The essential oil compositions of Rosmarinus officinalis L. leaves growing in Mersin, Turkey European Journal of Chemistry 11 (4) (2020) 370-376 European Journal of Chemistry ISSN 2153-2249 (Print) / ISSN 2153-2257 (Online) – Copyright © 2020 The Authors – Atlanta Publishing House LLC – Printed in the USA. This work is published and licensed by Atlanta Publishing House LLC – CC BY NC – Some Rights Reserved. http://dx.doi.org/10.5155/eurjchem.11.4.370-376.2048 European Journal of Chemistry View Journal Online View Article Online The essential oil compositions of Rosmarinus officinalis L. leaves growing in Mersin, Turkey Gun Binzet 1,*, Riza Binzet 2 and Hakan Arslan 3 1 Department of Chemistry, Faculty of Education, Mersin University, 33343, Mersin, Turkey gunbinzet@mersin.edu.tr (G.B.) 2 Department of Biology, Faculty of Arts and Science, Mersin University, 33343, Mersin, Turkey rbinzet@mersin.edu.tr (R.B.) 3 Department of Chemistry, Faculty of Arts and Science, Mersin University, 33343, Mersin, hakan.arslan@mersin.edu.tr (H.A.) * Corresponding author at: Department of Chemistry, Faculty of Education, Mersin University, 33343, Mersin, Turkey. e-mail: gunbinzet@mersin.edu.tr (G. Binzet). 10.5155/eurjchem.11.4.370-376.2048 Received: 21 October 2020 Received in revised form: 09 November 2020 Accepted: 22 November 2020 Published online: 31 December 2020 Printed: 31 December 2020 This study reports on the chemical compositions of the essential oil of Rosmarinus officinalis L. (Rosemary) grown in Mersin, Turkey. The essential oil of rosemary was obtained by hydrodistillation method, and the yield of rosemary oil was found to be about 1.2 % (v:w). The hydrodistilled volatile oil was analyzed by gas chromatography and mass spectrometry techniques. Forty-five components were identified in the essential oil of R. officinalis, which represented 100% of the total essential oils. The oxygenated monoterpenes content possessed the highest value, 64.78% of the oil, among which eucalyptol (33.15%) and camphor (10.31%) were the most abundant components. In addition, the oil contained mainly monoterpene hydrocarbons, sesquiterpene hydrocarbons, oxygenated sesquiterpenes, and diterpenes. The least amount of diterpenes were found in the content of the oil. Isopimara-9 (11),15-diene (0.14%) and α-springene (0.06%) were two compounds determined as diterpene compounds. GC-MS Mersin 1,8-Cineole Essential oil Hydro-distilation Rosmarinus officinalis Cite this: Eur. J. Chem. 2020, 11(4), 370-376 Journal website: www.eurjchem.com 1. Introduction Essential oils obtained from plants are called aromatic or etheric oils, and these liquids are often rich in aroma. Essential oils are extracted from plants by dry distillation, water vapor distillation, or mechanical extraction methods. Consumers’ positive perception of being environmentally friendly and natural instead of synthetic additives has increased their interest in essential oils and their applications in recent years [1]. Commonly known as rosemary, Rosmarinus officinalis L., belonging to the Lamiaceae family, is a pleasant smelling woody aromatic perennial shrub that grows in Mediterranean areas and is widely distributed in many parts of the world [2-5]. Forms range from upright to trailing; the upright forms can reach 1.5 m, rarely 2 m. The leaves are 10-25 × 1-2(-4) mm, dark green, rugulose, and pilose above, white tomentose beneath. In addition, there are glandular hairs on the upper and lower surfaces of the leaf. The essential oil is stored in the secretion hairs on the epidermis [6]. It is known as biberiye, pürem, akpüren, hasalban, kuşdili, and urum çiçeği in Turkish [7,8]. In Turkey, R. officinalis is found growing on scrub areas, hillsides, and dry rocky slopes, in pine forests and particularly in the Mediterranean region, from just above sea level to 1.000 m [9]. Rosemary leaves are commonly used for flavoring foods, also this plant has also been widely used for different ethno- botanical and medicinal purposes. In traditional medicine, rosemary has been used as a stimulant and mild analgesic, and it has been considered as