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Total Phenolic Content, Flavonoid Concentration, Antioxidant and Antimicrobial Activity of Methanol Extracts from Three Seseli L

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Total Phenolic Content, Flavonoid Concentration, Antioxidant and Antimicrobial Activity of Methanol Extracts from Three Seseli L Cent. Eur. J. Biol. • 7(6) • 2012 • 1116-1122 DOI: 10.2478/s11535-012-0094-4 Central European Journal of Biology Total phenolic content, flavonoid concentration, antioxidant and antimicrobial activity of methanol extracts from three Seseli L. taxa Research Article Jelena S. Matejić1, Ana M. Džamić2, Tatjana Mihajilov-Krstev3, Vladimir N. Ranđelović3, Zoran Đ. Krivošej4, Petar D. Marin2,* 1University of Niš, Faculty of Medicine, 18000 Niš, Serbia 2University of Belgrade, Faculty of Biology, Institute of Botany and Botanical Garden “Jevremovac”, 11000 Belgrade, Serbia 3University of Niš, Faculty of Science and Mathematics, 18000 Niš, Serbia 4University of Pristina, Department of Biology, Faculty of Natural Sciences, 38220 Kosovska Mitrovica, Serbia Received 19 April 2012; Accepted 14 August 2012 Abstract: Thepresentstudydescribesthetotalphenoliccontent,concentrationsofflavonoidsandin vitroantioxidantandantimicrobialactivityof methanol extracts from Seseli pallasii Besser, S. libanotis (L.) Koch ssp. libanotis and S. libanotis (L.) Koch ssp. intermedium (Rupr.) P. W. Ball, growing wild in Serbia. The total phenolic content in the extracts was determined using Folin-Ciocalteu reagent and their amounts rangedbetween84.04to87.52mgGA(gallicacid)/g.Theconcentrationsofflavonoidsintheextractsvariedfrom4.75to19.37mg Qu(quercetin)/g.AntioxidantactivitywasanalyzedusingDPPHreagent.Antioxidantactivityrangedfrom0.46to4.63IC50 (mg/ml) and from1.98to2.19mgVitC(vitaminC)/gwhentestedwiththeDPPHandABTSreagents,respectively,usingBHAandVitCascontrols. Theantimicrobialactivityoftheextractswasinvestigatedusingamicro-welldilutionassayforthemostcommonhumangastrointestinal pathogenic bacterial strains: Escherichia coli ATCC25922,Pseudomonas aeruginosaATCC9027,Salmonella enteritidisATCC13076, Bacillus cereusATCC10876,Listeria monocytogenesATCC15313,Staphylococcus aureusATCC25923andCandida albicans ATCC 10231.ThisfindingsuggeststhatSeseli species may be considered as a natural source of antioxidants and antimicrobial agents. Keywords: Seseli pallasii • S. libanotis • Apiaceae • Antioxidant and antimicrobial activity • DPPH, ABTS • Phenols • Flavonoids ©VersitaSp.zo.o. 1. Introduction S. libanotis (L.) Koch ssp. libanotis has lower leaves that are 2- to 3-pinnate, lobes linear, oblong or The genus Seseli belongs to the family Apiaceae lanceolate, often falcate, acute. This taxon is distributed (=Umbelliferae) and contributes 10 species to the flora in western, central and parts of south Europe, extending of Serbia [1]. Seseli pallasii Besser. (incl. S. varium northwards to southern Sweden [2]. Trev.) - moon carrot, is a glabrous biennial or perennial Seseli libanotis (L.) Koch ssp. intermedium (Rupr.) plant that reaches a height of 30-120 cm. Its leaves vary P. W. Ball is a plant with lower leaves 1- to 2-pinnate, from 2- to 4-pinnate linear to almost filiform. Its petals lobes ovate, coarsely toothed or pinnatifid, obtuse or are white and glabrous, and its fruit is ellipsoid or oblong, subobtuse. This taxon is distributed in eastern and glabrous or slightly tuberculate-verrucose. This taxon is central Europe [2]. distributed in the north of Italy, the Czech Republic and A screening of S. pallasii extracts for chronic Slovakia eastwards to central Ukraine [2]. toxicity and larval inhibition on Spodoptera littoralis * E-mail: [email protected] 1116 J.S. Matejić et al. larval growth was recently reported. Lethal doses does not occur with the ABTS assay, especially when and the effect of LD50 doses on growth inhibition and the absorbance is measured at 734 nm [11]. antifeedant were estimated in order to determine the In this study, the antioxidant and antimicrobial potency of the selected extracts based on the mortality activity of Seseli pallasii, S. libanostis ssp. libanotis, and results [3]. S. libanotis subsp. intermedium methanol extracts is In the Eastern part of Turkey, S. libanotis is a reported. As far as we know this is the first report of the common plant used both as a cheese preservative antioxidant activity of these taxa. and to provide aroma. Besides S. libanotis, 25 different species can be used to make herb-flavoured cheese. These herbs, e.g. Allium spp., Thymus spp., Falcaria 2. Experimental Procedures spp. etc., can be added singly or as a mixture to cheese [4]. Leaves of S. libanotis (kelemkesir or kelemenkesir 2.1 Chemicals in Turkish) are consumed as a vegetable in eastern Organic solvents were purchased from “Zorka Turkey [5]. The methanol extract of this plant had a pharma” Sabac, Serbia. Gallic acid, 3-tert-butyl- broad-spectrum antibacterial activity against Bacillus 