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Chemical Composition and Biological Properties of the Leaf Essential Oil of lucida Cav. from Cuba

Article in Journal of Essential Oil Research · September 2011 DOI: 10.1080/10412905.2011.9700485

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The user has requested enhancement of the downloaded file. Chemical composition and biological properties of the leaf essential oil of Tagetes lucida Cav. from Cuba

Erik L. Regalado and Miguel D. Fernández Center of Marine Bioproducts, Loma y 37, Havana, C.P. 10400, Cuba Jorge A. Pino* Food Industry Research Institute, Carretera al Guatao km 3½, Havana, C.P. 19200, Cuba. E-mail: [email protected] Judith Mendiola Institute of Tropical Medicine “Pedro Kourí”, P.O. Box 601, Havana, Cuba Olga A. Echemendia Institute Finlay. Ave 27 319805, Havana, C.P.11600, Cuba

Abstract The leaf essential oil of Tagetes lucida Cav. () from Cuba has been obtained by hydrodistillation and analyzed by GC-FID and GC–MS. Forty volatile compounds were identified, of which (96.8%) was the major constituent. The antioxidant capacity of this essential oil was measured by two different in vitro assays (DPPH and TBARS) and significant activities were evidenced. The preliminary screening of its antiplasmodial, antibacterial, antifungal and antiviral activities was carried out against Plasmodium berghei, , Pseudomonas aeruginosa, , , Acinetobacter lwoffi, Enterobacter aerogenes and against strains HHV 1 and HHV 2. The results showed a moderate activity against P. berghei and E. coli.

Key Word Index Tagetes lucida, Asteraceae, essential oil composition, estragole, antioxidant capacity, antiplasmodial activity, antimicrobial activity, antiviral activity.

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Introduction caused by helminthes (6). Moreover, a bibliographic Tagetes lucida Cav. (syn. T. florida Sweet, T. survey of plants for malaria in Latin America (2) reported schiedeana Less.), commonly called Pericón, hierbanís, the use of the dried powdered or the plant decoction anís, santa María, Mexican mint marigold, Mexican for the treatment of malaria in Mexico. , Spanish tarragon, or Texas tarragon, is a Essential oils from aromatic and medicinal plants have perennial herb that grows in dry rocky slopes and woods been known to possess important biological properties, native to Central America and South America and notably antibacterial, antifungal and antioxidant activities. naturalized elsewhere in the tropics and subtropics (1, 2). It Their biological potential depends on their chemical is cultivated commercially in Costa Rica as a spice herb; it composition determined by genotype and influenced by contains an essential oil having an -like odor, and the environmental and agronomic conditions (7). The chemical fresh aerial parts of this plant are sold in the supermarket composition of T. lucida volatile oil has been the subject of as a substitute of tarragon (3), which has been very used as previous studies. The major constituents of this volatile oil spice and to preserve meat (4). This species has been were determined to be methyl (80%) and estragole cultured in Cuba in gardens due to the beauty of its foliage, (12%) in México (8); estragole (45%) and methyl eugenol but it is not common its exploitation for medicinal aims. (20%) in Hungary (9); (23.8%), eugenol (24.3%) T. lucida has been referred in Mexican traditional and estragole (33.9%) in Guatemala (10) or estragole (95- medicine for different therapeutic applications. The 97%) in Costa Rica (3). T. lucida extracts have reported to infusion of leaves and flowers is drunk to combat diarrhea, act on bacteria and phytopathogenic fungi (5) and also rheumatism, asthma, and cold (5, 6). The decoction of the possesses antidepressant-like properties in rats (11). aerial parts is employed in the treatment of amoebic Moreover, considering both thiophene concentrations and dysentery, giardiasis, ascaridiasis and other infections biomass yields, T. lucida appeared to be a promising 2 Tagetes lucida Regalado et al.

