Antioxidant and Antibacterial Investigations on Essential Oils And

Natural Product Radiance, Vol. 6(2), 2007, pp.114-121 Research Article Antioxidant and antibacterial investigations on essential oils and acetone extracts of some spicesÜ Gurdip Singh1*, Sumitra Maurya3, Palanisamy Marimuthu4, H S Murali2 and A S Bawa2 1Chemistry Department, DDU Gorakhpur University, Gorakhpur- 273 009, Uttar Pradesh, India 2Defence Food Research Laboratory, Mysore- 570 011, Karnataka, India 3 Agharkar Research Institute, Pune-411 004, Maharashtra, India 4 Indian Agricultural Research Institute, New Delhi- 110 012, India †Part 49 *Correspondent author, E-mail: [email protected] Phone: 91-551-2200745 (R) 2202856 (O); Fax: 91-551-2340459; Received 7 April 2006; Accepted 11 September 2006 Abstract The studies on antioxidant and antibacterial potential of essential/volatile oils and acetone extracts of various spices are presented in this paper. The antibacterial activity of the volatile oils and acetone extracts of anise, ajwain, tejpat, Chinese Cassia bark, fennel, coriander, dill, turmeric and star anise have been studied against Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Bacillus cereus, B. subtilis and Staphylococcus aureus by disc diffusion and plate count methods. The results showed that volatile oils and extracts varied in their bioactivity. The volatile oils of ajwain, tejpat, Chinese Cassia bark and coriander were found to possess Ajwain excellent activity against all the Gram positive and Gram negative bacterial strains tested. These volatile oils and extracts are equally or more effective when compared with standard antibiotics diffusion and dilution of the test substance even at very low concentration. However, the acetone extract was found to be less effective as compared to volatile oils. Antioxidant activity of the oils and extracts were studied by DPPH, in a microbiological medium. Both reducing power, conjugated diene and chelating effect assays. They exerted concentration dependent microbiostatic5 and microbiocidal6 effects antioxidant activity in all the tested assays. are reported in the literature, although, Keywords: Spices, Volatile oils, Antimicrobial activity, Acetone extract, Antioxidant activity. there are discrepancies about the spectrum of activity and potency of individual plant IPC code; Int. cl.8 — C11B 9/00, A23L 1/22, A61K 36/00, A61P 17/18, A61P 31/04 extracts and essential oils. The antioxidant and antibacterial activity of essential oil isolated from higher plants especially from Introduction and acetone extract of some spices like spices in the form of essential oils have Natural products and naturally black cumin, black pepper, mace, etc. been extensively investigated3. Essential derived components from plants have have been earlier reported by us7. In this 1, 2 oils and extracts are heterogeneous group applications in controlling pathogens paper also, we report the antimicrobial of complex mixtures of organic in foods. The antioxidant and antibacterial and antioxidant potential of essential oils substances whose quality and quantity vary property of added material is very and extracts isolated from some more with growth stages, important to improve the shelf life of food spices, viz. anise, ajwain, tejpat, Chinese ecological conditions and material and at the same time provide Cassia bark, fennel, star anise, turmeric, other factors of the plant safety to consumers. The challenge is to dill and coriander. isolate, purify, stabilize and incorporate from which it is extracted4. The volatility and poor natural antioxidants and antimicrobials Materials and Methods into foods without adversely affecting solubility of essential oils Plant material sensory, nutritional and safety and extracts are The spices listed in Table 1 were characteristics. With the passage of time, problematic, particularly procured from local wholesale market of various active fractions or constituents with methods that rely on Tejpat 114 Natural Product Radiance Research Article Gorakhpur, during June to November, and three Gram negative, 2004 and voucher specimens were Escherichia coli, Salmonella deposited at the Herbarium of Science typhi and Pseudomonas faculty of DDU Gorakhpur University, aeruginosa were selected for the Gorakhpur. present study. The pure cultures of these bacteria were obtained from Coriander Star anise Isolation of volatile oil Microbiology Department of Defence The spices were powdered (800 Antioxidant assays Food Research Laboratory, Mysore, mesh size) using domestic model grinder DPPH and Reducing power assays India. They were subcultured on nutrient and were subjected to hydrodistillation in The DPPH (2, 2'-diphenyl-1- agar broth. a Clevenger type apparatus for 3 hours. picrylhydrazyl radical) assay was carried The volatile oils thus obtained