one of the most effective herbs for treating headaches, inflammatory diseases, poor circulation, and physical and mental fatigue. Rosemary has also been used empirically as a choleretic and hepatoprotective agent in folk medicine [5, 10,11]. Essential oil of R. officinalis is important for its medicinal uses and its powerful antibacterial, antioxidant, and antipro- liferative properties [12]. Likewise, the oil was characterized by its biological activities like antimicrobial [7,13], insecticidal [14,15], antioxidant [16], and anticancer properties [17]. R. officinalis essential oil is a colorless or pale-yellow liquid with the characteristic odor of the plant. Many studies on rosemary essential oil compositions have been reported. ABSTRACT RESEARCH ARTICLE KEYWORDS http://dx.doi.org/10.5155/eurjchem.11.4.370-376.2048 http://www.eurjchem.com/ http://dx.doi.org/10.5155/eurjchem.11.4.370-376.2048 mailto:gunbinzet@mersin.edu.tr mailto:rbinzet@mersin.edu.tr mailto:hakan.arslan@mersin.edu.tr mailto:gunbinzet@mersin.edu.tr http://www.eurjchem.com/ https://crossmark.crossref.org/dialog/?doi=10.5155/eurjchem.11.4.370-376.2048&domain=pdf&date_stamp=2020-12-31 Binzet et al. / European Journal of Chemistry 11 (4) (2020) 370-376 371 2020 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.11.4.370-376.2048 (a) (b) Figure 1. (a) Habitus and (b) map of the distribution of R. officinalis. The compositions of rosemary essential oil from different studies revealed the occurrence of the consists mostly of monoterpenes such as 1,8-cineole, α-pinene, camphor, borneol, camphene, α-terpineol, linalool, verbenane and bornyl acetate [5,18-34]. In previous studies, rosemary essential oil content and rates have varied. These variations were mostly associated with differences in the chemical composition of oils based on their distribution areas, environmental and agronomic conditions, collecting time, development stages of plants, and extraction method [22,35-38]. This study aims to determine the essential oils of R. officinalis wild in Mersin (Turkey) in order to compare the results with previously studies on the same plants. 2. Experimental 2.1. Chemicals All chemical solvents and reagents used were of analytical reagent grade and purchased from Merck, Darmstadt, Germany. 2.2. Plant materials The R. officinalis was collected by Dr. Riza Binzet from Mersin (Location: C5 Mersin, Mersin University Campus, damp slopes and scrub area, 14 May 2019, 36° 47ʹ 53ʺ N 34° 21ʹ 47ʺ E, 110 m, Binzet 201944 (Figure 1). The plant was identified by Dr. Riza Binzet. The voucher specimens are deposited in the Mersin University Research Herbarium (MERA), Mersin, Turkey. 2.3. Isolation of the essential oil Fresh leaves were harvested from wild plants of R. officinalis from the distribution area. The collected leaves were air-dried in the shade at room temperature (25 °C) for two months. Then, the dried leaves were powdered using a grinder (Blender 8011ES Model HGB2WTS3 400 W). In order to extract the essential oils, 50 g of the leaf powder was placed in a 1-liter round-bottomed flask with distilled deionized water (500 mL) and connected to the Clevenger apparatus. The steams in combination with the essential oils were distilled in to a graduated cylinder for 5 hours. The essential oil was separated from water, dried over anhydrous sodium sulphate, and stored prior to analysis in a sealed vial at +4 °C. They were analyzed in three weeks. 2.4. Analysis of essential oil 2.4.1. GC analysis The GC analysis was done using an Agilent Technologies 7890B GC. The carrier gas was helium at a flow of 3 mL/min. The column temperature was kept at 50 °C for 1 min and then heated to 250 °C with a 2 °C/min rate and kept constant at 280 °C for 10 min. The split ratio was 75:1 and the injector temperature was set at 280 °C. The purity of helium gas was 99.999 %. Essential oil samples (1 μL) were injected manually. 