species, with high potential for use as biocidal crops for substances or literature values (13), relative to C8-C32 n- the implementation of pest control practices (12). alkane series in a temperature-programmed run. In spite of the worldwide use of T. lucida in the folk 2,2-Diphenyl-2-picrylhydrazyl (DPPH) Radical medicine, the biological properties of its essential oil based Scavenging Assay: The antioxidant activity of the on experimental models have remained largely unexplored. essential oil was measured in terms of free-radical In this context, the present work describes a detail scavenging ability according to DPPH reported method chemical composition and examines the antiplasmodial, (14) with minor modifications. Basically, a 60-µM antibacterial, antifungal and antiviral activities of the methanolic solution of DPPH (980 µL) [Sigma-Aldrich essential oil isolated from the leaves of Tagetes lucida Co. (St. Louis, MO), prepared daily, was placed in a Cav. from Cuba. spectrophotometer cuvette, and eight concentrations of the essential oil of 0.1, 0.2, 0.4, 0.8, 1.2, 1.6, 2.0 and 3.0 Experimental mg/mL or ascorbic acid (standard) (0.16, 0.26, 0.6, 1.0 and Plant material: Leaves of T. lucida were collected in 1.30 mg/mL) in methanol (v/v) solution (20 µL) were February 2010, in the medicinal plants field of the Food added. The decrease in absorbance at 515 nm was Industry Research Institute in Havana, Cuba. The plant determined in a UV-1201 spectrophotometer, until the was identified by Dr. Pedro Herrera of the Institute of reaction plateau step was reached. Methanol was used to Ecology and Systematic (IES) and a voucher specimen zero the spectrophotometer. EC50 values were determined was deposited at the Herbarium of IES (HAC 44100). from the plotted graph of scavenging activity against Leaves (200 g) were submitted to hydrodistillation in a sample concentrations, which is defined as the total Clevenger-type apparatus for 2 hours. At the end of each antioxidant necessary to decrease the initial DPPH radical distillation the oils were collected, dried with anhydrous concentration by 50%. Triplicate measurements were Na2SO4, measured, and transferred to glass flasks that were carried out, and their scavenging effect was calculated filled to the top and kept at a temperature of −18°C for based on the percentage of DPPH scavenged. further analysis. Analyses were made by duplicate. Yields TBARS (thiobarbituric acid reactive species) assay: were calculated according to the weights of oils and plant The lipid peroxidation assay as TBARS was carried out material before distillation. according to a modified method (15). The reaction mixture Analysis of the essential oils: Oil sample analyses containing, in a final volume of 1.1 mL, 100 mL of were performed on A Konik 4000A instrument (Barcelona, cerebral tissue (whole brain) and 1 mL (0.05M) of Spain) equipped with a HP-5ms fused silica column (25 m KH2PO4/K2HPO4 buffer, pH 7.4 in NaCl (0.9%), and x 0.25 mm i.d., film thickness 0.25 µm), split injection seven concentrations of the essential oil (20, 50, 100, 150, 1:10, and flame ionization detection. Injector and detector 200, 250 and 500 µg/mL) was incubated at 37°C for 1 h. temperatures were at 220º and 250ºC. The oven Then, 1 ml of thiobarbituric acid (0.5%) and 1 mL of temperature was held at 70ºC for 2 min and then raised to acetic acid (20%) were added to the test tubes and were 250ºC at 4ºC/min and held for 10 min. The carrier gas was incubated at 100°C for 60 min. After cooling, absorbance H2 at 1 mL/min. Samples were injected by splitting and the was measured at 532 nm against control and buffer, BHT split ratio was 1:20. The lineal retention indices (RI) were being used as reference compound. All the experiments obtained from GC by logarithmic interpolation between were performed in triplicate, and the results were bracketing a homologous series of n-alkanes used as averaged. The inhibition percentage was determined by standards. Peak areas were measured by electronic comparison of the results between the samples and control. integration using the EZChrom Chromatography Data Antimalarial assay: In-vitro drug susceptibility was System 6.07 program (Scientific Software, Inc., FL). The determined in the standard short-term cultures of relative amount of the individual components was based Plasmodium berghei ANKA blood stages as described on the peak areas. before (16). Briefly, erythrocytes infected with parasites of GC/MS analysis was performed on a Shimadzu 17A P. berghei ring forms/young trophozoites are incubated at (Tokyo, Japan) gas chromatograph coupled to a Shimadzu 2% parasitemia at a final cell concentration of 1% in QP-5000 high performance quadrupole mass selective complete culture medium (RPMI 1640 with 20% Fetal detector was used. The GC was fitted with a HP-5ms fused Calf Serum, Sigma, St. Louis, MO) containing serial silica column (25 m x 0.25 mm i.d., film thickness 0.25 dilutions of essential oil maximal concentration tested, µm). The GC operating conditions were identical with from 200 µg/mL to 12.5 µg/mL, each in duplicate wells of those described above except that He was used as carrier 96-well culture plates. These plates are incubated for a gas. The MS operating conditions were: ionization period of 24 h at 37°C under standardized in vitro culture potential 70 eV with scan mass range of 35-400 m/z and conditions. The antimalarial activity was expressed as ion source temperature at 250ºC. Compounds were inhibitory concentration 50 (IC50), defined as the identified by computer search using digital libraries of concentration of the volatile oil that induces 50% reduction mass spectral data (NIST 02, Wiley 275, Adams 2001, of production of schizonts, which was calculated according Palisade 600, and Flavorlib home made library) and by to reported methodologies (17). IC50 was expressed as comparison of their retention indices of either reference mean ± standard deviation of tests performed in two Tagetes lucida Regalado et al. 3 different assays. Chloroquine diphosphate and artemisinin malonaldehyde (MDA), one of the secondary lipid (Sigma, St. Louis, MO) were used as references. peroxidation products, whose quantification gives a Antibacterial activity: The antibacterial activity was measure of the extent of lipid degradation. carried out by the diffusion method according to the For the first assay, solutions with eight volatile-oil National Committee for Clinical Laboratory Standard concentrations of 0.1–3 mg/mL, and different doses of Guideliness (NCCLS) (18) and evaluated against several ascorbic acid (positive control) were prepared to evaluate bacterial reference strains: Staphylococcus aureus (ATCC the DPPH radical-scavenging capacity. The respective 6538), Pseudomona aeruginosa (ATCC 9027), Escherichia scavenging capacities ranged from 10.2 ± 0.3% to 85.4 ± coli (10576), and Candida albicans (ATCC10231) and 1.1% with an EC50 value of 1.4 ± 0.3 mg/mL for the strains from