GC/MS Evaluation and in Vitro Antioxidant Activity of Essential Oil and Solvent Extracts of an Endemic Plant Used As Folk Remedy in Turkey: Phlomis Bourgaei Boiss
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Hindawi Publishing Corporation Evidence-Based Complementary and Alternative Medicine Volume 2013, Article ID 293080, 7 pages http://dx.doi.org/10.1155/2013/293080 Research Article GC/MS Evaluation and In Vitro Antioxidant Activity of Essential Oil and Solvent Extracts of an Endemic Plant Used as Folk Remedy in Turkey: Phlomis bourgaei Boiss. Cengiz Sarikurkcu,1 M. Sabih Ozer,2 Ahmet Cakir,3 Mustafa Eskici,2 and Ebru Mete4 1 DepartmentofGastronomyandCulinaryArts,FacultyofTourism,UniversityofNecmettinErbakan,Meram,Konya42100,Turkey 2 Department of Chemistry, Faculty of Science and Literature, Celal Bayar University, Muradiye, Manisa 45030, Turkey 3 Department of Chemistry, Faculty of Arts and Sciences, Kilis 7 Aralık University, Kilis 79000, Turkey 4 Department of Chemistry, Faculty of Science, Ataturk¨ University, Erzurum 25240, Turkey Correspondence should be addressed to M. Sabih Ozer; [email protected] Received 10 February 2013; Accepted 12 April 2013 AcademicEditor:F.R.F.Nascimento Copyright © 2013 Cengiz Sarikurkcu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. This study was outlined to examine the chemical composition of hydrodistilled essential oil and in vitro antioxidant potentials of the essential oil and different solvent extracts of endemic Phlomis bourgaei Boiss. used as folk remedy in Turkey. The chemical composition of the oil was analyzed by GC and GC-MS, and the predominant components in the oil were found to be - caryophyllene (37.37%), (Z)--farnesene (15.88%), and germacrene D (10.97%). Antioxidant potentials of the solvent extracts and the oil were determined by four testing systems including -carotene/linoleic acid, DPPH, reducing power, and chelating effect. In -carotene/linoleic acid assay, all extracts showed the inhibition of more than 50% at all concentrations. In DPPH, chelating effect, and reducing power test systems, the water extract with 88.68%, 77.45%, and 1.857 (absorbance at 700 nm), respectively, exhibited more excellent activity potential than other extracts (hexane, ethyl acetate and methanol) and the essential oil at 1.0 mg/mL concentration. The amount of the total phenolics and flavonoids was the highest in this extract (139.50 ± 3.98 g gallic acid equivalents (GAEs)/mg extract and 22.71 ± 0.05 g quercetin equivalents (QEs)/mg extract). 1. Introduction that could lead to mutation. All aerobic organisms, includ- ing humans, have antioxidant defenses that protect against Lipid peroxidation is one of the major factors causing dete- oxidative damage [3]. However, these natural antioxidant rioration of foods during the storage and processing. Also, mechanisms can be inefficient, and hence dietary intake of oxidized polyunsaturated fatty acids may induce aging and antioxidant compounds becomes important. Herbal therapy carcinogenesis [1]. Although there are some synthetic antiox- has been widely used to treat a large variety of various ail- idant compounds, such as butylated hydroxytoluene (BHT) ments and symptoms, and there is a recently growing interest and butylated hydroxyanisole (BHA), which are commonly on the herbal therapy in the world. It has also been believed used in processed foods, it has been reported that these that medicinal plants are an important source of alternative compounds can have some side effects [2]. Therefore, many chemical reagents with potential therapeutic effects4 [ , 5]. researchers have focused their attention on natural antioxi- One of the potential uses of plant-derived compounds is dants. Reactive oxygen species (ROS) and reactive nitrogen to be used as antioxidant agents [3, 6, 7]. It is well known species (RNS) are produced in cells by different means [3]. that antioxidants may be useful in protecting from cancer Exogenous sources of free radicals include tobacco smoke, and other mutation-related diseases. The hypothesis is that ionizing radiation, certain pollutants, organic solvents, and the use of antioxidants that scavenge reactive oxygen species pesticides [1]. ROS and RNS may also cause DNA damage (ROS) provides biological resistance to free radicals, retards 2 Evidence-Based Complementary and Alternative Medicine theprocessofaging,anddecreasestheriskofage-associated conditions. The residue was then extracted by boiling water degenerative diseases, such as cancer, cardiovascular diseases, (300 mL). Hexane, ethyl acetate, and methanol in the extracts immune system decline, and brain dysfunction [5]. were removed with a rotary evaporator to obtain the extracts The genus Phlomis as perennial herbs of Lamiaceae family in the yield of 1.44%, 