2.4.2. GC-MS analysis The GC-MS analysis was carried out with an Agilent 5977A GC-MSD system. The system was equipped with an automatic liquid sampler (Agilent 7693). HP-5MS 5% capillary column (coated with methyl silicone) (30 m × 250 μm × I.D., 0.25 μm) was used as the stationary phase. The temperature was programmed from 60 to 325 °C at a rate of 1°C/minute. The injector and interface temperatures were maintained at 325 and 350 °C, respectively. Mass spectra were taken at 70 eV. Mass range was from m/z 10 to 600. 2.5. Identification of components The qualitative identification of different compounds of the essential oil was performed on the basis of retention indices (RI) determined with reference to a homologous series of n- alkanes, under identical experimental conditions, co-injection with standards. Kovats index values were calculated according to the retention times and comparison of Kovats retention indices with literature values was carried out. The mass spectra were compared with those reported in the NIST14.L and W10N14.L computer libraries and those published in literature to date [39-53]. The minimum matching factor for decon- volution in The Automatic Mass Spectral Deconvolution and Identification System (AMDIS) is generally set above 90. 3. Results and discussion In this study, pale yellow coloured liquid essential oil of R. officinalis was obtained with the Clevenger apparatus with 1.20 % (v:w, volume:dry weight) yield. In many studies on rosemary, the essential oil has been reported by different researchers that it represents approximately 1-2.5% of the total weight and chemical composition of the plant [8,20,25,26,37,54-60]. The rate of essential oil obtained in this study and the rates of essential oil obtained in the above studies were found to be similar. The oil compounds identified by GC-MS analysis of R. officinalis qualitative and quantitative analyses results of essential oil were showed in Table 1 along with their Kovats indices and percentage composition. The GC-MS analysis of the essential oil of R. officinalis is shown in Figure 2. Representative mass spectra of essential oils extracted from R. officinalis are also given in Figure 3. The compounds are organized in order to their elution on HP-5MS column. The volatile compounds were identificated by comparing their mass spectra with Wiley library as well as with authentic compounds. 372 Binzet et al. / European Journal of Chemistry 11 (4) (2020) 370-376 2020 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.11.4.370-376.2048 Table 1. Chemical compositions of the essential oil extracted from R. officinalis as identified by GC-MS. No Compounds Rt 1 KIexp 2 KIlit 3 Percentage References 1 Tricyclene 6.959 914 919 0.20 [39] 2 3-Thujene 7.136 919 923 0.23 [40] 3 α-Pinene 7.409 927 933 8.11 [41] 4 Camphene 7.785 937 952 4.22 [40] 5 Sabinene 8.592 960 973 0.10 [41] 6 β-Pinene 8.727 964 964 3.60 [42] 7 β-Myrcene 9.315 981 991 1.48 [43] 8 S-2-Carene 9.729 993 993 0.07 [75] 9 o-Cymene 10.343 1010 1011 1.04 [44] 10 Eucalyptol (1,8-Cineole) 10.894 1026 1030 33.15 [45] 11 γ-Terpinene 11.906 1055 1059 0.69 [41] 12 α-Terpinolene 13.122 1090 1088 0.27 [43] 13 Linalool 13.548 1102 1098 0.75 [43] 14 Camphor 14.867 1140 1143 10.31 [43] 15 Borneol 16.161 1176 1165 7.16 [43] 16 Terpinen-4-ol 16.690 1192 1179 1.95 [40] 17 α-Terpineol 17.265 1208 1207 4.92 [40] 18 Geraniol 18.904 1255 1255 0.08 [43] 19 Bornyl acetate 21.576 1331 1302 5.66 [46] 20 Thymol 21.697 1334 1308 0.18 [46] 21 Carvacrol 21.923 1341 1314 0.13 [46] 22 Methyl eugenol 24.240 1321 1401 0.49 [43] 23 Isocaryophyllene 25.114 1432 1413 7.02 [41] 24 α-Caryophyllene 25.740 1450 1454 2.23 [43] 25 γ-Muurolene 26.182 1463 1477 0.25 [43] 26 β-Bisabolene 26.915 1483 1509 0.09 [43] 27 γ-Cadinene 26.962 1485 1512 0.24 [47] 28 trans-Calamenene 27.039 1487 1510 0.11 [48] 29 δ-Cadinene 27.182 1491 1524 0.41 [43] 30 Caryophyllene oxide 28.419 1526 1573 2.04 [41] 31 1,2-Epoxide-humulene 29.011 1543 1593 0.30 [48] 32 Methyl jasmonate 29.488 1557 1647 0.15 [43] 33 Alloaromadendrene 29.611 1560 1478 0.22 [49] 34 Caryophylla-4(12),8(13)-dien-5α-ol 29.708 1563 1602 0.32 [50] 35 Isoaromadendrene epoxide 30.290 1580 1579 0.80 [51] 36 Longiborneol 30.383 1583 1583 0.09 [76] 37 β-Caryophyllene 30.496 1586 1594 0.12 [52] 38 β-Caryophyllene oxide 30.631 1589 1581 0.20 [43] 39 α-Bisabolol 31.072 1602 1683 0.10 [43] 40 Cadalene 33.624 1676 1674 0.07 [43] 41 Germacrene B alcohol 34.319 1695 1694 0.09 [77] 42 α-Springene 37.392 1783 1781 0.06 [78] 43 Epi-Cryptomeridiol 37.719 1793 1790 0.08 [79] 44 Isopimara-9(11),15-diene 38.696 1820 1898 0.14 [53] 45 1S4R6R-g-Himachalen-4-yl acetate 39.216 1835 1830 0.08 [80] 1 Rt: Retention time. 2 KILit: Published Kovats retention indices. 