ANTI-TERMITE ACTIVITY OF HEARTWOOD OF DALBERGIA SISSO ROXB. EX. DC.

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WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES Harsha et al. SJIF Impact Factor 2.786 Volume 3, Issue 6, 673-679.

*Harsha Kharkwal 1, Amit Kharkwal 2, Preeti Panthari 3, Harendra Kharkwal 4 1* Amity Center for Carbohydrate Research, J-1 Block, LG-14, Amity University Uttar Pradesh, Noida, 201301.

3 Amity Institute of Phytochemistry and Phytomedicine, J-1 Block, LG-14, Amity University Uttar Pradesh, Noida. 4 Ministry of Environment and Forest, Govt. of India, CGO Complex, New Delhi- 110003.

1 WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES Harsha et al. SJIF Impact Factor Volume 3, Issue 6, Research Article ISSN ANTI-TERMITE ACTIVITY OF HEARTWOOD OF DALBERGIA SISSO ROXB. EX. DC. *Harsha Kharkwal 1, Amit Kharkwal 2, Preeti Panthari 3, Harendra Kharkwal 4 1* Amity Center for Carbohydrate Research, J-1 Block, LG-14, Amity University Uttar Pradesh, Noida, Amity Institute of Microbial Technology, E-3 Block, 4 th Floor, Amity University Uttar Pradesh, Noida. 3 Amity Institute of Phytochemistry and Phytomedicine, J-1 Block, LG-14, Amity University Uttar Pradesh, Noida. 4 Ministry of Environment and Forest, Govt. of India, CGO Complex, New Delhi Article Received on 25 March 2014, Revised on 17 April 2014, Accepted on 10May 2014 *Correspondence for Author Dr. Harsha Kharkwal Amity Center for Carbohydrate Research, J-1 Block, LG-14, Amity University Uttar Pradesh, Noida, ABSTRACT Objective: The present study aimed to study the termite repellency of the crude extract of heartwood of Dalbergia sisso Roxb. ex DC. tree using several solvents. Method: The extraction was done according to the standard method of Soxhlet extraction followed by Column chromatography and Anti termite activity. Result: The chloroform extract possesses highest potential which was further fractionated. 0.2% concentration of a fraction Chloroform: Methanol (99:1) showed anti-termite activity up to 92%. Conclusion: The plant extracts can be used as an alternative for synthetic pesticides for termite control in buildings. Key words: Ethnomedicinal uses, Termite repellency, anti-termite activity. INTRODUCTION Dalbergia sisso belongs to family Fabaceae which is commonly known as Sissam in Hindi is known worldwide as timber species. It is found throughout the country but large quantity of sisso timber is available from North India. The tree has immense enthnomedicinal uses and has been used traditionally since a long time. It is known as premier timber species of rosewood genus. It is a folk remedy for excoriations, gonorrhea and skin ailments [1]. In Ayurveda the wood and bark have been described as abortifacient, anthelmintic, antipyretic, Vol 3, Issue 6,

2 aperitif, aphrodisiac, expectorant and refrigerant, while the leaf juice is advised for eye ailments [2]. The wood and bark used for anal disorders, blood diseases, burning sensations, dysentery, dyspepsia, leucoderma and skin ailments. Yunani use the wood for blood disorders, eye & nose disorders scaling urine, stomach problems and syphilis. Leaves decoction is used in the gonnorhoea treatment whereas leaf mucilage with sweet oil is used in excoriations [1]. The leaves possess antipyretic and analgesic activites [3] while do not possess anti-bacterial, anti-protozoal, and antiviral activity but is antidiarrhoeal [4]. Lots of formulation using bark, leaves and nector are used in treating a number of ailments [1]. The ethanolic extract of the bark possess anti-inflammatory activity [5]. The anti-inflammatory activity of root was carried out in rats and it was found that the inflammation was reduced in rat with hind paw [6]. In a comparative study done on ethanolic extract of Dalbergia sissoo bark and glibenclamide a hypoglycemic drug, it was found that the plant extract possess potent antidiabetic activity [7]. Traditionally the plant has been used as timber or fire wood and most specially treating various ailments [8]. The application in the treatment of lots of digestive disorders and skin diseases are well known [1, 3]. Cyclodiene hydrocarbons in the human body are hugely responsible for the danger of long term toxicity. Such toxicity resulted in national and international legislation to limit and even completely ban the use of certain compounds of this group. These chemicals are extensively used to control pest in agriculture, animal and household applications. There is an urgent need to control the use of bio-agents which are less toxic and more readily degradable. They possess certain advantages over synthetic insecticides because of their non-toxic and ecofriendly characteristics and are eco-friendly compounds. A large number of plants have been reported to possess pest controlling activity and there is much scope to search more alternative from plant wealth [9]. Plants are the richest source of bioactive organic chemicals on earth. They are the store house of secondary metabolites such as alkaloids, terpenoids, steroids, flavonoids, etc. which may act as toxicants, repellant, plant toxicant and frequently lethal to insect upon the contact or their vapour may kill the insect by fumigation while repellent or deterrent causing insect to avoid contact with target species. Termite is a pest which causes considerable damage to wood, clothes, paper etc. They belong to insect order Isoptera [10]. They are soft bodied and medium sized and may be classified into two main groups (1) Wood dwellers and (2) ground dwellers. The natural food of termite is cellulose which is a major compound of cell walls of Vol 3, Issue 6,

