Study on the chemical constituents of the essential oil of the leaves ...

<strong>Study</strong> on the chemical constituents of the essential oil of the leaves of Eucalyptus globulus Labill / Asian Journal ofTraditional Medicines, 2009, 4 (4)The leaves of Eucalyptus globulus Labill werecollected from the Cangshan mountain region in theYunnan Province of China in May of 2007. Thespecimen was botanically identified by Professor JingmingJIA (School of Traditional Chinese MateriaMedica, Shenyang Pharmaceutical University,Shenyang). All solvents and reagents were ofanalytical grade.Extraction and isolation of essential oilThe fresh leaves were completely immersed inwater overnight, then water-distilled in a full glassClevenger-type apparatus to giving greenish-yellowoil. The extraction was carried out for 6 h andthe essential oil was dried over anhydrous sodiumsulphate and stored at 4 ºC before being analyzed.The extraction yield of this essential oil was 0.18 %(w/w).GC-MS analysis and identificationA Shimadzu GC-17A gas chromatographequipped with a GCMS-QP5050 column was used.This was coupled to a Mass spectrometer operated innegative chemical ionization mode. A fused-silicacapillarity column (30 m, 0.25 mm I.D., 0.25 mm filmthickness) with chemically bonded phases DB-1(J&WScientific) was used for the GC separation. The initialoven temperature was 60 ºC , this was held for 4 minthen raised at the rate of 4 ºC/min to 220 ºC, and thenheld for 15 min. Other operating conditions were asfollows: carrier gas, He (99.999 %), with a flow rateof 1 ml/min; injector temperature, 280 ºC; split ratio,10:1. Mass spectra were recorded at 70 eV and themass range was from m/z 33 to 500 amu.The constituents were identified by comparison oftheir mass spectra with those of NIST02 library datafor the GC-MS system (Fig. 2-5). The results werefurther confirmed by comparison of their retentionindices of the compounds with literature data [10, 11] .Results and discussionThe total ion chromatogram of the essential oil isdisplayed in Fig. 1. The amounts of the componentsfrom the volatile oil were determinated by thepeak area normalization method. This presence ofseveral overlapping peaks shows the complexityof the mixture. The chemical constituents of theessential oil are listed in Table 1. The essential oilconsisted mainly of oxygenated monoterpenes,monoterpenes and oxygenated sesquiterpenes. Ofthese, 1, 8-eucalyptol (72.71 %), α-terpineol (2.54 %),terpinen-4-ol (0.34 %), and linalool (0.24 %) werethe main oxygenated monoterpenes, while α-pinene(9.22 %), and β-pinene (0.4 %) were the mainmonoterpenes and α-eudesmol (0.39 %), (-)-globulol(2.77 %), and epiglobulol (0.44 %) were the mainsesquiterpene. Several significant compounds wereFig. 1. TIC of essential oil of Eucalyptus globulus Labill leavesFig.1 TIC of essential oil of Eucalyptus globulus Labill leaves135

