber zerumbet 10 108 ± 0.9 165.40 ± 1.5 8. Chatterjee K.D, Parasitology and helminthology, Cuha Ray 25 85 ±1.2 135.40 ± 0.9 SreeSaraswathyPress Ltd. Calcutta. 1967, pp. 168-169 50 60 ±1.1 110.20 ± 1.3 9. Vigar Z, Atlas of Medicinal of Parasitology, P.G.Publishing house,

Zingiber officinale 10 11.2 ±1 31.40 ± 1.6 Singapore; 1984, P.No: 216. + 25 7.2 ± 1 22 ± 1 10. Waller PJ, The future of anthelmintics in sustainable parasite con- Cucuma longa (1:1) 50 5.60 ± 1 15 ± 1 trol programme for livestock. Helminthologia. 2003

Zingiber zerumbet 10 42 ± 1.5 75.20 ± 0.7 11. Mascolo N, Jain R, Jain SC, and Capasso F, Ethnopharmacologic + 25 36 ± 1.2 46.10 ± 0.5 Investigation of ginger (Zingber officinale). Journal of Curcuma longa (1:1) 50 30 ± 0.8 40 ±1.1 Ethnopharmacol. 1989; 27:129-140. All values represent Mean +/- SD,n=6 in each group Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.5 Issue 7.July 2012 3813-3814