Bangladesh Journal of Pharmacology Volume: 12; Number 4; Year 2017 Cite this article as: Sezen Karaoglan E, Özgen U, Kazaz C. Phytochemical studies on Origanum rotundifolium. Bangladesh J Pharmacol. 2017; 12: 470-471. Phytochemical studies on Origanum rotun- difolium Sir, The genus Origanum is one of the important genera of the Lamiaceae family. Origanum species have rich essential oil constituents and these plants are used as spices. They are also extensively used in the flavouring of food products and alcoholic drinks and perfumery for their spicy smell. Origanum species are used as antiseptic, stimulant, stomachic, expectorant, sudofiric and emmenagogic in the folk medicine (Aligiannis et al., 2001; Baser, 1978; Novak et al., 2000). Origanum rotundifolium is a member of Origanum genus. This medicinal plant known in Anatolia as "Mercan kösk isused as spices. Thymol, carvacrol, p-cymene and borneol were found to be as main constituents of the essential oil of O. rotundifolium. The essential oil of this plant shows various biological activities such as anti-oxidant, antibactial properties. These activities have been mainly attributed to constituents such as thymol, carvacrol, borneol, terpinenchemo type and spathulenol (Baser et al., 1993; Gormez et al., 2016; Goze et al., 2009; Kotan et al., 2010). In the present study, we report on the isolation and structure elucidation of 3 metabolites of O. rotundi- folium. The aerial parts of O. rotundifolium were collected from Barhal (Yusufeli, Artvin Province, 560-620 m, Turkey). A voucher specimen was deposited at the Herbarium of Ankara University, Faculty of Pharmacy (AEF25873). The air-dried and powdered aerial parts (400 g) of O. rotundifolium were extracted three times with MeOH at 40ºC (3 × 2L). After filtration, the methanol extracts were evaporated under vacuum to dryness. Methanol extract (71.6 g) was dissolved in water-methanol (9:1) and partitioned with chloroform and then ethyl acetate, which were separately concentrated and dried under reduced pressure to give 18.3 g and 4.0 g residues, respectively. The remaining aqueous phase was 31.0 g. Chloroform extract (2.7 g) was separated via silica gel column chromatography eluting with n-hexane-ethyl acetate (100:0, 90:10 ... 50:50). Fr. 6-7 gave the compound 1 (45 mg). Ethyl acetate extract was subjected to reversed phase silica gel column chromatography using water: methanol (90:10, 80:20…..,100) solvent systems. The fractions 4-10 (1.79 g) were further purified by consecutive column chromatography on silica gel (chloroform-methanol-water, 80:20:2, 70:30:3,…, 50:50:5) and sephadex LH-20 (MeOH), respectively. Fr. 8 gave compound 2 (44 mg). The remaining aqueous phase was subjected to revers- ed phase silica gel column chromatography using water: methanol (90:10, 80:20…..,100) solvent systems. Fr. 19-29 were separated via silica gel column chromatography eluting with chloroform-methanol- water (80:20:2, 70:30:3,…, 50:50:5). Fr. 84-121 gave compound 3 (217 mg). Their structures were identified by means of spectros- copic methods (1D- and 2D-NMR, EIMS). Compound 1 EIMS m/z236 [M+Na-H]+, C14H30O,1H-NMR (CDCl3, 400 MHz)δ: 3.66 (2H, t, J= 6.6 Hz, H-1), 1.28-1.60 (24H, H-2- 13), 0.90 (3H, t, J= 6.8 Hz, H-14). 13C-NMR (CDCl3, 100 MHz) δ:22.7, 25.7, 29.4, 29.4, 29.6, 29.7, 29.7, 31.9, 32.82 (C-2-13), 14.1 (C-14). Compound 2 C18H16O8,1H-NMR (CDCl3, 400 MHz)δ: 7.02 (1H, d, J= 1.8 Hz, H-2), 6.76 (1H, d, J= 8.4 Hz, H-5), 6.92 (1H, dd, J= 8.1, J= 2.2 Hz, H-6), 7.50 (1H, d, J= 16.0 Hz, H-7), 6.27 (1H, d, J= 16.0 Hz, H-8), 6.75 (1H, d, J= 1.8 Hz, H-2'), 6.67 (1H, d, J= 8.1 Hz, H-5'), 6.62 (1H, dd, J= 8.1 Hz, J= 1.8 Hz, H-6'), 3.09 (1H, dd, J= 14.1 Hz, J= 3.1 Hz, Ha-7'), 2.92 (1H, dd, J= 14.1 Hz, J= 9.7 Hz, Hb-7'), 5.08 (1H, dd, J= 9.7, J= 3.5 Hz, H-8'). 13C-NMR (CDCl3, 100 MHz) δ: 126.8 (C-1), 114.4 (C-2), 145.5 (C-3), 148.2 (C-4), 115.3 (C- 5), 121.7 (C-6), 145.5 (C-7), 113.9 (C-8), 167.9 (C-9), 129.9 (C-1 ), 116.3 (C-2 ), 144.8 (C-3 ), 143.6 (C-4 ), 115.0 (C-5 ), 120.6 (C-6 ), 37.6 (C-7 ), 76.3 (C-8 ), 176.1 (C-9 ). Compound 3 EIMS m/z 561 [M+Na]+ ,C27H22O12, 1H-NMR (CDCl3, 400 MHz)δ: 7.10 (1H, d, J= 8.4 Hz, H-6), 7.95 (1H, d, J= 16.0 Hz, H-7), 6.24 (1H, d, J= 16.0 Hz, H-8), 5.11 (1H, dd, J= 6.2 Hz, H-10), 3.07(bd, J= 13.0 Hz, Ha-11), 2.94 (dd, J= 13.0 Hz, J= 9.0 Hz, Hb-11), 6.87 (1H, s, H-13), 6.63 (d, J= 8.0 Hz, H-16), 6.57 (bd, J= 8.0 Hz, H-17), 4.26 (1H, d, J= 6.2 Hz, H-20), 5.87 (1H, d, J= 6.2 Hz, H-21), 6.85 (1H, s, H-23), 6.71-6.75 (3H, m, signaloverlap). 