3 KIexp: Kovats index determined experimentally relative to C8-C28 n-alkanes. In addition, the retention indices were calculated against n-alkane standards as reference. This was confirmed by comparison of their retention indices with those of authentic compounds as well as with data published in the literature [39- 53]. The percentage composition amounts were calculated from total ion chromatograms (TIC) by the computer. The GC-MS analyses of the obtained oil indicated the presence of forty-five volatile compounds, which included 100% of the total oil composition. The major compound of pale- yellow oil was found to be 1,8-cineole with 33.15 %. 1,8-Cineole is a natural organic monoterpenoid, also known as eucalyptol [61]. The name “Eucalyptol” is due to the fact that the major component of Eucalyptus oil is 1,8-cineole. The content of 1,8- cineole in eucalyptus oil varies from species to species, for example, it has been determined to be in high concentrations in Eucalyptus nicholii [62]. However, it is component of the essential oils of many plants. Eucalyptol is often used in food, fragrances, and cosmetics because of its fresh mint-like fragrance, spicy aroma, and taste [63]. Because of these properties, like many aromatic oils, 1,8-cineole is used in traditional medicine as a cough suppressor in bronchitis. Many researchers have studied the bioactive effect of rosemary essential oil, the main component of which is 1,8-cineole [61]. In our study, the other components in essential oil are camphor (10.31%), α-pinene (8.11%), isocaryophylene (7.02%), bornyl acetate (5.66%), α-terpineol (4.92%), camphene (4.22%), β- pinene (3.60%), α-caryophylene (2.23%), caryophyllene oxide (2.04%), terpinen-4-ol (1.95%), β-myrcene (1.48 %) and o- cymene (1.04%). In this study, the chemical profiles were marked by the presence of high amounts of oxygenated monoterpenes (64.78%) followed by monoterpene hydrocarbons (20.01%) (Table 2). The most abundant oxygenated monoterpene compounds identified in the oil were 1,8-cineole (33.15%) and camphor (10.31%). The sesquiterpene fraction (15.01%) was mainly composed of sesquiterpene hydrocarbons (10.54%), with isocaryophyllene (7.02%) being the main compound. Caryophyllene oxide (2.04%) was the most abundant of the six oxygenated sesquiterpenes identified. Isopimara-9 (11), 15- diene (0.14%), and α-springene (0.06%) were determined as diterpene compounds. In recent years, the chemical content of rosemary from different Mediterranean regions were examined by many researchers [8,17,25,27,35,36,37,57,60]. The chemical contents of essential oils of rosemary samples collected from different regions of Mersin province were investigated by a few researchers [7,8,36,63-65]. Essential oil components and their relative proportions were found to be different in this region. Generally, in Mersin, 1,8-cineole compound was determined as the main component. However, the 1,8-cineole percentage rates defined in essential oils were found to be different from each other [36,63-65]. Binzet et al. / European Journal of Chemistry 11 (4) (2020) 370-376 373 2020 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.11.4.370-376.2048 Figure 2. Profiles of GC-MS analysis for essential oil extracted from R. officinalis. Celiktas et al. [36], the percentage of 1,8-cineole was found highest with a rate of 50.7-61.4%. Similarly, in our study, 1,8- cineole was found as the highest main component with a value of 33.15%. On the other hand, Bagci et al. [8] in their study, camphor was identified as the highest main component with 14.48%, while Ozcan et al. [7] determined p-cymene as the main component with 44.02%. In our study, camphor was determi- ned as the second main