clinical samples: Pseudomona aeruginosa, E. essential oil and 0.025 ± 0.004 mg/mL for ascorbic acid. coli, Acinetobacter lwoffi, and Enterobacter aerogenes. All For the second test, seven different concentration of the suspensions of microorganism were ajusted 0.5 volatile oil (20–500 µg/mL) and BHT as positive control, MacFarland and six concentrations of the essential oil also showed antioxidant activities in a dose dependent were tested. The plates were incubated to 37ºC in a manner and had 14.04 ± 0.09% to 96.26 ± 0.05% humidified atmosphere, containing 5% CO2 during 24 inhibition on lipid peroxidation. The IC50 value was found hours. to be 0.23 ± 0.03 mg/mL for the essential oil and 0.18 ± Cytotoxicity assay: Vero line was grown (37°C, 5% 0.04 µg/mL for BHT. CO2) in 96-well culture plates, in 199 medium, These results were in agreement with published data, supplemented with 10% fetal calf serum. Confluent which demonstrated that estragole is a weak DPPH radical monolayers were incubated 3 days with each dilution of scavenger (IC50 > 400 µM) (20). However, several the essential oil. All experiments were performed in investigations have demonstrated that some essential oils triplicate. The effect of samples on cell viability was (estragole chemotype) exhibit antioxidant potential [15, measured using the Naftol Blue Black (NBB) method (19). 16]. At the same time, the other significant component of Antiviral activity: The antiviral activity was evaluated this volatile oil, myrcene (2.3%) and a minor one (linalool, against strains HHV 1 and HHV 2 and performed in 96- 0.1 %) have been previously found to possess substantial well flat bottom tissue culture plates. Different dilutions of protective effect against oxidant induced genotoxicity, the essential oil were added to confluent monolayer of which is predominately mediated by their radical Vero cell. After 1 hour of incubation at 37°C in a 5% CO2, scavenging activity (21). In this context, the weak (DPPH) the virus suspensions were added. All plates were further and moderate (TBARS) antioxidant effects of T. lucida incubated at 37°C in a 5% CO2 atmosphere during 3 days. essential oil could be attributed in a great part to these volatile metabolites. Results and Discussion T. lucida essential oil exhibited a moderate antimalarial A yield of 0.79% (v/m) of leaf essential oil was activity, with IC50 value equal to 72 ± 3.62 µg/mL, which obtained by hydrodistillation of the fresh leaves of T. was evaluated following recommended endpoint criteria lucida. The chemical composition of the leaf oil is for natural complex mixtures (22. IC50 values for summarized in Table I. A total of 40 compounds were chloroquine (30.92 ng/mL) and artemisinin (18.3 ± 4.48 identified, accounting for 100% of the total composition. ng/mL) were significantly lower than this obtained for the Sixteen of them are reported for the first time. Estragole oil. In addition, they closely corresponded with previously (96.8%) dominated the leaf oil composition. The pleasant reported IC50 values of P. berghei ANKA (23). Previous sweet odor of T. lucida leaves is assumed to be caused by studies have underlined the potential biological activities the anise-like smelling estragole. Among all molecules of various essential oils against malaria parasites. At identified, the other quantified components included earliest research, eight essential oils were tested in vitro myrcene, germacrene D, (E)--ocimene, linalool, β- against P. falciparum and displayed IC50 values ranging caryophyllene and (E,E)--farnesene. from 149 to 1000 µg/mL (24), while more recently, other Previous studies on leaf oils classified T. lucida into five essential oils (Xilopia phloiodora, Pachypodanthium different chemotypes (3, 9, 10)] on the basis of the main confine, Antidesma laciniatum, Xylopia aethiopica, constituents. The leaf oil of T. lucida from Cuba could Hexalobus crispiflorus) (25) with IC50 values ranging from therefore be classified as the estragole high content type, 2 to 30 µg/mL were considered very active. Essential oils similar to the species reported in Costa Rica (3). Minor from fresh leaves of Cymbopogon citratus and Ocimum difference with the Costa Rican leaf oil is that other gratissimum growing in Cameroon showed significant like (E)-anethole and eugenol were antimalarial activities in the four-day suppressive in vivo found in trace in the Cuban leaf oil and they were not test in mice infected with Plasmodium berghei, at detected in Costa Rican leaf oil. concentrations of 200, 300 and 500 mg/kg of mouse To establish the antioxidant activity of this essential weight per day (26). In these reports, no clear conclusions oil, we used two well-established in vitro assays. The first can be derived from the chemical composition of the tested is based on the free-radical-scavenging capacity of the essential oils in association with their antimalarial activity. stable DPPH radical and the second concerns the The antioxidant activity of T. lucida essential oil is an spectrophotometric detection of TBARS, namely being interesting biological property. Endothelial cell injury by 4 Tagetes lucida Regalado et al. adherent parasitized red blood cells is ameliorated by using the diffusion method (18). This essential oil was not superoxide dismutase, ascorbic acid, or tocopherol in vitro, cytotoxic from 103 dilutions and did not show antiviral highlighting the potential therapeutic benefit of activity with strains HHV 1 and HHV 2. Estragol-rich antioxidants for severe malaria. At present, it is difficult to essential oils are reputed for their antifungal properties predict what will be their effect on malaria disease, but (30), but have also exhibited significant activity against future strategies might specifically target antioxidants to bacterial strains (31), including against E. coli (32). The the endothelium while keeping or enhancing oxidative antibacterial properties of T. lucida extracts have been stress in infected RBC (27). It will depend on the previously demonstrated, where some isolated coumarins distribution of the essential oil in the cells (28). were the most effective compounds against Gram-negative To the best of our knowledge, only extracts of flowers and Gram-positive bacterias (5). However, we report for of Tagetes erecta in ethanol/water and methanol/water the first time the activity of its essential oil against E. coli. solvent mixtures were evaluated in the in vitro antimalarial All bioactivities detected in T. lucida leaf essential oil are screening and against P. berghei in mice, which gave consistent with the worldwide ethnobotanical use of this negative results (29), so the moderate antimalarial activity medicinal plant. exhibited by T. lucida essential oil should be further explored. Acknowledgements Additionally, T. lucida essential oil only showed a The authors thank Dr Pedro Herrera for the moderate antibacterial activity against E. coli (10576) identification of the species.

