Özek G, Özbek MU, Arslan M. JOTCSA. 2018; 5(3): 1361-1370. RESEARCH ARTICLE Lipid and essential oil constituents of Cota hamzaoglui Özbek & Vural (Asteraceae) Gülmira Özek* , Mehmet Ufuk Özbek2 , Münevver Arslan3 1Anadolu University, Faculty of Pharmacy, Department of Pharmacognosy, 26470, Eskişehir, Turkey 2 Gazi University, Faculty of Science, Department of Biology, 06500, Teknikokullar, Ankara, Turkey. 3 Research Institute for Forest Soil and Ecology, p.b. 61, 26160, Eskişehir, Turkey Abstract: In the present work, lipids and essential oil constituents of endemic Cota hamzaoglui Özbek & Vural were investigated with GC-FID/MS techniques. The fatty acids fraction was isolated with liquid-liquid extraction from the herb with Folch method and then methylated with BF3 reagent. Linolenic, linoleic, oleic, and hexadecanoic acids were found to be the main fatty acids. The unsaturated fatty acids (66.0%) prevailed upon saturated (33.6%) ones. The essential oil was characterized with high percentage of the fatty acids (34.7%), alkanes (14.0%) and aliphatic aldehydes (8.3%). The present study is the first report on chemical composition of Cota hamzaoglui Özbek & Vural lipids and essential oil. Keywords: Cota hamzaoglui Özbek & Vural; essential oil, lipids, GC-FID/MS. Submitted: October 30, 2018. Accepted: December 07, 2018. Cite this: Özek G, Özbek M, Arslan M. Lipid and essential oil constituents of Cota hamzaoglui Özbek & Vural (Asteraceae). JOTCSA. 2018;5(3):1361-70. DOI: http://dx.doi.org/10.18596/jotcsa.476387. *Corresponding author. E-mail: [email protected], Tel: +90-2223350580 Extn.:3716, Fax: +90-2223306809 INTRODUCTION nine of which are endemic (1, 2). Earlier, Cota was recorded as a section in the genus The Asteraceae family contains the largest Anthemis L. in Flora of Turkey (12). Recently, number of described species, approximately the Anthemis section Cota has been accepted 25,000 distributed in over 2,200 genera (1-3). as a generic name, Cota (13, 14). The genus The plants of Asteraceae family have been Cota morphologically resembles Anthemis, found to be the most commonly used families however differs by achenes (2). in the traditional medical treatments in Representatives of the genus Cota have Turkey. Ethnomedicinal aspects of potency of economic importance because of their uses for Asteraceae plants have recently been reported various purposes such as obtaining drug, food by Altundag et al. (4). Importance of the and dye (15). essential oils of the Anthemideae plants has been discussed in recently reported paper by Today there is increasing demand for cheap, Silva (5). Many genera have been approved safe, and scientifically approved botanicals for applying in treatment of a number of from domestic sources. However, there are diseases, Tanacetum (6), Silybum (7), still species have not been investigated for Matricaria (8), Achillea (9), Artemisia (10) and phytochemical and biological potentials. The Anthemis (11). The genus Cota J. Gay is plants of the genus Cota are among less- represented by 63 taxa which are mainly investigated species. In literature, there is distributed in Europe (excluding northern information that the flowers of the genus Cota Europe), North Africa, Caucasia and Central were used as antiseptic and healing herbs. The Asia. In Turkey, the genus consists of 22 taxa, main components are natural flavonoids and 1361 Özek G, Özbek MU, Arslan M. JOTCSA. 2018; 5(3): 1361-1370. RESEARCH ARTICLE essential oils (Table 1), which are widely used the best of our knowledge, there is no previous as antiinflammatory, antibacterial, information about chemical composition and antispasmodic, and sedative agents (16). To biological activity of C. hamzaoglui. Table 1. Chemical composition of the essential oils of Cota species (literature survey) Plant Cota species Compound, (%) Ref. part C. altissima (L.) J. Gay (syn. -Pinene (4.0), benzaldehyde (27.1), -carene AP (17) Anthemis altissima L.) (4.2), linalool (4.6), β-caryophyllene (7.6) Decanoic acid (6.1), -caryophyllene (25.3), - Fl humulene (5.2), germacrene D (6.9), (18) spathulenol (5.4), caryophyllene oxide (6.5) C. altissima (L.) J. Gay (syn. Anthemis altissima L.) Carvacrol (3.5), -caryophyllene (17.2), L (18) spathulenol (17.4), caryophyllene oxide (9.6) trans--Farnesene (2.6), pentadecanoic acid St (18) (3.1), palmitic acid (39.6), linoleic acid (36.2) Cota palestina Kotschy (syn. Benzaldehyde (0.3-13.8), p-cymene (4.2-11.2), A. melanolepis Boiss.; chrysanthenol (3.3-4.4), benzyl alcohol (0- AP (17) Anthemis palestina (Reut. 26.9), 2-phenyl-1-ethanol 33.6), trans-verbenol Ex. Kotschy) Boiss.) (3.6-10.0), caryophyllene oxide (1.5-5.7) Cota triumfetti (L.) J. Gay -Eudesmol (18.2), borneol (13.3), (syn. Anthemis triumfetti (19) AP hexadecanoic acid (9.5), -eudesmol (8.6%), (L.) DC; A. talyshensis A. elemol (7.6) Fedor.) Cota tinctoria (L.) J. Gay 1,8-Cineole (7.9), -pinene (7.3), Fl (20) (syn. A. tinctoria L.) decanoic acid (5.4), -pinene (4.4) Cota triumfetti (L.) J. Gay -Pinene (16.9), camphor (15.0), -pinene AP (21) (syn. A. triumfetti (L.) DC.) (14.4), 1,8-cineole (5.8) AP: aerial parts; Fl: flowers; L: leaves; St: steams; syn: synonymous Recently, a new species Cota hamzaoglui Özbek & Vural in Anthemideae tribe has been Chemicals described in Turkey. Information given on the Boron trifluoride reagent (BF3), hydrochloric new species includes comments on the acid, n-hexane (Sigma-Aldrich, Germany), species’ affinity to Cota oxylepis Boiss. and C. calcium chloride, anhydrous sodium sulfate fulvida (Grierson) Holub (2). Several aspects (Fluka, Germany), diethyl ether (JT Baker, on chemical and pharmacological potent of the Holland), chloroform (Sigma-Aldrich, France), genus Anthemis have recently been reported methanol (Sigma-Aldrich, Poland) were of by Siasar-Karbasky et al. (22). A previous analytical grade. A C8–C40 n-alkane standard phytochemical studies on Anthemis species solution was purchased from Fluka (Buchs, resulted with polyphenols (23, 24), mono- and Switzerland). sesquiterpenes, fatty acids (25). Biological activity investigations of Anthemis species Instrumentation encompasses antibacterial (26), antioxidant Agilent 5975 GC-MSD system (Agilent, USA; (27), cytotoxic (28), antiproliferative (29), SEM Ltd., Istanbul, Turkey) was equipped with antidiabetic (30), antiinflammatory (31) and the HP-Innowax FSC column (60 m × 0.25 mm lipoxygenase inhibition (32) potentials. id with 0.25 m film thickness, Agilent, USA). The GC-FID analysis was carried out with In scope of the present work, we attempted to capillary GC using an Agilent 6890N GC investigate chemical composition of the system (SEM Ltd., Istanbul, Turkey). essential oil as well as fatty acid compositions of C. hamzaoglui. We have extracted the fatty Plant Material acids with Folch method (33) for subsequent The aerial parts of C. hamzaoglui were analysis of their composition after methylation collected on Bursa: Uludağ, above hotels, with boron trifluoride reagent (BF3). So, the between cable cars, and near an old tungsten present work is the first comprehensive mine, 2050-2100 m, 31.07.2009, U. Özbek investigation of the lipids and essential oil 2812 & M. Vural, and dried under the shade. constituents from aerial parts of C. Botanical identification was performed by Dr. hamzaoglui. M. U. Özbek. The voucher specimen is keep in the Herbarium of Gazi University, under MATERIALS AND METHODS herbarium code GAZI. 1362 Özek G, Özbek MU, Arslan M. JOTCSA. 2018; 5(3): 1361-1370. RESEARCH ARTICLE ratio of 40:1. The injector temperature was Hydrodistillation of Essential Oil 250 °C. Mass spectra were taken at 70 eV and The flowers and the herb of C. hamzaoglui the mass range was from m/z 35 to 450. were subjected to hydrodistillation (3 h) to yield essential oils in Clevenger-type Gas Chromatography – Flame Ionization apparatus according to European Detection (GC-FID) Pharmacopeia (34). The oil was dried over The GC-FID analysis was carried out with anhydrous sodium sulfate and stored in sealed capillary GC using an Agilent 6890N GC vial in refrigerator (4 °C), until GC-FID and system (SEM Ltd., Istanbul, Turkey). Flame GC/MS analyses. The oil was dissolved in n- ionization detector (FID) temperature was set hexane (10 %, v/v) to conduct at 300 °C in order to obtain the same elution chromatographic determination of its order with GC/MS. Simultaneous injection was composition. performed using the same column and appropriate operational conditions. Isolation of Fatty Acids and Derivatization Identification and Quantification of The ground plant material was subjected to Compounds maceration with chloroform: methanol (2:1) Identification of the volatile constituents was at room temperature for 24 h. The extract was based on the following: (i) comparison of filtered and the residue material was GC/MS Relative Retention Indices (RRI) of the macerated twice (for 30 min) more with new compounds on polar column determined portions of the solvent. All filtrates were relative to the retention times of a series of n- combined and half of the solvent was alkanes (C8-C40), with those of authentic evaporated under vacuum in a rotary compounds or literature data; (ii) computer evaporator. Then, half amount of chloroform matching with commercial mass spectral was added into the extract. The obtained libraries: MassFinder software 4.0, Adams extract was washed (three times) with CaCl2 Library, Wiley GC/MS Library (Wiley, New solution (0.4%) in a separatory funnel. At the York, NY, USA) and Nist Library, and end of the procedure, the chloroform extract comparison of the recorded spectra with was filtered through anhydrous sodium sulfate literature data. Confirmation was also to remove moisture, and then chloroform was achieved using the in-house Başer Library of removed under vacuum. The dried extract was Essential Oil Constituents database, obtained subjected to saponification. To do this, the from chromatographic runs of pure crude extract was boiled in KOH-H2O-MeOH compounds performed with the same (1:1:8) solution for 2 hours in a refluxing equipment and conditions (Table 2).