were dried out as described by Cuendet and his co- Antibacterial screening over anhydrous sodium sulphate to remove workers8. The reducing power was Two different methods disc traces of moisture and stored in determined according to methods diffusion and plate count were employed refrigerator in dark at 4°C until use. reported earlier by Oyaizu9. for the determination of antimicrobial activities. The screening of spices showing Isolation of acetone extract Chelating effect of ferrous ions activity was done using method reported After the isolation of volatile oil, and Conjugated diene assays earlier by us7. the powdered spice materials were Chelating effect was determined dried at 45°C. The extract was according to the method of Shimada Statistical analysis obtained by extracting 20g of dried et al10. The antioxidant activity was also For the essential oil or acetone powder of spice with 900 ml acetone for determined by the conjugated diene extarct, three samples were prepared for 3 hours in a Soxhlet apparatus. The extract method as reported by Lingnert et al11. assays of every antioxidant and was concentrated up to 10 ml. The antibacterial attribute. The data were remaining acetone was evaporated by Antibacterial investigations presented as mean ± standard deviation placing the samples in a vacuum drier Tested microorganisms of three determinations (data were not under reduced pressure. The viscous Six pathogenic bacteria, three shown). Statistical analyses were extracts were stored in a refrigerator at Gram positive, Bacillus cereus, B. performed using a one-way analysis of 4oC until use. subtilis and Staphylococcus aureus, variance12. A probability value of P <0.05 was considered significant. Table 1 : Spices, parts and their major components used in the present study Common name Botanical name Family Parts used Major components Anise Pimpinella anisum Linn. Apiaceae Seed Anethole Ajwain Trachyspermum ammi (Linn.) Sprague Apiaceae Seed Thymol Tejpat Cinnamomum tamala Nees & Eberm. Lauraceae Leaf Eugenol Chinese Cassia bark Cinnamomum aromaticum Nees syn. Lauraceae Bark Cinnamaldehyde C. cassia Blume Fennel Foeniculum vulgare Mill. Apiaceae Seed Anethole Star anise Illicium verum Hook. f. Magnoliaceae Fruit Anethole Turmeric Curcuma longa Linn. Zingiberaceae Rhizome Ar-turmerone Dill Anethum graveolens Linn. Apiaceae Seed Carvone Coriander Coriandrum sativum Linn. Apiaceae Seed Linalool Vol 6(2) March-April 2007 115 Research Article Results and Discussion the volatile oils of star anise, dill, fennel has been reported to possess coriander and turmeric were also found bacteriostatic effect by Smith-Palmer 22 Antimicrobial studies to be highly active against all tested et al . Essential oils from dill The data reported in Tables 2 and microorganisms except Salmonella (Anethum graveolens Linn.), 3 showed excellent activities against the typhi, whereas their extracts exhibited coriander (seeds of Coriandrum test bacteria. Using the disc diffusion some activity though lesser than volatile sativum Linn.), cilantro (leaves of method (Table 2), the volatile oil of oils. The data was found to be highly immature C. sativum) and eucalyptus ajwain showed a broad spectrum of significant (P>0.05). (Eucalyptus dives) were also tested inhibition against all tested organisms. It The oils and extracts though less against Gram positive and Gram negative 3 23 is interesting to note that it causes effective at lower concentration, gave rise bacteria .Recently, Gupta et al reported complete inhibition of B. subtilis only to complete inhibition of bacterial growth antimicrobial activity of garlic, ginger, at 10µl dose of the oil and more than when their concentration increased. The carrot and turmeric pastes against E. coli. 80% mycelial zone inhibition was volatile oils and extracts are equally or obtained at 6µl. Moreover, a complete more effective when compared with Antioxidant studies zone of inhibition was obtained using dill standard antibiotics, at very low DPPH and Reducing power volatile oil against B. subtilis at 10 µl concentrations. Individual compounds The scavenging effect of the dose and more than 40% at 6 µl. The derived from volatile oils such as anethole essential oils and acetone extracts on DPPH volatile oil of cinnamon was found to be from anise, fennel and star anise, thymol radical linearly increased with increasing highly effective against E. coli, P. from ajwain, eugenol from tejpat, concentration (Table 4). All the acetone aeruginosa, B. subtilis and cinnamaldehyde from Chinese Cassia extracts exhibited strong radical scavenging Staphylococcus aureus as more than bark, ar-turmerone from turmeric, carvone activity in comparison with commercial 40% zone inhibition was obtained at 10 from dill and linalool from

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