component with 10.31%, while p- cymene was not found. In the analysis of essential oil compounds made with rosemary plants in Mersin, the 1,8- cineole, camphor, α-pinene and β-pinene compounds were defined as common compounds. According to the literature survey, the quantitative composition and the relative pro- potions of the oil’s components are widely influenced by environmental factors, soil characteristics, growing conditions and altitude [7,8,36,63-65]. In the study conducted by Celiktas et al. [36], the R. officinalis was collected from three different regions (Mersin, Canakkale, and Izmir) at four different time (December, March, June, and September) intervals of the year. In their study, they determined the variation in the essential oil composition of rosemary seasonally and regionally. 374 Binzet et al. / European Journal of Chemistry 11 (4) (2020) 370-376 2020 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.11.4.370-376.2048 Table 2. Main classes and subclasses of compounds of the essential oil extracted from R. officinalis. Classes of compounds Percentage Monoterpenes 84.79 Monoterpene hydrocarbons 20.01 Oxygenated monoterpenes 64.78 Sesquiterpenes 15.21 Sesquiterpene hydrocarbons 10.54 Oxygenated sesquiterpenes 4.47 Diterpenes 0.20 Total 100.00 (a) (b) Figure 3. Mass spectra of (a) 1,8-Cineol (RT: 10.894) and (b) Camphor (RT: 14.867) in essential oils extracted from R. officinalis. The seasons affected the main components amount, but it was not seasonal correlation was determined in essential oil compositions. It was determined that the components and their percentage values of essential oil of rosemary in our study differ from the components and their percentage values of essential oil of rosemary collected by Celiktas et al. [36] from Mersin in March-June, 2019. According to the results of both studies, no regular increase or decrease in essential oil compositions was determined seasonally. Similarly, in Yildirim [66] (2018), the seasonal variation of the essential oil compositions of the rosemary plant was examined in Kahramanmaraş and could not detect any correlation between the seasons. However, α- fenchene, γ-3-carene, dehydro-1,8-cineole, limonene, β- ocimene, 5-methyl-3-heptanone, 3-hexenol, 1-octan-3-ol, α- camppholene aldehyde, phenylacetaldehyde and humulene compounds were determined in the study by Celiktas et al. while these compounds were not detected in our study. This showed us that the chemical composition of rosemary oil varies according to the geographical region where it was collected. In addition, it has been determined that the main components of essential oil change in studies conducted in different regions [36,66]. Camphor was found as the main component in the analysis of essential oil components of rosemary plants, which spread in Izmir, Aydin, Antalya, Adana, and Hatay provinces (growing in Ankara ecologic conditions) by Gurbuz et al. [63]. According to this study, the amount of camphor in essential oil increases in cold climate conditions. In the study at Balıkesir [67], Fethiye [68], and Kahramanmaraş [66], in the essential oil compositions of rosemary were determinated limonene, sabinene, and ocimene, while they were not found in our study. Essential oil of R. officinalis from Tunisia [56,60,68], Pakistan [11], Lebanon [57], and Belgrade [69,70], shows a high content of 1,8-cineole, while the essential oil of R. officinalis from Iran [58,71], Spain [72], Italy [20,73], Morocco [59] shows low content of this molecule, and yields a high concentration of α- pinene instead; essential oil of R. officinalis from Brazil [54], in its turn, has a high concentration of camphor. These data indicated that the chemical composition variation due to the geographical area, edaphic features, and altitude where the plant is collected. The essential oil composition shows phytochemical variations according to the different parts of the plant used. Yosr et al. [74] noticed that the essential oil of R. officinalis obtained from leaves had 1,8-cineole (35.8%) as the major compound, while caryophyllene (16.7%) was the main compound in