0.75%, and 7.17% (w/w), respectively. The consists of more than 100 species distributed in Europe, Asia, water extract was dried in a freeze drier to obtain an extract and Africa. In Turkish flora, 52 taxa include 6 varieties, 12 in the yield of 11.37% (w/w). natural hybrids, 34 endemic taxa which are distributed [8]. Many species of this genus have usage for medicinal and 2.3. Isolation of the Essential Oil. The air-dried and ground aromatic purposes. The aerial parts of some species including plant material (500 g) was submitted for 5 h to water distilla- Phlomis and Thymus have distinctive tastes and are used for tion using a British-type Clevenger apparatus (ILDAM Ltd., the herbal tea in traditional medicine as stimulants, tonics, Ankara, Turkey) (yield 0.11%). The obtained essential oil was carminative, appetizer, andiuretics [4]. It has been shown dried over anhydrous sodium sulphate and after filtration, ∘ that Phlomis species have exhibited the various biolodgical stored at +4 Cuntiltestedandanalyzed. properties such as in treatment of ulcer and hemorrhoids [7, 9, 10].ItwasalsoreportedthatsomePhlomis species 2.4. Gas Chromatography (GC) Analysis. The analysis of the have anti-inflammatory, antimicrobial [9], antiulcerogenic essential oil was performed using a Thermo Finnigan Trace [10], immunosuppressive [11],andfreeradicalscavenging GC/A1300, (E.I), equipped with a SGE/BPX5 MS capillary [12] activities. Pytochemical investigations of Phlomis species column (30 m × 0.25 mm i.d., 0.25 m). Helium was the were the subject to several studies, and, consequently, iridoid carrier gas, at a flow rate of 1 mL/min. Injector temperature ∘ ∘ and phenyl propanoid glycosides were isolated [7, 13]. was set at 220 C. The programme used was 50–150 Catarate ∘ The Phlomis genus and their essential oils are used in the of 3 C/min, held isothermal for 10 min, and finally raised to ∘ ∘ food and pharmaceutical industries. Essential oils isolated 250 Cat10C/min. Diluted samples (1/100, v/v, in methylene from Phlomis species are used as flavoring for foods and as chloride) of 1.0 Lwereinjectedmanuallyandinthesplitless fragrance in the perfume and cosmetic industry. Therefore, mode. Quantitative data of the oil was obtained from flame there were numerous reports on the chemical analyses of ionization detector (FID) area percentage data. the essential oils of various Phlomis species distributed in different regions of the world [7, 8, 15]. On the other hand, 2.5. Gas Chromatography/Mass Spectrometry (GC/MS) Anal- as far as our literature survey could ascertain, there is only ysis. The analysis of the essential oil was performed with a one report on the chemical composition of the essential oil Thermo Finnigan Trace GC/Trace DSQ/A1300, (E.I Quad- of P. b ourg ae i [16]. Additionally, antioxidant activities of the rapole) equipped with a SGE-BPX5 MS fused silica capillary solvent extracts and the essential oil of this plant have not column (30 m × 0.25 mm i.d., film thickness 0.25 m). For previously been reported. Therefore, data presented here will GC-MS detection, an electron ionization system with ion- be the first record on P. b ourg ae i . ization energy of 70 eV was used. Carrier gas was helium The aim of this study is to determine the potential an- at a flow rate of 1 mL/min. Injector and MS transfer line ∘ ∘ tioxidant and free radical scavenging activities, reducing temperatures were set at 220 Cand290C, respectively. The ∘ ∘ and chelating effects of hexane, ethyl acetate, methanol and oven temperature was programmed from 50 Cto150Cat ∘ water extracts, and essential oil of P. b ourg ae i as well as the 3 C/min, then held isothermal for 10 min, and raised to ∘ ∘ chemical composition of the oil. Additionally, total phenolic 250 Cat10C/min. Diluted samples (1/100, v/v, in methylene and flavonoid contents of the extracts have been determined. chloride) of 1.0 Lwereinjectedmanuallyinthesplitless mode. The identification of individual compounds was based on 2. Materials and Methods comparison of their relative retention times with those of authentic samples on SGE-BPX5 capillary column, and by 2.1. Plant Material. Phlomis bourgaei Boiss. was collected in matching their mass spectra of peaks with those obtained 2008 from Mugla University campus, Mugla, Turkey, at its from authentic samples and/or the Wiley 7N and TRLIB flowering season. Taxonomic identification of the plant mate- libraries spectra and published data [14]. rial was confirmed by the senior taxonomists Dr. Omer Varol and Dr. Olcay Ceylan, in Department of Biology, Mugla 2.6. Antioxidant Activity University. The voucher specimen has been deposited at the HerbariumoftheDepartmentofBiology,MuglaUniversity, 2.6.1. Total Antioxidant Activity by -Carotene-Linoleic Acid Mugla,Turkey (Voucher number