References 1. M. P. Gupta, 270 Plantas Medicinales Iberoamericanas, Editora Presencia Ltda., Santafé de Bogotá, 157 (1995). 2. W. Milliken, Plants for malaria. Plants for fever. Medicinal species in Latin America- a bibliographic survey. The Royal Botanic Gardens, Kew; United Kingdom (1997). 3. J. F. Cicció, A source of almost pure methyl : volatile oil from the aerial parts of Tagetes lucida (Asteraceae) cultivated in Costa Rica. Rev. Biol. Trop. (Int. J. Trop. Biol.), 52, 853-857 (2004). 4. L. Kershaw, Edible & Medicinal Plants of the Rockies. Lone Pine, Edmonton, Canada (2000). 5. C. L. Céspedes, J. G. Avila, A. Martínez, B. Serrato, J. A. Calderón-Mugica and R. Salgado-Gaeciglia, Antifungal and antibacterial activities of Mexican tarragon (Tagetes lucida), J. Agric. Food Chem., 54, 3521-3527 (2006). 6. C. Márquez, O. F. Lara, R. B. Esquivel and E. R. Mata, Plantas Medicinales de México II. Composición, Usos y Actividad Biológica. Universidad Nacional Autónoma de México, México, 123 (1999). 7. F. Bakkali, S. Averbeck, D. Averbeck and M. Idaomar, Biological effects of essential oils – A review. Food and Chemical Toxicology, 46, 446–475 (2008). 8. A. Guzmán and A. Manjarrez, Estudio del aceite esencial de Tagetes florida. Bol. Inst. Quím. Univ. Nac. Autón. Méx., 14, 48-54 (1962). 9. E. Hethelyi, B. Danos, P. Tetenyi and G. Juhasz, Phytochemical studies on Tagetes species; infraspecific differences of the essential oil in T. minuta and T. tenuifolia. Herba Hungarica, 26, 145-158 (1987). 10. C. Bicchi, M. Fresia, P. Rubiolo, D. Monti, C. Franz and I. Goehler, Constituents of Tagetes lucida Cav. ssp. lucida Essential Oil. Flavour Fragr. J., 12, 47-52 (1997). 11. G. Guadarrama-Cruz, F. J. Alarcon-Aguilar, R. Lezama-Velasco, G. Vazquez-Palacios and H. Bonilla-Jaime, Antidepressant-like effects of Tagetes lucida Cav. in the forced swimming test. J. Ethnopharmacol., 120, 277-281 (2008). 12. I. Marotti, M. Marotti, R. Piccaglia, A. Nastri, S. Grandi and G. Dinelli, Thiophene occurrence in different Tagetes species: agricultural biomasses as sources of biocidal substances. J. Sci. Food Agric., 90, 1210-1217 (2009). 13. R.P. Adams, Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry, 4th ed. Allured Publ. Corp., Carol Stream, IL (2007). 14. W. Brand-Williams, M. E. Cuvelier and C. Berset, Use of a free radical method to evaluate antioxidant activity. Food Sci. Technol., 28, 25-30 (1995). 15. H. Ohkawa, N. Ohishi and K. Yagi, Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem., 95, 351-358 (1979). 16. C. J. Janse and A. P. Waters, Plasmodium berghei: the application of cultivation and purification techniques to molecular studies of malaria parasites. Parasitol. Today, 11, 138-143 (1995). 17. M. Schlichtherle, M. Wahlgren, H. Perlmann and A. Scherf, Methods in malaria research, Malaria Research and Reference Reagent Resource Center, Virginia (2000). 18. P.A. Wayne, Approved Standards MZ-8, Vol 25, NCCLS. 19. E. A. Gould and J. C. S. Clegg, Growth, titration and purification of alphaviruses and flaviviruses. Virology a practical approach, First Edition. Edits. B.W.J. Mahy pp. editorial? ciudad? (1985). 20. H. Tominaga, Y. Kobayashi, T. Goto, K. Kasemura and M. Nomura, DPPH radical-scavenging effect of several compounds and their glycoside derivatives. Yakugaku Zasshi 125 (4), 371-375 (2005). 21. D. Mitić-Ćulafić, B. Žegura, B. Nikolić, B. Vuković-Gačić, J. Knežević-Vukčević and M. Filipič, Protective effect of linalool, myrcene and eucalyptol against t-butyl hydroperoxide induced genotoxicity in bacteria and cultured human cells. Food Chem. Toxicol, 47, 260-266 (2009). 22. P. Cos, A. J. Vlietinck, D. Vanden Berghe and L. Maes, Anti-infective potential of natural products: How to develop a stronger in vitro “proof-of-concept”. J. Ethnopharmacol. 2006, 106, 290-302 (2006). Tagetes lucida Regalado et al. 5