stem-extracted oil. However, in the essential oil extracted from flowers, the predominant component was caryophyllene oxide (11.9%). Similarly, in our study, essential oil was obtained from the leaves of rosemary and 1,8-cineole was determined as the main component. Our findings are consistent with the results of the study conducted by Yosr et al. [74]. Binzet et al. / European Journal of Chemistry 11 (4) (2020) 370-376 375 2020 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.11.4.370-376.2048 The major compound of the essential oil of rosemary obtained in this study was 1,8-cineole; this compound is widely used in the food and pharmaceutical industries. Thus, it is predicted that increasing the essential oil rate in rosemary can provide an important economic income source potential for our region. 4. Conclusion The present study is outlined to probe the chemical compositions of the essential oil of R. officinalis collected from Mersin province in Turkey. The hydrodistilled volatile oil was analyzed by GC-MS technique. Forty-five volatile compounds were determined according to NIST14.L and W10N14.L and literatures. The major compound in the essential oil was found to be 1,8-cineole with 33.15 %. The other components in volatile oil are camphor (10.31%), α-pinene (8.11%), isocaryo- phylene (7.02%), bornyl acetate (5.66%), α-terpineol (4.92%), terpinen-4-ol (1.95%), camphene (4.22%), β-pinene (3.60%), α-caryophylene (2.23 %), caryophyllene oxide (2.04 %), β- myrcene (1.48 %) and o-cymene (1.04%), respectively. The results of this study and other literature results showed that there is no seasonal correlation between essential oil compo- nents and their quantities. Disclosure statement Conflict of interest: The authors declare that they have no conflict of interest. Author contributions: All authors contributed equally to this work. Ethical approval: All ethical guidelines have been adhered. Sample availability: Samples of the compounds are available from the author. ORCID Gun Binzet http://orcid.org/0000-0002-9601-9528 Riza Binzet http://orcid.org/0000-0003-0336-8305 Hakan Arslan http://orcid.org/0000-0003-0046-9442 References [1]. Bosnalı, S.; Ozdestan, O. O. Pamukkale J. Eng. Sci. 2019, 25(7), 846-853. [2]. Cui, L.; Kim, M. O.; Seo, J. H.; Kim, I. S.; Kim, N. Y.; Lee, S. H.; Park, J.; Kim, J.; Lee, H. S. Food Chem. 2012, 132, 1775-1780. [3]. Albuquerque, U. P.; Medeiros, P. M.; Almeida, A. L. S.; Monteiro, J. M.; Freitas, E. M.; Melo, J. G.; Santos, J. P. J. Ethnopharmacol. 2007, 114(3), 325-354. [4]. Ventura-Martlneza, R.; Rivero-Osornoa, O.; Gomeza, C.; Gonzalez- Trujanob, M. E. J. Ethnopharmacol. 2011, 137, 1528-1532. [5]. Al-Sereiti, M. R.; Abu-Amerb, K. M.; Sen, P. Indian J. Exp. Biol. 1999, 37, 124-130. [6]. Davis, P. H.; Davis, P. Flora of Turkey and the East Aegean islands. Vol.7. Edinburgh University Press, Edinburgh, 1982. [7]. Ozcan, M. M. and Chalchat, J. C. Int. J. Food Sci. Nutr. 2008, 59, 691-698. [8]. Bagci, Y.; Kan, Y.; Dogu, S.; Celik, S. A. Indian J. Pharm. Educ. 2017, 51(3), S470-S478. [9]. Akgul, A. Spice science and technology, Publ. No. 15, Turkish Association Food Technologists, Ankara, Turkey, 1993. [10]. Yu, M. H.; Choi, J. H.; Chae, I. G.; Im, H. G.; Yang, S. A.; More, K.; Lee, I. S.; Lee, J. Food Chem. 2013, 136, 1047-1054. [11]. Raskovic, A.; Milanovic, I.; Pavlovic, N.; Cebovic, T.; Vukmirovic, S.; Mikov, M. BMC Complement Altern. Med. 2014, 14, 225. [12]. Hussain, A. I.; Anwar, F.; Chatha, S. A. S.; Jabbar, A.; Mahboob, S.; Nigam, P. S. Braz. J. Microbiol. 2010, 41(4), 1070-1078. [13]. Fu, Y.; Zu, Y.; Chen, L.; Shi, X.; Wang, Z.; Sun, S. T. Phytother Res. 2007, 21, 989-994. [14]. Amri, I.; Hamrouni, L.; Hanana, M.; Jamoussi, B.; Lebdi K. J. Agric. Res. 2014, 74, 273-279. [15]. Al-Younis, F.; Al-Naser, Z.; Al-Hakim, W. Int. J. Chemtech. Res. 2015, 8(3), 1382-1390. [16]. Hendel, N.; Larous, L.; Belbey, L. Int. Food Res. 2016, 23(4), 1725-1732. [17]. Gezici, S.; Sekeroglu, N.; Kijjoa, A. Indian J. Pharm. Educ. 2017, 51, S498-S503. [18]. Daferera, D. J.; Ziogas, B. N.; Polissiou, M. G. J. Agric. Food Chem. 2000, 48, 2576-2581. [19]. Jiang, Y.; Wu, N.; Fu, Y. J.; Wang, W.; Luo, M.; Zhao, C. J.; Zu, Y. G.; Liu, X. L. Environ. Toxicol. Pharmacol. 