<strong>Study</strong> on the chemical constituents of the essential oil of the leaves of Eucalyptus globulus Labill / Asian Journal ofTraditional Medicines, 2009, 4 (4)Table 1. Chemical constituents of the leaves essential oil of Eucalyptus globulus LabillNo. Retention time Molecular formula Compound Relative content (%)1 6.993 C 10H 16α-pinene 9.222 7.237 C 10H 16camphene 0.053 8.150 C 10H 16β-pinene 0.44 8.679 C 10H 16β-sabinene 0.255 9.113 C 10H 16limonene 0.046 10.785 C 10H 18O 1, 8-eucalyptol 72.717 11.012 C 10H 16cis-β-ocimene 0.038 11.324 C 10H 16γ-terpinene 0.119 11.595 - unidentified 0.0710 12.151 - unidentified 0.0711 12.362 C 10H 16terpinolene 0.1912 12.674 C 10H 18O linalool 0.2413 13.084 C 10H 18O fenchol 0.0514 13.282 C 10H 16O1, 7, 7-trimethylbicyclo [2.2.1] hept-5-en-2-ol0.0315 14.020 C 10H 16O pinocarveol 0.3616 14.563 C 10H 14O5, 5-dimethylene-3-hylenebicyclo [2.2.1]heptan-2-one0.1117 14.727 C 10H 18O 2, 6-dimethyl-1, 5, 7-octatrien-3-ol 0.0618 14.991 C 11H 18O 2isobornyl formate 0.1419 15.533 C 10H 18O terpien-4-ol 0.3420 16.156 C 10H 18O α-terpineol 2.5421 17.013 C 10H 16O trans-carveol 0.0922 17.258 C 10H 16O2-methylene-5-(1-methylethenyl)cyclohexanol0.0823 18.463 C 10H 18O 3, 7-dimethyl-2, 6-octadien-1-ol 0.1324 21.493 C 12H 20O 3exo-2-hydroxycineole 0.0425 22.073 C 12H 20O 2α-terpineol acetate 3.126 23.061 C 12H 20O 2geranyl acetate 0.7127 23.210 C 15H 24isoledene 0.0628 23.927 C 12H 20O 2isopulegol acetate 0.0429 24.443 C 15H 24α-gurjunene 0.130 24.685 C 12H 18O 2(-)-cis-carvyl acetate 0.0331 24.925 C 15H 24β-panasinsene 0.0332 25.158 C 15H 24β-gurjunene 0.0733 25.534 C 15H 24alloaromadendrene 2.4734 26.127 C 15H 24aromadendrene 0.6135 26.444 C 13H 18O 22-phenylethyl isovalerate 0.0936 26.850 C 15H 24eudesma-4(14), 11-diene 0.0537 26.999 C 15H 24α-guaiene 0.0738 27.235 C 15H 24(+)-ledene 0.2439 27.872 C 15H 22dehydroaromadendrene 0.0940 29.171 C 15H 26O epiglobulol 0.44136

<strong>Study</strong> on the chemical constituents of the essential oil of the leaves of Eucalyptus globulus Labill / Asian Journal ofTraditional Medicines, 2009, 4 (4)Continued table 141 29.311 C 15H 26O cubenol 0.0842 29.422 C 15H 26O ledol 0.0643 29.621 C 15H 24O spathulenol 0.1144 30.068 C 15H 26O (-)-globulol 2.7745 44 30.261 30.068 C 15 H 26 CO 15H 26O α-cadinol (-)-globulol 2.77 0.5746 30.490 C 15H 26O γ-eudesmol 0.2245 30.261 C 15 H 26 O -cadinol 0.5747 30.595 - unidentified 0.1248 44 31.084 30.068 15 26 C 15H 26O α-eudesmol (-)-globulol 2.7746 30.490 C 0.3915 H 26 O -eudesmol 0.2249 31.766 C 15H 26O β-eudesmol 0.0445 30.261 C 15 H 26 O -cadinol 0.5750 47 31.886 30.595 - C 15H 26O 1, unidentified 2, 3, 3a, 4, 5, 6, 7-octahyfro-α, 3, 8-t-5- 0.12 0.07azulenemethanol46 30.490 15 26 -eudesmol 0.2248 31.084 C 15 H 26 O -eudesmol 0.3947 30.595 - unidentified 0.12α-terpineol 49 acetate 31.766 (3.1 %), geranyl C 15 H 26 Oacetate (0.71 %), -eudesmol constituents are similar to those given 0.04 in the literature[10, 11]L-pinocarveol 48 (0.36 31.084 %), β-sabinene C 15 H 26 O (0.25 %), and -eudesmol . Some constituents, such as β-sabinene, 0.39 linalool,1,2,3,3a,4,5,6,7-octahyfro-,3,8-t-5-azulenemetterpinolene 50 (0.1931.886 %). A portion C (0.26 %) of the total isoledene, ledene, epiglobulol, and cubenol had no49 31.766 15 Hconstituents remains unidentified. 15 26 O0.0726 -eudesmol 0.04literature hanol data [10, 11] . This possibly may be due to theThe yield of essential oil and the content 1,2,3,3a,4,5,6,7-octahyfro-,3,8-t-5-azulenemetof fact that the trees grown in different regions may exhibit8-eucalyptol 50 are 31.886 within the C 15 values H 26 O reported in the differences in their chemical constituents. 0.07literature [12] and the percentage contents of the mainhanolAFig.2 The MS of - pinene in the essential oil of Eucalyptus globulus Labill leavesBFig.2 The MS of - pinene in the essential oil of Eucalyptus globulus Labill leavesFig. 2. A. The Fig.3 MS of The α-pinene MS of in - the essential pinene in oil NIST02 of Eucalyptus library globulus data of Labill the GC-MS leave; B. system The MS of α-pinene in NIST02 librarydata of the GC-MS systemFig.3 The MS of - pinene in NIST02 library data of the GC-MS system137