23. B. Franke-Fayard, D. Djokovic, M. W. Dooren, J. Ramesar, A. P. Waters, M. O. Falade, M. Kranendonk, A. Martinelli, P. Cravo and C. J. Janse, Simple and sensitive antimalarial drug screening in vitro and in vivo using transgenic luciferase expressing Plasmodium berghei parasites. Int. J. Parasitol., 38, 1651-1662 (2008). 24. G. Milhau, A. Valentin, F. Benoit, M. Mallie, J. Bastide, Y. Pelissier and J. Bessiere, In vitro antimicrobial activity of eight essential oils. J. Essent. Oil Res., 9, 329-333 (1997). 25. F. F. Boyom, V. Ngouana, P. H. A. Zollob, C. Menut, J. M. Bessiere, J. Gut and P. J. Rosenthal, Composition and anti- plasmodial activities of essential oils from some Cameroonian medicinal plants. Phytochem., 64, 1269-1275 (2003). 26. F. Tchoumbougnang, P. H. Amvam Zollo, E. Dagne and Y. Mekonnen, In vivo antimalarial activity of essential oils from Cymbopogon citratus and Ocimum gratissimum on mice infected with Plasmodium berghei. Planta Med., 71, 20-23 (2005). 27. H. C. Ackerman, S. D. Beaudry and R. M. Fairhurst, Antioxidant therapy: Reducing malaria severity? Crit. Care Med., 37, 758-760 (2009). 28. F. Bakkali, S. Averbeck, D. Averbeck and M. Idaomar, Biological effects of essential oils – A review. Food Chem. Toxicol., 46, 446-475 (2008). 29. M. Misra, R. E. Rodriguez, S. L. North and K. S. Kasprzak, Nickel-induced renal lipid peroxidation in different strains of mice: concurrence with nickel effect on antioxidant defense systems. Toxicol. Lett., 58, 121-133 (1991). 30. S. Shin and C. A. Kang, Antifungal activity of the essential oil of Agastache rugosa Kuntze and its synergism with ketoconazole. Lett. Appl. Microbiol., 36, 111-115 (2003). 31. K. Carović-Stanko, S. Orlić, O. Politeo, F. Strikić, I. Kolak, M. Milos and Z. Satovic, Composition and antibacterial activities of essential oils of seven Ocimum taxa. Food Chem., 119, 196-201 (2010). 32. A. Guerrini, G. Sacchetti, M. Muzzoli, G. Moreno Rueda, A. Medici, E. Besco and R. Bruni, Composition of the volatile fraction of Ocotea bofo Kunth (Lauraceae) Calyces by GC-MS and NMR fingerprinting and its antimicrobial and antioxidant activity. J. Agric. Food Chem., 54, 7778-7788 (2006).

