Chemical Composition of Oil from Eremostachys Macrophylla

Chemical Composition of Oil from Eremostachys Macrophylla Chemical composition of the essential oil from aerial parts  of Eremostachys macrophylla Montbr. Auch.  from Northeast of Iran Akhlaghi H Abstract The essential oil obtained by hydrodisstillation of the aerial parts of Eremostachys macrophylla Montbr. Auch., grown wild in Iran, was analyzed by GC and GC/MS. The colorless oils were obtained by hydrodistillation, using a Clevenger-type apparatus for three hours, from aerial parts in 0.18% yield (w/w). Fourty-four compounds representing 91.6% of aerial parts oil of Eremostachys macrophylla were identified. The main components of the oil were hexadecanoic acid (27.5%), ethyl linoleate (8.5%), 6-methyl-ÃŽ ±-ionone (8.0%), isobutyl phthalate (5.8%), ÃŽ ±-cadinol (4.7%) and germacrene D (4.3%). The oil was rich in nonterpenoids (56.0%) and among them, oxygenated nonterpenes (53.2%) predominated over nonterpene hydrocarbons (2.8%). Key Word Index Eremostachys macrophylla Montbr. Auch., Lamiaceae, essential oil composition, hexadecanoic acid Introduction The genus Eremostachys of the family Lamiaceae (alt. Labiatae) contains 15 species of perennial in Iran, and five of them are endemic (1,2). During the past decade, seven investigations have been carried out on the chemical composition of the essential oils of the genus Eremostachys. These studies analyze the fresh aerial parts of Eremostachys laciniata Bunge from Jordan (3), flowers, stems, and roots of Eremostachys laevigata from Iran (4), flower, leaf and stem of Eremostachys macrophylla Montbr. Auch., and aerial part and stem of Eremostachys labiosa from Iran (5), aerial parts of Eremostachys adenantha and Eremostachys macrophylla from Iran (6), aerial parts of Eremostachys macrophylla from Central Iran (7), aerial parts of Eremostachys laevigata Bge. From Iran (8) and aerial parts of Eremostachys laciniata Bge. from Iran (9). Phytochemical investigation on a few species of Eremostachys revealed the presence of vicarin, a new isoflavone from Eremostachys vicaryi (10), eremosides A-C, New Iridoid Glucosides from Eremostachys loasifolia (11), loasifolin, a new flavonoid from Eremostachys loasifolia (12), a new acidic iridoid glucoside (13), furanolabdane diterpene glycosides from Eremostachys laciniata (14), new iridoid glucosides from Eremostachys moluccelloides Bunge (15) and Eremostachiin: a new furanolabdane diterpene glycoside from Eremostachys glabra (16). Our study dealt with the analysis of the essential oils of aerial parts of Eremostachys macrophylla Montbr. Auch grown wild in northeastern Iran. Experimental Plant material: The plant material was collected during the flowering stage in May 2012 from northern Sabzevar in Khorasan Province, Iran, at an altitude of 1580 meters. A voucher specimen has been deposited in the herbarium of Research Center of Natural Resources, Sabzevar, Iran. Essential oil isolation. Air-dried aerial parts of E. macrophylla (100 g) were subjected to hydrodistillation in a Clevenger-type apparatus for three hours to produce colorless oils. The yield of total volatiles was 0.18% (w/w). The oils were dried over anhydrous sodium sulfate and stored in sealed vials at 4  °C before analysis. GC analysis. GC analysis was performed using a Shimadzu GC-9A gas chromatograph, equipped with a HP-5MS fused silica column (30 mÃâ€"0.25 mm i.d., film thickness 0.25  µm). The oven temperature was held at 50  °C for five minutes and then programmed to 250  °C at a rate of 3  °C/min. The injector and detector (FID) temperatures were 290  °C . Helium was used as carrier gas with a linear velocity of 32 cm/s. GC/MS analysis. GC/MS analysis was carried out on a Hewlett-packard 6890 gas chromatograph fitted with a fused silica HP-5MS capillary column (30mÃâ€"0.25 mm ; film thickness 0.32  µm) . The oven temperature was programmed from 60  °C to 220  °C at 6  °C/min . Helium was used as carrier gas at a flow rate of 1 mL/min. The chromatograph was coupled to a Hewlett-Packard 5973 mass selective detector with an ionization voltage of 70 eV. Qualitative and quantitative analyses. Constituents of the volatile oils were identified by comparison of their retention indices relative to C9-C21 n-alkanes and of their mass spectral fragmentation pattern with those reported in the literature (17) and stored in a MS library (Wiley 275). The quantification of the components was performed on the basis of their GC peak area data from the HP-5MS column separation. Results and discussion Because of the variable results obtained in previous studies and as a part of on-going work on the chemical analysis of oils obtained from the wild plants of Iran, we decided to re-investigate the oils of this specific plant. Hydrodistilled volatile oils from the crushed dry aerial parts of Eremostachys macrophylla Montbr. Auch. (Lamiaceae) from Sabzevar (Iran) was studied by GC and GC/MS. The air-dried aerial parts of the plant yielded 0.18% (w/w) oil. The oil was clear and colorless. Fourty-four components were identified in the aerial parts oil that contained 91.6% of the compounds. Table 1 lists formulas, percentages, and retention indices of identified compounds in the oil. As evident from the table , the main components are hexadecanoic acid (27.5%), ethyl linoleate (8.5%), 6-methyl-ÃŽ ±-ionone (8.0%), isobutyl phthalate (5.8%), ÃŽ ±-cadinol (4.7%) and germacrene D (4.3%). In this study, GC and GC/MS analysis method revealed monoterpenoid hydrocarbon (MH), oxygenated monoterpenes (OM), sesquiterpenoid hydrocarbons (SH), oxygenated sesquiterpenes (OS), nonterpenoid hydrocarbons (NH), diterpene hydrocarbon (DH) and oxygenated diterpene (OD) in the oil from the aerial parts of Eremostachys macrophylla. One monoterpene hydrocarbon (0.1%), five oxygenated monoterpenes (8.8%), thirteen sesquiterpene hydrocarbons (13.4%), six oxygenated sesquiterpenes (10.4%), seventeen nonterpene hydrocarbons (56.0%), one diterpene hydrocarbon (2.5%) and one oxygenated diterpene (0.4%) were detected in this oil. The data lead to a rank order of constituent groups: NH>SH>OS>OM>DH>OD>MH for the aerial parts oil. The main components in this oil were hexadecanoic acid (27.5%), ethyl linoleate (8.5%), 6-methyl-ÃŽ ±-ionone (8.0%), isobutyl phthalate (5.8%), ÃŽ ±-cadinol (4.7%) and germacrene D (4.3%). The oil consisted mainly of nonterpenes and relatively small fractions of other terpenoids. Also, oxygenated nonoterpenes (53.2%) predominated over nonterpene hydrocarbons (2.8%). However, in a previous study on volatile oil from aerial parts of Eremostachys macrophylla, among the thirty-five identified compounds that have been compromised to 92.9% of the oil, spathulenol (23.4%), hexadecanoic acid (13.5%) and caryophyllene oxide (9.3%) were the major ones (6), while in other report on the aerial parts of this plant, among the sixteen identified compounds that have been compromised to 96.4% of the oil; germacrene-D (47.1%), germacrene-B (17.8%), ÃŽ ³-elemene (9.1%), myrcene (6.7%), ÃŽ ²-elemene (2.7%), and ÃŽ ²-phellandrene (2.6%) have been the major ones (7). Also, we reported analysis of the essential oils from flowers, leaves and stems of Eremostachys macrophylla (5). The specimen had been collected at different place, time and altitude from current study. The major compounds in the flower oil of E. macrophylla were 1,8-cineol (19.0 %) and germacrene D-4-ol (10.6 %), whereas the leaf oil contained ÃŽ ±-pinene (30.0 %), 1,10-di-epi cubenol (22.7 %), elemol ( 13.3 %) and bornyl acetate (11.0 %). The stem oil of the plant consisted mainly of 1,10-di-epi cubenol (34.4%) and elemol (24.0 %). Evident from the above data, there are significant differences in the results of the current study with previous studies (6,7) for the aerial parts of E. macrophylla . These discrepancies are not entirely unexpected since hydrodistillation relates to the interactions of the oil constituents with water vapor. Of course, there may also be differences related to environmental conditions such as climate, altitude, collection time, ground composition of the sampling area and different growth stages such as pre-flowering, fresh flowering and air-dried-flowering stages. Conclusion The chemical composition of the essential oil of aerial parts from Eremostachys macrophylla Montbr. Auch. (Lamiaceae) growing in Sabzevar was investigated. This study showed considerable amounts of hexadecanoic acid (27.5%), ethyl linoleate (8.5%), 6-methyl-ÃŽ ±-ionone (8.0%). These major constituents were different from previous studies on the same species 5-7. These results demonstrated that the chemical composition of the essential oil of the same species can change depending on a variety of conditions, including climate, time of collection, and the ground composition of the sampling area besides of growth stages of plant. Acknowledgment We are grateful to Dr. V. Mozaffarian (Research Institute of Forests and Rangelands, Tehran) for identifying the plant materials. Table 1. Constituents of the essential oils from aerial parts of Eremostachys macrophylla obtained by hydrodistillation a No. compound Formula Percentage RRIb Class 1 Limonene C10H16 0.1 1031 MHc 2 4-Terpineol C10H18O 0.1 1179 OMd 3 Fenchyl acetate C12H20O2 0.4 1223 OM 4 Anethole C10H12O 0.1 1285 OM 5 ÃŽ ±-Copaene C15H24 0.2 1364 SHe 6 ÃŽ ²- Bourbonene C15H24 0.8 1385 SH 7 ÃŽ ²-Cubebene C15H24 0.1 1390 SH 8 Tetradecane C14H30 0.1 1400 NHf 9 ÃŽ ²-Caryophyllene C15H24 0.3 1418 SH 10 ÃŽ ±-Guaiene C15H24 0.1 1439 SH 11 Aromadendrene C15H24 0.6 1442 SH 12 ÃŽ ±-Humulene C15H24 2.0 1452 SH 13 (E)-ÃŽ ²-Farnesene C15H24 0.1 1457 SH 14 Germacrene D C15H24 4.3 1480 SH 15 ÃŽ ²-Ionone C13H20O 0.2 1488 OM 16 Bicyclogermacrene C15H24 0.7 1500 SH 17 ÃŽ ³-Cadinene C15H24 0.6 1515 SH 18 6-Methyl-ÃŽ ±-ionone C14H22O 8.0 1518 OM 19 ÃŽ ´-Cadinene C15H24 3.5 1522 SH 20 Cadina-1,4-diene C15H24 0.1 1533 SH 21 Germacrene D-4-ol C15H26O 0.6 1574 OSg 22 Spathulenol C15H24O 1.5 1578 OS 23 Caryophyllene oxide C15H24O 0.5 1583 OS 24 Humulene epoxide II C15H24O 1.7 1608 OS 25 Ï„-Muurolol C15H26O 1.4 1643 OS 26 ÃŽ ±-Cadinol C15H26O 4.7 1656 OS 27 Tetradecanoic acid C14H28O2 1.8 1760 NH 28 Octadecane C18H38 0.4 1800 NH 29 6,10,14-Trimethyl-2-Pentadecanone, C18H36O 1.7 1848 NH 30 2-Hydroxy-Cyclopentadecanone C15H28O2 0.4 1853 NH 31 Pentadecanoic acid C15H30O2 0.3 1867 NH 32 Isobutyl phthalate C16H22O4 5.8 1877 NH 33 Cyclohexadecane C16H32 0.3 1883 NH 34 16-methyl-Oxacyclohexadecan-2-one, C16H30O2 0.3 1943 NH 35 Sandaracopimara-8(14),15-diene C20H32 2.5 1969 DHh 36 di-Butylphthalate C16H22O4 0.9 1973 NH 37 Hexadecanoic acid C16H32O2 27.5 1977 NH 38 Eicosane C20H42 2.0 2000 NH 39 Heptadecanoic acid C17H34O2 0.4 2065 NH 40 Methyl linoleate C19H34O2 0.6 2084 NH 41 Phytol C20H40O 0.4 2111 ODi 42 (Z,Z)-9,12-Octadecadienoic acid C18H32O2 2.7 2136 NH 43 Ethyl linoleate C20H36O2 8.5 2164 NH 44 Octadecanoic acid C18H36O2 2.3 2172 NH Total identified 91.6 a The compounds have been arranged according to their retention indices on an HP-5 MS capillary column b Kovatz retention indices given in the literature c Monoterpene hydrocarbons d Oxygenated monoterpene e Sesquiterpene hydrocarbons f Nonterpene hydrocarbons g Oxygenated sesquiterpene hDiterpene hygrocarbon iOxygenated diterpene References 1. 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