13C-NMR A Journal of the Bangladesh Pharmacological Society (BDPS) Bangladesh J Pharmacol 2017; 12: 470-471 Journal homepage: www.banglajol.info Abstracted/indexed in Academic Search Complete, Agroforestry Abstracts, Asia Journals Online, Bangladesh Journals Online, Biological Abstracts, BIOSIS Previews, CAB Abstracts, Current Abstracts, Directory of Open Access Journals, EMBASE/Excerpta Medica, Global Health, Google Scholar, HINARI (WHO), International Pharmaceutical Abstracts, Open J-gate, Science Citation Index Expanded, SCOPUS and Social Sciences Citation Index ISSN: 1991-0088; DOI: 10.3329/bjp.v12i4.34249 Letter to the Editor This work is licensed under a Creative Commons Attribution 4.0 International License. You are free to copy, distribute and perform the work. You must attribute the work in the manner specified by the author or licensor (CDCl3, 100 MHz) δ:123.6 (C-1), 129.0 (C-2), 147.5 (C-3), 145.3 (C-4), 116.6 (C-5), 119.9 (C-6), 142.7 (C-7), 116.0 (C- 8), 167.8 (C-9), 75.9 (C-10). At the end of the extraction and  isolation  processes  of  aerial  parts  of  O. rotundifolium  71.6 g  methanol  extract  was  obtained.  Chloroform,  ethyl acetate  and  remaining  aqueous phase were  18.3  g,  4 g  and  31 g,  respectively  after  fractionation. 1-Tetradecanol (Com- pound 1, 45 mg) was isolated  from  the chloroform  phase;  rosmarinic  acid (Compound 2, 44 mg) from  ethyl acetate  phase; lithospermic acid (Compound 3, 217 mg) fromthe aqueous phase (Figure 1). 1H-NMR and 13C-NMR data comply with the data given in the literature (Junges et al., 2000; Kelley et al., 1975; Kelley et al., 1976). Earlier studies have shown that O. rotundifolium has anti-oxidant, antibactial effects, including essential oils such as thymol, carvacrol, p-cymene and borneol (Gormez et al., 2016; Goze et al. 2009). The isolation of 1-tetradecanol, rosmarinic acid and lithospermic acid from O. rotundifolium was recorded for the first time in this study. Esen Sezen Karaoglan1, Ufuk Özgen2 and Cavit Kazaz3 1Atatürk University, Faculty of Pharmacy, Department of Pharmaceutical Botany, 25240 Erzurum, Turkey; 2Karadeniz Technical University, Faculty of Pharmacy, Department of Pharmacognosy, 61080 Trabzon, Turkey; 3Atatürk University, Faculty of Science, Department of Chemistry, 25240 Erzurum, Turkey. Corresponding author: email: esen.karaoglan@atauni.edu.tr References Aligiannis N, Kalpoutzakis E, Mitaku S, Chinou IB. Composition and antimicrobial activity of the essential oils of two Origanums pecies. J Agric Food Chem. 2001; 49: 4168- 70. Baser K, Ozek T, Tumen G, Sezik E.Composition of the essential oils of Turkish Origanum species with commercial importance. J Essent Oil Res. 1993; 5: 619-23. Baser KHC. The Turkish Origanum species. In: Oregano: The genera Origanum and Lippia. Kintzios SE (ed). London, Taylor and Francis, 1978, pp 109-26. Gormez A, Bozari S, Yanmis D, Gulluce M, Agar G, Sahin F. The use of essential oils of Origanum rotundifolium as antimicrobial agent against plant pathogenic bacteria. TEOP. 2016; 19: 656-63. Goze I, Alim A, Tepe AS, Sokmen M, Sevgi K, Tepe B. Screening of the anti-oxidant activity of essential oil and various extracts of Origanum rotundifolium Boiss. From Turkey. J Med Plants Res. 2009; 3: 246-54. Junges MJ, Fernandes JB, Vieira PC, Fernanades MDFGS, Filho ER, Frühauf M, Barañano AG. Triterpenosursânicos e oleanânicosisolados do caule de Eugeniaflorida DC. Revista de Pesquisa e Pós-Graduação. 2000; 1: 13-30. Kelley CJ, Harruff RC, Carmack M. 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Bangladesh J Pharmacol 2017; 12: 470-471 471 Figure 1: Compounds isolated from O. rotundifolium HO CH3 1 2 3 4 5 6 7 8 9 10 11 12 13 14 HO HO O OH O O OH OH 1 2 3 4 5 6 7 8 98' 9' 7' 1' 2' 3' 4' 5' 6' O O OH HO OH OH HO COOHO 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 2021 22 23 24 25 26 27 COOH 1 2 3 DatePrinted: This article was downloaded by you on: Dec 31, 2017