3 trees or plants. Some of the toxicant like creosote, coal tar and carbolic acid are used but these have limited effects. Synthetic toxicant are much affective but they cause long term toxicity to living beings and damage to environment. Promotion of bio-toxicant is an appropriate solution and a need of hour. The use of medicinal plant extracts having termiteresistance property are good alternative of synthetic pesticides [10]. MATERIAL AND METHODS The heartwood of the tree D. sisso was collected and dried in shade and crushed through Electric Shredder. The crushed material (2 kg) has been extracted with ethyl alcohol in soxhlet apparatus for five days. The extract was concentrated in Rota-vapour at reduced pressure and the aqueous emulsion was extracted with n-hexane (4x100 ml) followed by successive washing. The solvent was evaporated under reduced pressure and dried (yield gm). The remaining water soluble portion was again extracted with chloroform (5x100 ml) and n-butanol (5x100 ml), washed with water they dried. Solvent was evaporated under reduced pressure so that gm of gummy chloroform extract, gm of solid n-butanol extract and gm of water soluble extract were obtained. Column Chromatography of Chloroform Extract The gm of gummy residue was subjected to column chromatography over 800 gm silica gel packed in n-hexane. 10 fractions of 200 ml were collected in the following order as shown in Table 1. All the fractions were concentrated under vacuum and dried. Each fraction was subjected to the testing of anti-termite activity. Determination of Termite Repellency 18 petridishes of 90 mm diameter were cleaned, washed and dried in oven for 2 hours. Thereafter 90 mm diameter Whatman filter papers (6 replicas for each extract) were weighed without loading the material. Whatman filter papers were folded into half for repellency test of termites. Now the half portion of the filter papers were dipped into the plant extract. The rest half remains untreated. These filter papers were dried at room temperature. The half loaded filter papers were again weighed and the loaded material was calculated. For the present studies, the active workers of termite species Microcerotermes beesonii were selected. Termite species 25 in number were released on each filter paper kept in the petridishes. The total numbers of live termites moving towards untreated area were observed after every 2 hours till 24 hours. Total numbers of dead termites were also observed. Percentage repellency was calculated by following formula: Vol 3, Issue 6,

4 UT % repellency = x 100 UT + T Where UT= Untreated part of filter paper T= Treated part of filter paper RESULTS The work reported in this paper has been carried out on plant products with a view to find out alternative to highly toxic and non-degradable synthetic pesticides. Heartwood of D. sisso was crushed and extracted in different solvents. Six replicas of each extract were used. Mean repellency was calculated in all the three extracts and thus the repellent power of the plant extract calculated. The observations on termite repellency of the extracts are recorded in Table 2-6. Crude extract in n-hexane, chloroform and n-butanol were studied for termite repellency as shown in Table 2-4. The extracts showed repellency varying from 12-18%. Further, various Fractions of crude extracts obtained in chloroform were separated by column chromatography as presented in Table-1. The fractions obtained in CHCl 3 :MeOH (99:1) and CHCl 3 :MeOH (95:5) were studied for termite repellency. As shown in Table-5, the fraction obtained in CHCl 3 :MeOH (99:1) showed repellency up to 92% in solution of 0.2% concentration. However, at higher concentration the repellency was low. Similarly as shown in Table-6, the fraction obtained in CHCl 3 :MeOH (95:5) showed highest termite i.e. 84% at 0.4% concentration. However, in this case also the repellency was low at higher concentrations. The reason of lowering of termite repellency at higher concentration needs study in depth. Table 1: Chromatographic Fractions with Different solvent systems Solvent System 1. n-hexane 2. n-hexane: chloroform (20:1) 3. n--hexane: chloroform (10:1) 4. n-hexane: chloroform (5:1) 5. n-hexane: chloroform (3:1) 6. n-hexane: chloroform (1:1) 7. n-hexane: chloroform (1:2) 8. Chloroform 9. Chloroform: methanol (99:1) 10. Chloroform: methanol (95:5) 11. Methanol Vol 3, Issue 6,

