Shivani Joshi et al. / Journal of Pharmacy Research 2011,4(7),2282 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Antibacterial and Antifungal Properties of Crude Extracts of asiatica L. Aerial Parts *Shivani Joshi1, Devendra Mishra2, K.S. Khetwal1 and Ganga Bisht1 1Department of Chemistry, D.S.B. Campus, Kumaun University, Nainital-263002, 2Department of Applied Chemistry, Birla Institute of Applied Sciences, Bhimtal, Nainital, India Received on: 12-04-2011; Revised on: 18-05-2011; Accepted on:21-06-2011

ABSTRACT Methanol, chloroform and aqueous extracts of Buddleja asiatica Lour. (Buddlejaceae) aerial parts obtained by soxhlet extraction method were screened for their antimicrobial activity against six bacteria (Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris,

Bacillus mycoides) and two fungi (Aspergillus niger, Candida albicans) using disc diffusion method. Out of the three extracts, CHCl3 extract exhibited significant activity against P. aeruginosa and B. mycoides while it showed best antifungal activity against A. niger. The other two extracts showed moderate antibacterial and antifungal activity for all the concentrations. The activity shown by the CHCl3 extract was found even higher than Chloramphenicol (25µg/ disc, diameter 5mm) against Pseudomonas aeruginosa and Fluconazole (25µg/ml) against Aspergillus niger.

Key words: Buddleja asiatica, Buddlejaceae, antibacterial, antifungal INTRODUCTION The genus Buddleja, comprises of approximately 100 species most of which extract was analyzed in triplicate. The antimicrobial activity of the extracts are and a few being trees1. Several types of chemical compounds like was determined by measuring the zone of inhibition (mm), including the diameter terpenoids, flavonoids, saponins, and lignans have been isolated from this of disc and the results (mm of zone of inhibition) were expressed as average genus2. Buddleja asiatica, commonly known as Butterfly Bush has been used values. as an abortifacient and also in the treatment of skin complaints.3,4 The juice of the is applied as a wash to treat skin diseases.5 A concentrated infusion of RESULTS AND DISCUSSION 6 the roots finds use as a treatment for malaria . A new compound, named buddlin Out of the three extracts, CHCl3 extract showed significant antibacterial and was isolated from the whole plant of Buddleja asiatica.2 Four non-phenolic antifungal activity upto 1000 µg/ml. It exhibited the best antibacterial activity antioxidant compounds were isolated from the leaves of Buddleja asiatica.7 against B. mycoides and P. aeruginosa while showed good antifungal activity Garg and Oswald (1981) have shown that the essential oil from the leaves has against A. niger. All the bacterial strains were found sensitive for water extract, however, against fungal strains, the activity was mild for A. niger and C. in vitro antifungal activity while no investigations on antimicrobial activity albicans was found resistant at all concentrations. Methanol extract exhibited of crude extract of B. asiatica are published so far. Hence, this study was very mild antibacterial and antifungal activity. The activity shown by the carried out to screen the crude organic extract (methanol and chloroform) and CHCl3 extract was found even higher than Chloramphenicol against aqueous extract of B. asiatica for antimicrobial activity. Pseudomonas aeruginosa and Fluconazole against Aspergillus niger.. The zone of inhibition markedly decreased on decreasing the concentration of the MATERIALS AND METHODS extracts for all the strains used for study (Table 1.). Plant Material The higher zone of inhibition shown by these extracts against tested organisms The aerial parts of the plant were collected in the month of February, 2009 revealed that they can be used as an alternative to standard antibiotics in the from Nainital, India and authenticated by Botanical Survey of India, Dehradun. treatment of infections caused by these microorganisms and since most of the A voucher specimen (No.112965) was deposited in the Herbarium Section at microorganisms are developing resistance against the known antibiotics, BSI, Dehradun, India. therefore, these can be the best therapeutics option as antimicrobial agents. Aerial parts of the plant were shade dried, powdered and extracted with methanol Table.1. Antimicrobial activity of Buddleja asiatica L. aerial parts using soxhlet apparatus. After extraction the filtrate was concentrated on a Zone of inhibition (mm) Standard rotary evaporator under vacuum at 20°C till a residual mass was obtained. The Extract (µg/ml) alcoholic residue was partitioned with chloroform/ water (50:50, v/v). Microorganism 4000 2000 1000 500 250 125 CP FZ Chloroform and water layers were separated with separating funnel and M C W M C W M C W M C W M C W M C W concentrated under reduced pressure. S. aureus 9 13 9 7 12 8 6 9 7 - 7 6 - 6 - - - - 12 - B. mycoides - 15 10 - 13 9 - 12 8 - 8 7 - 6 6 - - - 25 - Microorganisms B. subtilis - 8 10 - 7 9 - 6 8 - - 7 - 6 - - - 30 - Three gram positive, three gram negative and two fungi were used for P. vulgaris - 7 10 - 6 9 - 7 - - 6 - - - - - 14 - antimicrobial activity studies. Gram positive bacteria were Staphylococcus E. coli 7 8 11 6 7 8 5 6 7 - - 6 - - - - - 23 - aureus (MTCC 3160), Bacillus subtilis (MTCC 441) and Bacillus mycoides P. aeruginosa 8 16 10 7 12 9 6 11 8 - 9 7 - 7 6 - 6 - 10 - C. albicans 9 12 - 7 11 - - 9 - - 8 - - 7 - - 6 - - 15 (MTCC 645). Gram negative bacteria were Escherichia coli (MTCC 406), A. niger - 16 9 - 13 6 - 9 - - 8 - - 7 - - 6 - - 13 Pseudomonas aeruginosa (MTCC 424) and Proteus vulgaris (MTCC 426). Yeast like fungi used were Candida albicans (MTCC 227) and Aspergillus niger (MTCC 404). Required microorganisms were procured from Institute of M = Methanol Extract, C = Chloroform Extract, W = Water Extract, Microbial Technology, Chandigarh, India. CP = Chloramphenicol (25 µg/disc, 5mm in diameter), FZ= Fluconazole (25 µg/ml)