2011, 32, 63-68. [20]. Pintore, G.; Usai, M.; Bradesi, P.; Juliano, C.; Boatto, G.; Tomi, F.; Chessa, M.; Cerri, R.; Casanova, J. Flavour Fragr. J. 2002, 17, 15-19. [21]. Salido, S.; Altarejos, J.; Nogueras, M.; Sanchez, A.; Luque, P. J. Essent. Oil Res. 2003, 15, 10-14. [22]. Satyal, P.; Jones, T.; Lopez, E.; McFeeters, R.; Ali, N.; Mansi, I.; Al-kaf, A. G.; Setzer, W. N. Foods 2017, 6, 1-15. [23]. Boelens, M. H. Perfumer Flavorist 1985, 10, 21-37. [24]. Flamini, G.; Cioni, P. L.; Catalano, S.; Morelli, I. Riv. Ital. EPPOS Num. Speciale 1992, 3(8), 21-24. [25]. Chalchat J. C.; Garry R. P.; Michet, A.; Benjilali, B.; Chabart, J. L. J. Essent Oil Res. 1993, 5, 613-618. [26]. Arnold, N.; Valentine, G.; Bellomaria, B. J. Essent. Oil Res. 1997, 9, 167- 175. [27]. Elamrani, A.; Zrira, S.; Berrada, M.; Benjilali, B. A. J. Essent. Oil Res. 2000, 12, 487-495. [28]. Lahlou, M.; Berrada, R. Flavour Fragr. J. 2003, 18, 124-127. [29]. Pino, J. A.; Estarron, M.; Fuentes, V. J. Essent. Oil Res. 1998, 10(1), 111- 112. [30]. Lawrence, B. M. Perfum. Flavor. 1995, 20(1), 47-54. [31]. Lawrence, B. M. Perfum. Flavor. 1997, 22(5), 71-83. [32]. Zargari, A. Medicinal Plants, Tehran University Press, Tehran, 71-76, 1990. [33]. Rao, L. J.; Singh, M.; Raghavan, B.; Abraham, K. O. J. Food Qual. 1998, 21, 107-115. [34]. Moghtader, M.; Afzali, D. Am-Euras. J. Agric. Environ. Sci. 2009, 5(3), 393-397. [35]. Ben Jemia, M.; Tundis, R.; Pugliese, A.; Menichini, F.; Senatore, F.; Bruno, M.; Kchouk, M. E.; Loisso, M. R. Nat. Prod. Res. 2015, 29, 213- 222. [36]. Celiktas, O. Y.; Kocabas, E. H.; Bedir, E.; Sukan, F. V.; Ozek, T.; Baser, K. Food Chem. 2007, 100, 553-559. [37]. Hcini, K.; Sotomayor, J.; Jordan, M.; Bouzid, S. Asian J. Chem. 2013, 25, 2601-2603. [38]. Lopez, P.; Sanchez, C.; Batlle, R.; Nerin, C. J. Agric. Food Chem. 2005, 53, 6939-6946. [39]. Kowalski, R.; Wolski, T. Flavour Fragr. J. 2005, 20, 306-310. [40]. Hognadottir, A.; Rouseff, R. L. J. Chromatogr. A. 2003, 998, 201-211. [41]. Choi, H. S. J. Agric. Food Chem. 2003, 51, 2687-2692. [42]. Engel, E.; Baty, C.; LeCorre, D.; Souchon, I.; Martin, N. J. Agric. Food Chem. 2002, 50, 6459-6467. [43]. Adams, R. P. Identification of essential oil components by gas chromatography/mass spectroscopy. Allured Publishing Corporation Carol Stream, IL, 60188, USA, 1995. [44]. Caredda, A.; Marongiu, B.; Porcedda, S.; Soro, C. J. Agric. Food Chem. 2002, 50, 1492-1496. [45]. Jordan, M. J.; Margaria, C. A.; Shaw, P. E.; Goodner, K. L. J. Agric. Food Chem. 2002, 50, 5386-5390. [46]. Sotomayor, J. A.; Martinez, R. M.; Garcia, A. J.; Jordan, M. J. J. Agric. Food Chem. 2004, 52, 5418-5424. [47]. Tellez, M. R.; Canel, C.; Rimando, A. M.; Duke, S. O. Phytochem. 1999, 52(6), 1035-1040. [48]. Cavalli, J. F.; Tomi, F.; Bernardini, A. F.; Casanova, J. Flavour Fragr. J. 2003, 18, 532-538. [49]. Southwell, I. A.; Russell, M. F. Phytochem. 2002, 59, 391-393. [50]. Tzakou, O.; Constantinidis, T. Biochem. Syst. Ecol. 2005, 33(11), 1131- 1140. [51]. Congiu, R.; Falconieri, D.; Marongiu, B.; Piras, A.; Porcedda, S. Flavour Fragr. J. 2002, 17, 239-244. [52]. Chung, T. Y.; Eiserich, J. P.; Shibamoto, T. J. Agric. Food Chem. 1993, 41, 1693-1697. [53]. Demetzos, C.; Angelopoulou, D.; Perdetzoglou, D. Biochem. Syst. Ecol. 2002, 30(7), 651-665. [54]. Porte, A.; Godoy, R. L. D. O.; Lopes, D.; Koketsu, M.; Gonçalves, S. L.; Torquilho, H. S. J. Essent. Oil Res. 2000, 12(5), 577-580. [55]. Angioni, A.; Barra, A.; Cereti, E.; Barile, D.; Coisson, J. D.; Arlorio, M.; Dessi, S.; Coroneo, V. and Cabras, P. J. Agric. Food Chem. 2004, 52, 3530-3535. [56]. Akrout, A.; Hajlaoui, H.; Mighri, H.; Najjaa, H.; Jani, H. E.; Zaidi, S.; Neffati, M. J. Essent. Oil Bear. Pl. 2010, 13(4), 398-411. [57]. Apostolides, N. A.; El Beyrouthy, M.; Dhifi, W.; Najm, S.; Cazier, F.; Najem, W.; AbouKais, A. J. Essent. Oil Bear. Pl. 2013, 16(2), 274-282. [58]. Alipour, M.; Saharkhiz, M. J. Biocatal. Agric. Biotechnol. 2016, 7, 271- 278. [59]. Bouyahya, A.; Et-Touys, A.; Bakri, Y.; Talbaui, A.; Fellah, H.; Abrini, J.; Dakka, N. Microb. Pathog. 