5 Table 2: Observations on repellent activity with crude-1 (n-hexane extract) Table 3: Observations on repellent activity with crude-2 (Chloroform extract) S. No. Solution (%) Total number of Termites Number of termites repelled Repellency (%) Table 4: Observations on repellent activity with crude -3 (n-butanol extract) Table 5: Observations on repellent activity of fraction obtained in Chloroform: methanol (99:1) Vol 3, Issue 6,

6 Table 6: Observations on repellent activity of fraction obtained in Chloroform: methanol (95: 5) DISCUSSION There is an enhance in the anti-termite activity seen when the n-hexane, chloroform and n- butanol extract are taken as such, the highest in the case of n-hexane leading to the probability of the anti-termite activity from the oil of Dalbergia sisso. However, when the elutes where taken of chloroform fraction along with methanol there was almost 3 times anti termite activity observed with the maximum being 92 %. The results show the efficacy of this commercially important tree to be further utilized by formulating it for anti termite action. CONCLUSION The study reveals that the plant extracts may have good prospects as an alternative for synthetic pesticides for termite control in buildings. ACKNOWLEDGEMENTS Authors are thankful to Dr. Ashok K Chauhan, Founder President, Ritanand Balved Educational Foundation for support. They are also thankful to Shri. Atul Chauhan, Chancellor, Amity University Uttar Pradesh and Prof. Dr. (Mrs.) Balvinder Shukla, Vice Chancellor, Amity University Uttar Pradesh for their constant support and guidance. We are thankful to the MoEF for Financial assistance. (Grant No. AUUP/2010/164) REFERENCES 1. Lal HS, Singh S. Ethnomedicinal uses of Dalbergia sissoo Roxb in Jharkhand. Int J Ayur Herb Med 2012; 2(1): Hajare SW, Chandra S, Sharma J, Tandon SK, Telang AG. Anti-inflammatory activity of Dalbergia sisso leaves. Fitoterapia 2001; 72: Vol 3, Issue 6,

7 3. Hajare SW, Chandra S, Tandan SK, Sharma J, Lal J, Telang AG. Analgesic and Antipyretic activites of Dalbergia sissoo leaves. Indian J Pharmacol 2000; 32: Brijesh S, Daswani PG, Tetali P, Anita NH, Birdi TJ. Studies on Dalbergia sissoo (Roxb.) leaves: Possible mechanism(s) of action in infectious diarrhoea. Indian J Pharmacol 2006; 38(2): Asif M, Kumar A. Anti-Inflammatory activity of ethanolic extract of Dalbergia sissoo (Roxb.) bark. Malays J Pharma Sci 2009; 7(1): Kumar SM, Kumud U. Anti-inflammatory activity of root of Dalbergia sissoo (Rox.b) in Carrageenan-induced paw edema in rats. Phcog J 2010; 2(11): ) 7. Pund KV, Vyawahare NS, Gadakh RT, Murkute VK. Antidiabetic evaluation of Dalbergia sissoo against alloxan induced diabetes mellitus in wistar albino rats. J. Nat.Prod Plant Resour 2012; 2(1): Ministry of Health & Family Welfare. The Ayurvedic Pharmacopoeia of India. Part I, Volume III. Department of Indian Systems of Medicine & Homeopathy; New Delhi: Govt. of India; Jaya V, Dubey NK. Prospectives of botanical and microbial products as pesticides of tomorrow. Banaras Hindu University. Varanasi India; Singh Y, Ranawat BS, Verma RK, Nayal SS. Termite control with medicinal plant products. J Med Aro Plant Sci 2001; (4A-1A): Vol 3, Issue 6,

In vitro study of antibacterial activity of Carissa carandas leaf extracts

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