Determination of Zone of Inhibition ACKNOWLEDGEMENTS 9 The authors wish to thank Botanical Survey of India, Dehradun for plant The disc diffusion method was used to evaluate the antimicrobial activity. identification and Department of Pharmaceutical Sciences, Kumaun University, The test solutions of the MeOH, CHCl and water extract at the concentrations 3 Bhimtal for providing necessary facilities to carry out present investigation. of 4000, 2000, 1000, 500, 250 and 125 µg/ml were prepared by dissolving the extracts in dimethylsulphoxide (DMSO). 0.1ml of test solutions were injected into sterilized discs of 5mm in diameter. Chloramphenicol (25µg/disc) and REFERENCES 1. Houghton PJ. Ethnopharmacology of some Buddleja species. Journal of Ethnopharmacology, fluconazole (25µg/ml) were used as positive controls. As a negative control, a 1984, 11, p293-308. blank disc impregnated with DMSO was used. The test discs, standard discs and 2. Chen H, Xu C, Liu, DQ, An SQ, Tan RX. Buddlin, a New Compound from Buddleja asiatica. blank discs were placed in petridish with a particular microorganism. The Fitoterapia, 2005, 76, p588-89. petridishes were then incubated at 370C for 24h for bacterial growth and at 3. Chopra RN, Nayar SL, Chopra IC. Glossary of Indian Medicinal , CSIR, New Delhi, 1986 0 4. Anonymous. Medicinal Plants of , Department of Medicinal Plants, Nepal, 1993. 27 C for 48h for the growth of yeast. Nutrient agar and potato dextrose agar 5. Manandhar NP. “Plants and People of Nepal”, Timber Press, Oregon, 2002. medium were used for the growth of bacteria and yeast respectively. Each 6. Reis Altschul, S. von. Drugs and Foods from Little-known PIants, Harvard University Press, Cambridge, MA, 1973. 7. Sayed MM, Hameed SSA, Ahmed WS, Wakil EA. Non-Phenolic Antioxidant Compounds from *Corresponding author. Buddleja asiatica, Z. Naturforsch (C), 2008, 63, p483-91. Shivani Joshi 8. Garg SC, Oswald VB. In vitro antifungal activity of the essential oil of Buddleja asiatica, Riuista Ztuliana Essenze, Profumi, Piante Officinali, Aromi, Saponi, Cosmetici, 1981, 63, p365. Department of Chemistry, D.S.B. Campus, 9. Murray PR, Baron EF, Pfaller MA. Manual of Clinical Microbiology, Washington DC, ASM, 1995. Kumaun University, Nainital-263002,India Source of support: Nil, Conflict of interest: None Declared Journal of Pharmacy Research Vol.4.Issue 7. July 2011 2282