4-hydroxyanisole (BHA) and 2,2-dyphenyl-1- cereus, B. dipsauri, B. lentimorbus, B. sphaericus, picrylhydrazyl (DPPH) were obtained from Sigma B. subtilis, Corynebacterium ammoniagenes, Kocuria Chemicals Co., St Louis, MO, USA. Folin-Ciocalteu rosea, Neisseria subflava and Micrococcus lylae, which phenol reagent was purchased from Merck, is of interest in relation to the prevention of microbial Darmstadl, Germany. Sodium carbonate anhydrous contamination in foods [6]. (Na2CO3), potassium acetate (C2H3KO2), potassium Recently, the results of essential oil analyses and peroxidisulphate (K2O8S2) and L(+)- Ascorbic acid antimicrobial effects of different Seseli species were (Vitamin C) were purchased from AnalaR Normapur, published. The essential oil from the aerial parts of VWR, Geldenaaksebaan, Leuven Belgium. Aluminium S. annuum, wild-growing in Serbia, was characterized nitrate nonahydrate (Al(NO3)3x9H2O) was purchased as containing germacrene-D, sabinene, Z-β-ocimene from Fluka Chemie AG, Buchs, Switzerland. ABTS and limonene. This oil showed antifungal activity against and quercetin hydrate were obtained from TCI Europe fifteen fungi with MICs between 12.5 to 50 μl/ml 7[ ]. NV, Boerenveldsweg, Belgium. All other solvents and Also, essential oil from aerial parts of S. globiferum Vis. chemicals were of analytical grade. which contains sabinene, α-pinene and β-phellandrene showed activity against Pseudomonas aeruginosa, 2.2 Plant material Micrococcus flavus, Lysteria monocytigenes and Aerial plant parts of S. pallasii were collected in July 2003 Escherichia coli, and all investigated micromycetes from Soko Grad (Soko Banja) and parts of S. libanotis [8]. The essential oil isolated from S. montanum ssp. ssp. libanotis were collected in July 2003 from the tommasinii (Reichenb. fil.) Arcangeli was also analyzed Kopaonik mountains. Aerial plant parts S. libanotis and found to possess moderate to strong antimicrobial ssp. intermedium were collected in July 2009 and activity, which can be explained by presence of the fruits were collected in September 2009 from the Stara β-pinene, germacrene-D, sabinene, α-pinene and planina mountains. Voucher specimens for S. pallasii, limonene [9]. The essential oil of S. rigidum which S. libanotis ssp. libanotis (LM 60313), S. libanotis ssp. contains α-pinene, camphene, β-pinene, and limonene intermedium (SL 16433), have been deposited at the was also found to have antimicrobial and potential Herbarium of the Institute of Botany and Botanical antioxidant activity [10]. Garden “Jevremovac”, Faculty of Biology, University of Several methods are available to evaluate Belgrade. antioxidant activities of natural compounds in foods or biological systems. Two methods commonly used 2.3 Preparation of plant extracts in antioxidant activity assays are the 2,2-dyphenyl- Plant material was air dried in the dark and ground to 1-picrylhydrazyl (DPPH) and 2,2’-azinobis(3- a powder. The aerial plant parts (10 g) were powdered ethylbenzthiazoline-6-sulfonic acid) (ABTS) tests. ABTS and extracted with 100 ml methanol. The mixture was is soluble in both aqueous and organic solvents, and exposed to ultrasound for 30 min and after 24 h standing it reacts relatively rapidly compared to DPPH, which in the dark was filtered. The methanol solvent was normally takes several hours for the reaction to be removed by evaporation under reduced pressure, at a completed. Color interference of the DPPH assay with maximum temperature of 40ºC. After evaporation of the samples that contain anthocyanins leads to under- solvent, the crude extract was subjected to subsequent estimation of antioxidant activity. However, this problem analysis. 1117 Total phenolic content, flavonoid concentration, antioxidant and antimicrobial activity of methanol extracts from three Seseli L. taxa 2.4 Determination of total phenolic content Sample concentrations for each compound and The total phenolic content of extracts was determined standard antioxidant, which decrease absorption of spectrophotometrically by the Folin-Ciocalteu method DPPH solution to 50% (IC50), were obtained from according to the procedure reported by Singleton et al. the absorption of curves DPPH solution at 517 nm. with some modifications [12]. Briefly, 300 µl of methanol Calculations were performed with Origin 7.0 software. extracts solution and 1500 µl of 1:10 Folin-Ciocalteau reagent were mixed and after 6 minutes in the dark 2.7 Evaluation of ABTS radical scavenging 1200 µl of sodium carbonate (7.5%) was added. After activity 2 h of incubation in the dark at room temperature, the The ABTS radical-scavenging activity, was determined absorbance at 740 nm was measured (Shimadzu, following the method of Miller and Rice-Evans with UV-Visible PC 1650 spectrophotometer). The total some modifications [15]. The ABTS·+ solution was phenolic concentration was calculated from a gallic prepared by mixing 19.2 mg of ABTS with
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