Table I. Chemical composition of Tagetes lucida leaf essential oil Compound RI % ethyl 2-methylbutanoate 851 tr (Z)-3-hexenol 859 tr myrcene 991 2.3 (Z)-3-hexenyl acetate 1005 tr (Z)--ocimene 1037 tr (E)--ocimene 1050 0.2 linalool 1097 0.1 methyl chavicol 1196 96.8 carvone* 1243 tr chavicol 1250 tr (E)-anethole 1285 tr -cubebene 1351 tr eugenol* 1359 tr -bourbonene* 1388 tr -elemene 1391 tr -caryophyllene 1419 0.1 -copaene* 1432 tr trans--bergamotene 1435 tr aromadendrene* 1441 tr (E)--farnesene 1457 tr germacrene D 1485 0.3 (Z,E)--farnesene* 1491 tr bicyclogermacrene 1500 tr -muurolene 1500 tr (E,E)--farnesene 1505 0.1 -cadinene 1523 tr elemol 1550 tr 1,10-di-epi-cubenol* 1619 tr epi--muurolol* 1642 tr -muurolol* 1646 tr -cadinol 1654 tr 14-hydroxy-9-epi-(E)-caryophyllene* 1670 tr n-nctadecane* 1800 tr hexahydrofarnesylacetone* 1844 tr n-nonadecane* 1900 tr n-eicosane* 2000 tr n-heneicosane 2100 tr phytol 2112 tr n-docosane* 2200 tr n-tricosane* 2300 tr * Reported for the first time in this essential oil. Trace constituent ( 0.1%).

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