ijprbs 1013-1

See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/275343485

IJPRBS 1013-1 Dataset · April 2015

READS

72

1 author: Dr. Ebrahim Talebi Islamic Azad University 52 PUBLICATIONS 39 CITATIONS SEE PROFILE

All in-text references underlined in blue are linked to publications on ResearchGate, letting you access and read them immediately.

Available from: Dr. Ebrahim Talebi Retrieved on: 23 July 2016

Research Article CODEN: IJPRNK E. Talebi, IJPRBS, 2015; Volume 4(2): 215-224

ISSN: 2277-8713 IJPRBS

INTERNATIONAL JOURNAL OF PHARMACEUTICAL RESEARCH AND BIO-SCIENCE CHEMICAL CONSTITUENT, ANTIMICROBIAL AND ANTIOXIDANT ACTIVITIES OF ANVILLEA GARCINI-AN ESSENTIAL OIL PLANT M. H. FARJAM1, E. TALEBI2, SHIVAKUMAR3, I. NASROLLAHI2 1. Department of Chemistry, Firoozabad Branch, Islamic Azad University, Firoozabad, IRAN 2. Young Researchers and Elite Club, Darab Branch, Islamic Azad University, Darab, Fars, IRAN 3. Department of Studies in Sericulture Science, Manasagangotri, University of Mysore, Mysore – 570 006, Karnataka, INDIA

Accepted Date: 05/04/2015; Published Date: 27/04/2015 Abstract: The primary reason behind this research is to know the chemical constituents, antimicrobial and antioxidant activities of a popular essential oil plant namely, Anvillea garcini (A. garcini). In this investigation, the essential oils were extracted from the leaves, flowers and all the other parts of an A. garcini plant, through the standard hydro distillation method. The content of chemical composition of essential oil was observed and the data obtained this observation with (GC/MS) were clear showed that, 25 combinations derived from 97.44 percent of total essential oil. Further, the respected main constituents of this oil were myristicin (58/79%), Bicyclo (5.3.0) decane, 2-methylene-5-(1-methylvinyl)-8-methyl (7/71%), 5-Methyl-4-nonene (3/46% (E)-Ocimene (3/39%) recorded. Further, the essential Oil was showed inhibitory activity against gram-negative bacteria. Apart, antifungal activity against fungi like, Klebseilla oxytoca and Fusariumsolani (MIC=8) was clearly noticed. Furthermore Dependent on the free radical scavenging antioxidant activity of essential oil was DPPH (IC50=0.0209± 0.0028) showed against the antioxidant activity of essential oil was discussed here in this research work. Keywords: Anvillea garcini, hydro distillation, GC/MS, antimicrobial and antioxidant, DPPH

Corresponding Author: MR. E. TALEBI. Access Online On: www.ijprbs.com How to Cite This Article: PAPER-QR CODE

E. Talebi, IJPRBS, 2015; Volume 4(2): 215-224

Available Online at www.ijprbs.com

215



INTRODUCTION The usage of herbal medicines for different illness/diseases and for different age old population in Asia represents a long history of human interactions with the environment. It is accepted fact that, according to an investigation conducted by the World Health Organization (WHO), wherein more than 80% of the world's population relies on traditional medicine for their primary healthcare needs. Prior to that, a vast knowledge of how to use the plants/plant’s material against different illnesses may be expected to have accumulated in areas where the use of plants is still of great importance [1].Since, ancient times the crude herbal oil of aromatic plants have been in use for different purposes, such as food, drugs and perfumery [2]. Further, essential oils are a diverse group of natural products that are important sources of aromatic and flavoring chemicals in food, industrial, and pharmaceutical products respectively. Essential oils are largely composed of terpenes and aromatic poly propanoid compounds derived from the acetate-mevalonic acid and the shikimic acid pathways respectively [3] Moreover, A. garcini (Family Asteraceae) is a genus of small herbs or shrubs found in northern temperate regions [4]. A reinvestigation of a sample collected in the south of Iran (40 km north of the Persian Gulf). According to authors [5], Plants of the Asteraceae family are well known for their antimicrobial activity. Whereas, the antimicrobial activity of A. garcini was well documented in the research work carried out by [6, 7]. Moreover, the in vitro anti-tumor and anti- HIV activities of the above mentioned sesquiterpene lactones have been also reported [8]. Techniques commonly employed for extraction of essential oils is hydro distillation scientific method [9]. The aim of this research study is to identify compounds obtained from Anvillea garcini through hydro distillation method and review of antimicrobial and antioxidant properties were herein discussed. MATERIALS AND METHODS Plant Material All parts of the plant of A.garcini in May 2013 Hormozgan provinces at an altitude of 880 m (Bandar Abbas) in southern Iran were collected. Samples of the plant Medicinal Herbs Research Center was identified in Bandar Abbas. All parts of the plant were kept dried in shade at room temperature (25ْc). Isolation Procedure 70 g of air-dried plant material from each population were cut in small pieces, and the essential oils were obtained by hydro distillation in 350 ml H2O for 3 h [10]. In a Clevenger apparatus [11]. Thus, the oil obtained was dried over anhy-drous sodium sulphate and kept at 4–5 °C in refrigerator till analysis [12]. Available Online at www.ijprbs.com

216



GC and GC Mass The essential oil was analyzed by GC coupled to mass spectrometry (GC-MS) (gas chromatograph model 8500 and mass spectrometer model 5973, Hewlett- Packard, Palo Alto, CA, USA) using a DB-1 fused- silica capillary column (30m × 0.25mm i.d., film thickness 0.25 µm)[13]. The oven temperature was programmed to increase from 60 to 280 ◦C at a rate of 4 ◦C for 1minute and finaly held isothermal for 4 minutes. The carrier gas was helium introduced at a rate of 2mL min−1. Diluted samples (1.0 µL, 1/100 in ether) were injected manually and the split ratio was adjusted to 40:1. GC/MS analyses were performed using a Thermo FinniganTRACEGC (Waltham,MA,USA) coupled with a TRACE MS plus (EI 70 eV) of the same company[14]. Compounds were identified by comparison of their retention indices (RI), determined according to the Kovats method using n-alkanes as standards [15]. The components of the oil were identified by comparison of their mass spectra with those of a computer library or with authentic compounds and confirmed by comparison of their retention indices, there with those of authentic compounds or with data published in the literature [16]. Micro-organisms One gram-positive bacteria Staphylococcus aureus (ATCC 1337) and gram negative bacteria Klebseilla oxytoca (ATCC 1402), three fungal Aspergils niger (ATCC 5154), Fusarium solani (ATCC 5284), Alternaria alternaria (ATCC 5224) were used. All the microorganisms were prepared at Institute of Biological Research, Islamic Azad University, Firozabad IRAN. Antimicrobial Tests The antibacterial activity was evaluated by determining the minimum inhibitory concentration (MIC) using the broth dilution method [17]. Antibacterial evaluation of the essential oil was performed in 96-well plates. Samples of cultures grown overnight (108 cfu/ml) were incubated with oil dissolved in ethanol 99% (at a concentration of 1, 10, 100 and 500 mg/ml) for 24 h at 37 _◦C. All inhibition assays were carried out in triplicate [18]. Gentamicin was used as the reference standard. MIC value was determined, as the lowest concentration of the sample that not permits any visible growth of the tested microorganism after incubation [19]. MIC agar dilution assay The lowest concentration of the compounds that prevented visible growth was considered to be the minimal inhibitory concentration (MIC). In antifungal activity evaluation, appropriate amounts of the natural compounds or essential oil of A. garcini were added aseptically to sterile molted sabouraud dextrose agar (SDA) medium containing 20 (0.5%, v/v) to produce the concentration range of 8-512 µg/mL. The resulting SDA agar solutions were immediately mixed and poured into petri plates. The plates were spot inoculated with 5 µL (104 spore/mL) of Available Online at www.ijprbs.com

217



fungus isolate at the end of incubation period; the plates were evaluated for the presence or absence of growth. The antibacterial activity was carried out similarly through the aforementioned protocol. The only difference is using 5 µL of suspension containing 108 CFU/mL of bacteria instead of fungus isolate. The MIC was defined as the lowest concentration of the oil to inhibit the growth microorganisms [20]. DPPH free radical-scavenging assay This spectrophotometric assay uses the stable DPPH radical as the reagent [21]. The ability of oil to scavenge free radicals of A.garcini essential oil was assayed with the use of a synthetic free radical compound 1, 1- diphenyl-2-picrylhydrazyl (DPPH), followed according to the method employed by Bersuder et al [22]. Further, determination of antioxidant activity was performed in triplicate by a spectrophotometric method. The technique consisted of incubation for 10 min in 500 μL of an ethanol solution of DPPH 0.1 mmol/L with 500 μL of solutions containing increasing concentrations of A.garcini essential oil (12/5-1600 µg/mL) in ethanol. We proceeded similarly to the preparation of the solution called “control” but substituting 500 μL of sample to500-μL ethanol solvent. To a solution called “white”, ethanol was used and the oil in the same concentration range studied [23]. The mixture was then shaken vigorously and left to stand at room temperature for 30 min in the dark [24]. The absorbance was measured at 515 nm using a spectrophotometer as suggested in [25] research work. Moreover, free radical scavenging activity was carried out of using the following the formula; (%I) = [(A blank -A sample) / A blank] × 100 Wherein a blank is the absorbance of the control reaction (containing all reagents apart from the test compound) and a sample is the absorbance of the test compound [24]. The IC 50 value, which represented the concentrations of the essential oil and extracts that caused 50% inhibition, was determined by linear regression analysis from the obtained RSC values [26]. Results and Discussion The GC and GC–MS analyses of the essential oil were obtained from the all parts of A. garcini and allowed the identification of 25 compounds (Table 1), which were representing 97.44% of the oil [27]. The main constituents of this oil were myristicin (58/79%), Bicyclo[5.3.0]decane, 2methylene-5-(1-methylvinyl)-8-methyl- (7/71%) , 5-Methyl-4-nonene (3/46%) . (E)-Ocimene (3/39%). The chemical composition the essential oil of A.garcini Poly phenolic compounds was clearly recorded through as suggested method.


218



Table 1: Chemical composition of A.garcini essential oils obtained by hydro distillation Sl. no. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

Compounds 4-Decene, (E)(-)-α-Pinene 4-Isopropylheptane 1,2- Dimethyl cyclopentane verbenene 5-Methyl-4-nonene Decene (E)-5-Decene Z-Ocimene (E)-Ocimene α-Dodecene 2-Tetradecene, (E).Methyleugenol α-Guaiene Isohomogenol Guaia-1(10),11-diene myristicin ɲ-Selinene Cedrane-8,13-diol β-Eudesmol ç-Gurjunene Hexahydrofarnesyl acetone Hydrofol Morflex 190 Bicyclo[5.3.0]decane, 2-methylene-5(1-methylvinyl)-8-methylNote: KI=….., %HD=….

KI 929 935 945 949 968 981 990 993 1036 1047 1191 1391 1405 1442 1499 1510 1526 1602 1618 1657 1660 1845 1958 1965 2131

%HD 2.24 0.91 0.58 0.58 2.21 3.46 1.41 1.45 1.69 3.39 1.28 0.70 1.73 0.99 1.13 0.97 58.79 0.75 0.72 0.98 1.21 1.05 0.71 0.74 7.71

Figure-1: The constituent KI in different share of compounds obtained through hydro distilation method in A. garcini Available Online at www.ijprbs.com

219



Figure-2: The percentage (%) of HD for different share of compounds obtained through hydrodistilation method in A. garcini Antimicrobial activity The antimicrobial activity of A. garcini was also observed in this research work and according to few investigators revealed that, the ability of essential oil to disrupt the permeability barrier of cell membrane structures and the accompanying loss of chemiosmotic control are the mostly likely reasons for its lethal action [28]. This antimicrobial activity against bacteria and fungi has also been demonstrated in essential oils extracted from A.garcini seed [29]. Antimicrobial and antifungal activity as evidenced by their zones of inhibition shown (table 2). Table 2: Antimicrobial activity of essential oils of A.garcini (measured in unit mg/ml) Tested organism Essential A.garcini

S.aureus

oil 256

K.oxytoca

A.alternaria

F.solani

A.niger

8

32

32

64


220



Figure-3: The share-wise antimicrobial activity (measured in unit mg/ml) in different organisms of A. garcini-essential oil Essential Oil of A.garcini resistant kellebsila Oxytoca was showed gram-negative bacteria and two fungus Alternaria alternaria, Fusarium solani respectively. Further, this activity may be attributed to a synergistic effect, due to the presence in the oil of phenols and some monoterpenes which were possess antimicrobial activity [30-31-32]. Antioxidant activity Results obtained antioxidant activity (DPPH Test) essential oils Shown (Tabel 3). The hydrogen atoms or electron-donating ability of the corresponding essential oil was determined from the bleaching of purple-coloured methanol solution of DPPH [33]. Free radical scavenging activity for essential oil A.garcini was values (IC50=0.0209 ± 0.0028). Radical activity due to essential oil Poly phenolic compounds Such as chemical composition myristicin was. Tabel 3: Antioxidant activity of essential oil A.garcini Plant name