2017, 111, 41-49. http://orcid.org/0000-0002-9601-9528 http://orcid.org/0000-0003-0336-8305 http://orcid.org/0000-0003-0046-9442 376 Binzet et al. / European Journal of Chemistry 11 (4) (2020) 370-376 2020 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.11.4.370-376.2048 [60]. Abada, M. B.; Hamdi, S. H.; Masseoud, C.; Jroud, H.; Bousshih, E.; Jemaa, J. M. B. S. Afr. J. Bot. 2020, 128, 18-27. [61]. Juergens, U. R. Drug Res. 2014, 64, 638-646. [62]. Sadlon, A. E.; Lamson D. W. Altern Med. Rev. 2010, 15, 33-47. [63]. Gurbuz, B.; Bahtiyarca, R.; Uyanik, M.; Rezaeieh, A. P. K. Ind. Crops. Prod. 2016, 88, 1-5. [64]. Isikber, A. A.; Alma, M. H.; Kanat, M.; Karci, A. Phytoparasitica 2006, 34(2), 167-177. [65]. Polat, U.; Yesilbag, D.; Eren, M. J. Biol. Environ. Sci. 2011, 5(13), 23-30. [66]. Yildirim, E. D. Int. J. Agric. Sci. 2018, 4(1), 33-38. [67]. Orhan, I.; Aslan, S.; Kartal, M.; Sener, B.; Baser K. H. C. Food Chem. 2008, 108, 663-668. [68]. Perez-Alonso, M. J.; Velasco-Negueruela, A.; Emin D. M.; Harmandar, M.; Esteban, J. L. Essent. Oil Res. 1995, 7, 73-75. [69]. Kadri, A.; Zarai, Z.; Chobba, I. B.; Bekir, A.; Gharsallah, N.; Damak, M.; Gdoura, R. J. Med. Plants Res. 2011, 5(29), 6502-6508. [70]. Lakusic, D. V.; Ristic, M. S.; Slavkovska, V. N.; Sinzar-Sekulic, J. B.; Lakusic, B. S. Chem. Biodivers. 2012, 9(7), 1286-1302. [71]. Ladan Moghadam, A. R. J. Essent. Oil-Bear. 2015, 18(6), 1490-1494. [72]. Santoyo, S.; Cavero, S.; Jaime, L.; Ibanez, E.; Senorans, F. J.; Reglero, G. J. Food Prot. 2005, 68(4), 790-795. [73]. Melito, S.; Petretto, G. L.; Chahine, S.; Pintore, G.; Chessa, M. Nat. Prod. Commun. 2019, 14(7), 1934578X19864005, 2-7. [74]. Yosr, Z.; Hnia, C.; Rim, T.; Mohamed, B. Ind. Crops Prod. 2013, 43, 412- 419. [75]. Karioti, A.; Hadjipavlou-Litina, D.; Mensah, M. L. K.; Fleischer, T. C.; Skaltsa, H. J. Agric. Food Chem. 2004, 52(26), 8094-8098. [76]. Couladis, M.; Tsortanidou, V.; Francisco-Ortega, J.; Santos-Guerra, A.; Harvala, C. C. Flavour Fragr. J. 2001, 16(2), 103-106. [77]. De Kraker, J. W.; Schurink, M.; Franssen, M. C. R.; Konig, W. A.; de Groot, A.; Bouwmeester, H. J. Tetrahedron 2003, 59(3), 409-418. [78]. Shang, C.; Hu, Y.; Deng, C.; Hu, K. J. Chromatog. A 2002, 942(1-2), 283- 288. [79]. Lowe, R. F.; Russell, M. F.; Southwell, I. A.; Robinson, C. J.; Day, J. J. Essent. Oil Res. 2007, 19(4), 342-344. [80]. Omura, H.; Noguchi, T.; Nehira, T. Nat. Prod. Res. 2015, 30(4), 406-411. Copyright © 2020 by Authors. This work is published and licensed by Atlanta Publishing House LLC, Atlanta, GA, USA. The full terms of this license are available at http://www.eurjchem.com/index.php/eurjchem/pages/view/terms and incorporate the Creative Commons Attribution-Non Commercial (CC BY NC) (International, v4.0) License (http://creativecommons.org/licenses/by-nc/4.0). By accessing the work, you hereby accept the Terms. This is an open access article distributed under the terms and conditions of the CC BY NC License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited without any further permission from Atlanta Publishing House LLC (European Journal of Chemistry). No use, distribution or reproduction is permitted which does not comply with these terms. Permissions for commercial use of this work beyond the scope of the License (http://www.eurjchem.com/index.php/eurjchem/pages/view/terms) are administered by Atlanta Publishing House LLC (European Journal of Chemistry). http://www.eurjchem.com/index.php/eurjchem/pages/view/terms http://creativecommons.org/licenses/by-nc/4.0 http://www.eurjchem.com/index.php/eurjchem/pages/view/terms 1. Introduction 2. Experimental 2.1. Chemicals 2.2. Plant materials 2.3. Isolation of the essential oil 2.4. Analysis of essential oil 2.4.1. GC analysis 2.4.2. GC-MS analysis 2.5. Identification of components 3. Results and discussion 4. Conclusion Disclosure statement ORCID References PrintField10: PrintField11: PrintField12: PrintField13: PrintField14: PrintField15: PrintField16: PrintField20: PrintField21: PrintField22: PrintField23: PrintField24: PrintField25: PrintField26: