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In Vitro Antioxidant and Antimicrobial Activities of Aerial Parts of Algerian Jurinea Humilis DC (Asteraceae)

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In Vitro Antioxidant and Antimicrobial Activities of Aerial Parts of Algerian Jurinea Humilis DC (Asteraceae) Ayad et al Tropical Journal of Pharmaceutical Research December 2017; 16 (12): 2903-2909 ISSN: 1596-5996 (print); 1596-9827 (electronic) © Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, 300001 Nigeria. Available online at http://www.tjpr.org http://dx.doi.org/10.4314/tjpr.v16i12.14 Original Research Article In vitro antioxidant and antimicrobial activities of aerial parts of Algerian Jurinea humilis DC (Asteraceae) Radia Ayad1*, Yavuz S Cakmak2, Meltem A Ozusaglam2, Kamel Medjroubi1, Salah Akkal1 1Varenbiomol, Department of Chemistry, University Mentouri Constantine 1, 25000 Constantine, Algeria, 2Department of Biotechnology and Molecular Biology, Faculty of Science and Arts, Aksaray University, Aksaray, Turkey *For correspondence: Email: [email protected]; Tel: +21331811102 Sent for review: 25 September 2017 Revised accepted: 22 November 2017 Abstract Purpose: To assess the polyphenolic composition of various solvent extracts of Jurinea humilis DC. as well as their in vitro biological effects. Methods: The crude extracts of the aerial parts of Jurinea humilis were obtained by maceration method with dichloromethane (DCM), methanol (MeOH) and ethyl acetate (EtoAC), separately. Folin-Ciocalteu and aluminum chloride procedures were used to quantify the total phenolic and flavonoid contents, respectively, while antioxidant properties were determined by two methods: phosphomolybdenum and DPPH assays. Antimicrobial activity was also evaluated by disc diffusion method. Results: The ethyl acetate extract exhibited the highest amount of total phenolic content (TPC, 169.14 ± 7.22 mg gallic acid equivalent/g sample) and total flavoniod contents (TFC, 104.91±0.22 mg rutin equivalent/g sample). The extract (EtoAC) also showed the strongest antioxidant activity of the three extracts, especially DPPH assay (IC50 = 0.16 ± 0.00 mg/mL), as well as demonstrated the highest activity against the pathogenic strains tested. Conclusion: The results suggest that J. humilis DC. is rich in phenolic and flavonoid contents which may be responsible for good antioxidant and antimicrobial properties. Thus, J. humilis may be a promising source of new antioxidant agents and pharmaceuticals. Keywords: Jurinea humilis, Phenolic, Flavonoid, Antioxidant, Antimicrobial This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. Tropical Journal of Pharmaceutical Research is indexed by Science Citation Index (SciSearch), Scopus, International Pharmaceutical Abstract, Chemical Abstracts, Embase, Index Copernicus, EBSCO, African Index Medicus, JournalSeek, Journal Citation Reports/Science Edition, Directory of Open Access Journals (DOAJ), African Journal Online, Bioline International, Open-J-Gate and Pharmacy Abstracts INTRODUCTION the genus Jurinea is represented by a single species, Jurinea humilis DC. [3]. Jurinea Cass. (Compositae, tribe Cynareae, subtribe Carduineae) is the largest genus in the Although a significant number of plants belonging family Asteraceae, including about 200 species to Jurinea species contain very useful chemical that are found in diverse regions, especially constituents, very few papers have been Central Asia, as well as Iran, Turkey and the published on their phytochemical compositions. Mediterranean basin [1,2]. In the flora of Algeria, These chemical studies have revealed the ----------------------------------------------------------------------------------------------------------------------------------------------------- © 2017 The authors. This work is licensed under the Creative Commons Attribution 4.0 International License 2903 Ayad et al presence of several constituents, chief among water, and incubated for 2 h in the dark at room them, were sesquiterpene lactones and temperature. The absorbance value was triterpenes [4]. Recently, phytochemical measured at 765 nm using a UV/Visible investigation of J. dolomiaea roots demonstrated spectrophotometer (Thermo Electron Corporation the presence of caffeic acid, apigenin, catechin evolution 100). The total phenolic content was and rutin [5]. Jurinea species have been known presented as gallic acid equivalent per gram of for their numerous activities like antimicrobial, dry extract (GAE mg/g extract). antioxidant, anticholinesterase, antilipid peroxidation, anti-toxic and antileishmanial Total flavonoid content of extract samples was activity [4]. With regard to their quantitative measured also [9]. Briefly, a mixture of equal composition (total phenolic and flavonoid volumes (1 mL) of 2 mg/mL extract and 2 % contents), there are only three previous reports methanolic aluminium chloride (AlCl3) solution on J. dolomiaea and J.consanguinea [5-7]. was prepared and incubated for 10 min. After, the absorbance was read at 415 nm. Methanol The objectives of this study were to evaluate the was used as blank. Results were presented as chemical composition (total phenolic and rutin equivalent per gram of dry extract (mg RE/g flavonoid contents) of various extracts of the of extract). aerial parts of J. humilis DC. as well as their in vitro antioxidant potentials and antimicrobial Total antioxidant capacity activity against different strains of microorganisms. Total antioxidant capacity of samples was determined according to phosphomolybdate To best of our knowledge, no previous chemical method of Saeed et al. with slight modification composition and biological activities have been [10].In brief, 3 mL of reagent solution (6 M reported for Jurinea humulis DC. sulfuric acid, 28 mM sodium phosphate, and 4 mM ammonium molybdate) were added to 0.3 EXPERIMENTAL mL of each extract (2 mg/mL). The absorbance was measured against blank at 695 nm using a Plant material UV/visible spectrophotometer (Thermo Electron Corporation evolution 100), after incubation for J. humilis DC. were collected in May 2014 from 90 min at 95 °C. Antioxidant capacity of the Boussaada (Algeria), and identified by Professor extracts was expressed as ascorbic acid (mg Hossine Laouar, Laboratory of Natural and AE/g extract) equivalent. Biological Resources Valorization, Department of Biology and Plant Ecology. A voucher specimen Free radical scavenging capacity (DPPH, 2, 2- was archived in the herbarium of the same diphenyl-1-picrylhydrazyl) laboratory of University Ferhat Abbes, Setif 1 (Voucher no. JH-LH-2014). The free radical scavenging activity of extracts was investigated by the method of Kirby and Preparation of plant extracts Schmidt with a partial modification [11]. Briefly, each 0.5 mL of diluted solutions (0.2 - 1 mg/mL) The collected plant material was cleaned and of extracts in methanol was mixed with 3 mL of −5 dried. Each 50 g of dried plant was separately DPPH methanolic solution (6.10 M). The extracted by marceration using 200 mL of the mixture was allowed to react in the dark for 30 following solvents: dichloromethane (DCM), ethyl min at room temperature. acetate (EtoAC) or methanol (MeOH) at room temperature for three days. The filtrates were Absorbance was measured at 517 nm using a evaporated to dryness under vacuum using a UV/visible spectrophotometer (Thermo Electron rotary evaporator at 45 °C. Corporation evolution 100). The control was prepared as above without any extract. Inhibition Determination of antioxidant activity (H) was determined as in Eq 1. Total phenolic and flavonoid contents H(%) = (A0 − A1)/A0}100 ………. (1) Total phenolic content was quantified using the where A0 = absorbance of the control, A1 = Folin–Ciocalteu method [8].The appropriate sample/absorbance of standard), BHT was used dilutions (2mg/mL) of 0.2 mL of plant extracts as standard. The results are expressed as were oxidized with 1 mL Folin–Ciocalteu reagent. median inhibitory concentration (IC50). After, 2 mL of 7.5% Na2CO3 were added. The mixture was then stirred in 7 mL of distilled 2904 Ayad et al Determination of antimicrobial activity extracts two fold dilutions were followed [13,14]. The strains that showed an inhibition zone in our Microbial strains assay were subjected to determine their MBC and MFC values. Therefore, each of extracts: the The antimicrobial activity of all extracts was dichloromethane (DCM), ethyl acetate (EtoAC) tested against a total of 13 microorganisms. and methanol (MeOH) were dissolved in test These included fish pathogenic strains, one tubes, at the initial concentration of 60.00 mg/mL, Gram positive bacteria: Yersinia ruckeri, and two and then varying dilutions were obtained even to Gram negative bacteria: Lactococcus garvieae, achieve final concentration of 0.94 mg/mL. At Vibrio anguillarum (A4 strains, obtained from first, the microorganisms were inoculated during different companies), added to clinical and food- 12h and the resulting cultures suspensions were borne pathogenic microorganisms. Gram positive adjusted to 0.5 McFarland. A volume of 2.5 μL of bacteria: Bacillus cereus (RSKK 86), the tested microorganism was introduced to each Micrococcus luteus (NRRL B-4375), tube and further completed by using 100 μL of Staphylococcus aureus (ATCC 25923), Listeria the respective medium. monocytogenes (ATCC 7644) and Gram negative bacteria: Yersinia enterocolitica (NCTC Positive
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