DPPH IC50(mg/ml)

A.garcini

0.0209 ± 0.0028

ACKNOWLEDGEMENTS The first author wishes to express sincere thanks to the Faculty and Chairman (Head of the Department), Department of Chemistry, Firoozabad branch, Islamic Azad University, Firoozabad, Fars, IRAN, for extending the laboratory facilities to carry out the research work.


221



REFERENCES 1. Diallo, D., Hveem, B., Mahmoud, M. A., Betge, G., Paulsen, B. S. & Maiga, A. (1999). An ethnobotanical survey of herbal drugs of Gourma district, Mali. Pharmaceutical Biology, 37, 8091. 2. Farjam. Mh, M. Khalili, A. Rustayian, K. Javidnia and S. Izadi,.(2011). Biological activity of the n-butanolic extract of Stachys pilifera ,African., Journal of Microbiology Research, 5115-5119, 30. 3. Denys J. Charles and James E. Simon.(1990). Comparison of Extraction Methods for the Rapid Determination of Essential Oil Content and Composition of Basil. Journal of American Society Horticulture Sciences. 115(3):458-462. 4. Virendra S. Rana, Jitendra P. Juyal, M. Amparo Blazquez and Surendra H. Bodakhe . (2003).Essential oil composition of Artemisia parviflora aerial parts, Flavour Fragr. J. 18: 342– 344. 5. Rustaiyan A.,T. Naji-Hosseinzadeh , M. Behnam & M. Tabatabaei Anaraki. (2011). Composition and Antimicrobial Activity of the Essential Oil from Leaves and Flowers of Anvillea garcini (Burm.) DC, Journal of Essential Oil Research. 6. Niño J, Narváez DM, Mosquera OM, Correa YM.(2006). Antibacterial, antifungal and cytotoxic activities of eight Asteraceae and two Rubiaceae plants from Colombian biodiversity. Braz J Microbiol. 566-70. 7. Paulo S.(2006). Antibacterial, antifungal and cytotoxic activities of height Asteraceae and two Rubiaceae plants from Columbian biodiversity. Brazilian J Microbial. 321-35. 8. Abdel-Sattar, E. A. Galal and G.S. Mossa, (1996). Antitumor Germacranolides from Anvillea garcini. J. Nat. Prod., 59, 403-405 . 9. Asta. (1968). Official analytical methods of the American Spice Trade Association. Asta, Englewood Cliffs, N.J. p. 8-11. 10. Hellenic Pharmacopoeia, ed IV. vol 1 Atlantis-Pechlyvanides: Athens, 1989; 191. 11. Guido C. Galletti and Maria Teresa Russo, (1996). Essential Oil Composition of Leaves and Berries of Vitex agnus-castus L. from Calabria, Southern Italy, RAPID Communications In Mass Spectrometry,. 10, 1345-1350.


222



12. Raina, V. K. Srivastava, S. K. Aggarwal, Ł K. K. Syamasundar, K. V. and Sushil Kumar. (2001). Essential oil composition of Syzygium aromaticum leaf from Little Andaman, India, Flavour Fragr. J. 334–336. 13. Khalfi.O, N. Sahraoui, F. Bentahar and Boutekedjiret, Ch. (2008). Chemical composition and insecticidal properties of Origanum glandulosum (Desf.) essential oil from Algeria, J Sci Food Agric. 1562–1566. 14. Yang Yang, Run-Wei Yan, Xiao-Qiang Cai, Zhong-Liang Zheng and Guo-Lin Zou. (2008).Chemical composition and antimicrobial activity of the essential oil of Amomum tsao-ko, J Sci Food Agric 2111–2116 . 15. Jennings W and Shibamoto T. (1980). Qualitative Analysis of Flavour and Fragrance Volatiles by Glass Capillary Gas Chromatography. Academic Press, New York. 16. Adams, R. P. (1995). Identification of essential oil components by gas chromatography/mass spectroscopy. Carol Stream, USA: Allured Publishing Corp. 17. Koneman, E.W., Allen, S.D., Janda, W.M., Schreckenberg, P.C. and Winn W.C. (1997). Color Atlas and Textbook of Diagnostic Microbiology. Lippincott-Raven pub, Philadelphia, PA: pp. 7845. 18. Ben Sassi .A, F. Harzallah-Skhiri, I. Chraief, N. Bourgougnon, M. Hammami, M. Aouni. (2008). Chemical composition and antimicrobial activities of the essential oil of (Tunisian) Chrysanthemum trifurcatum (Desf.) Batt. and Trab. Flowerheads, C. R. Chimie. 324e330. 19. Juarez Belaunde.Aj , J G. Sandoval , L. De Martino, F. Senatore, V. De Feo.(2007). Chemical Composition and Antibacterial Activity of Senecio nutans Essential Oil, Jeobp. 332 – 338. 20. Ezzatzadeh.E, M H. Farjam, A. Rustaiyan. (2012). Comparative evaluation of antioxidant and antimicrobial activity of crude extract and secondary metabolites isolated from Artemisia kulbadica, Asian Pacific Journal of Tropical Disease .431-434. 21. Burits, M., Bucar, F. (2000). Antioxidant activity of Nigella sativa essential oil. Phytother Res., 14: 323-328 22. Bersuder, P., Hole, M. and Smith, G. (1998). Antioxidants from a heated histidineglucose model system I: Investigation of the antioxidant role of histidine and isolation of antioxidants by high performance liquid chromatography. J. Am. Oil Chem. Soc. 75: 181-187. 23. Adriana B. Vanin & Tainara Orlando & Suelen P. Piazza Bruna M. S. Puton & Rogério L. Cansian & Debora Oliveira & Natalia Paroul. (2014). Antimicrobial and Antioxidant Activities of


223



Clov Essential Oil and Eugenyl Acetate Produced by Enzymatic Esterification, Appl Biochem Biotechnol . 174:1286–1298 24. Sebnem Yilmaz .E, M. Timur, and B. Aslim. (2013). Antimicrobial, Antioxidant Activity of the Essential Oil of Bay Laurel from Hatay, Turkey, TEOP. 108 – 116. 25. Farhoudi.R. (2013). Chemical Constituents and Antioxidant Properties of Matricaria recutita and Chamaemelum nobile Essential Oil Growing Wild in the South West of Iran, Journal of Essential Oil Bearing Plants. 16 4, p531-p537, 7p. 26. Salehi. P, A. Sonboli, F. Eftekhar, S. Nejad-Ebrahimi, and Yousefzadi, M. (2005). Essential Oil Composition, Antibacterial and Antioxidant Activity of the Oil and Various Extracts of Ziziphoraclinopodioides subsp. rigida (BOISS.) RECH. f. from Iran, Biol. Pharm. Bull. 28(10) 1892—1896. 27. Senatore. F, F. Napolitano, N. Apostolides Arnold, M Bruno and W. Herz. (2005). Composition and antimicrobial activity of the essential oil of Achillea falcata L. (Asteraceae), Flavour Fragr. J. 20: 291–294. 28. Cox, S. D., Mann, C. M., Markham, J. L., Bell, H. C., Gustafson, J. E., Warmington, J. R., et al. (2000). Mode of antimicrobial action of essential oil of Melaleuca alternifolia tea tree oil. Journal of applied microbiology, 88(1), 170–175. 29. Lo Cantore, P., Iacobellis, N., De Marco, A., Capasso, F., & Senatore, F. (2004). Antibacterial activity of Coriandrum sativum L. and Foeniculum vulgare miller var. vulgare (miller) essential oils. Journal of Agricultural and Food Chemistry, 52, 7862–7866. 30. Aligiannis N, Kalpoutzakis E, Kyriakopoulou I, Mitaku S, Chin IB.(2004). Flavour Fragr. J; 19: 320–324. 31. Kim J, Marshall MR, Wei C.(1995). Antibacterial activity of some essential oil components against five foodborne pathogens .J. Agric. Food Chem. 43: 2839–2845. 32. Griffin SG, Wyllie SG, Markham JL, Leach DN. (1999). The role of structure and molecular properties of terpenoids in determining their antimicrobial activity Flavour Fragr. J; 14: 322–332 33. Hatano T, Kagawa H, Yasahara HT, Okuda T. (1988). Two new flavonoids and other constituents in licorice root: their relative astringency and radical scavenging effects. Chem Pharm Bull (Tokyo). 36(6):2090–2097.


224