<strong>Study</strong> on the chemical constituents of the essential oil of the leaves of Eucalyptus globulus Labill / Asian Journal ofTraditional Medicines, 2009, 4 (4)AFig.4 The MS of 1,8-eucalyptol in the essential oil of Eucalyptus globulus Labill leavesFig.4 The MS of 1,8-eucalyptol in the essential oil of Eucalyptus globulus Labill leavesFig.4 The MS of 1,8-eucalyptol in the essential oil of Eucalyptus globulus Labill leavesBFig.4 The MS of 1,8-eucalyptol in the essential oil of Eucalyptus globulus Labill leavesFig.5 The MS of the 1,8-eucalyptol in NIST02 library data of the GC-MS systemFig. 3. A. The MS of 1, 8-eucalyptol in the essential oil of Eucalyptus globulus Labill leaves; B. The MS of the 1, 8-eucalyptol inFig.5 The MS of the 1,8-eucalyptol in NIST02 library data of the GC-MS systemNIST02 library data of the GC-MS systemFig.5 The MS of the 1,8-eucalyptol in NIST02 library data of the GC-MS systemAFig.5 The MS of the 1,8-eucalyptol in NIST02 library data of the GC-MS systemFig.6 The MS of alloaromadendrene in the essential oil of Eucalyptus globulus Labill leavesFig.6 The MS of alloaromadendrene in the essential oil of Eucalyptus globulus Labill leavesBFig.6 The MS of alloaromadendrene in the essential oil of Eucalyptus globulus Labill leavesFig.6 The MS of alloaromadendrene in the essential oil of Eucalyptus globulus Labill leavesFig.7 The MS of the alloaromadendrene in NIST02 library data of the GC-MS systemFig. 4. A. The MS of alloaromadendrene in the essential oil of Eucalyptus globulus Labill leaves; B. The MS of thealloaromadendrene in NIST02 library data of the GC-MS systemFig.7 The MS of the alloaromadendrene in NIST02 library data of the GC-MS systemFig.7 The MS of the alloaromadendrene in NIST02 library data of the GC-MS system138Fig.7 The MS of the alloaromadendrene in NIST02 library data of the GC-MS system

<strong>Study</strong> on the chemical constituents of the essential oil of the leaves of Eucalyptus globulus Labill / Asian Journal ofTraditional Medicines, 2009, 4 (4)AFig.8 The MS of (-)-globulol in the essential oil of Eucalyptus globulus Labill leavesBFig.8 The MS of (-)-globulol in the essential oil of Eucalyptus globulus Labill leavesFig. 5. A. The MS Fig.9 of (-)-globulol The MS of in (-)-globulol the essential in oil NIST02 of Eucalyptus library globulus data of Labill the GC-MS leaves; B. system The MS of (-)-globulol in NIST02library data of the GC-MS systemResults and DiscussionConclusion Fig.9 The MS of (-)-globulol in NIST02 library [3] data Gray of AM, the Abdel-Wahab GC-MS system YHA, Flatt PR. The traditionalplant treatment, sambucus nigra (elder), exhibits insulinlikeand insulin-releasingIn the The present total ion work, chromatogram 47 constituents of the of essential the oil is displayed in Fig.1. Theactionsamountsin vitro.of theJ Nutr, 2000,essential oil from Eucalyptus globulus Labill grown in 130(1): 15-20.components from the volatile oil were determinated by the peak area normalization method. Thisthe Cangshan Results and mountain Discussion region in the Yunnan Province [4] Ikawati Z, Wahyuono S, Maeyama K. Screening ofseveral Indonesian medicinal plants for their inhibitoryof China presence were successfully of several overlapping identified and peaks determined. shows the complexity of the mixture. The chemicalThe total ion chromatogram of the essential oil is displayed effects on histamine in Fig.1. The release amounts from RBL-2H3 of the cells. JA comparison with literature data showed that the Ethnopharmacol, 2001, 75(2-3): 249-56.constituents of the essential oil are listed in Table 1. The essential oil consisted mainly ofmajor components constituents from in the essential volatile oil oil were determinated similar by [5] the Cimanga peak area K, Kambu normalization K, Tona L, method. Apers S, This De Bruyne T,to those oxygenated reported monoterpenes, elsewhere, although monoterpenes there and were oxygenated sesquiterpenes.Hermans N, TottéOfJ,these,Pieters1,8-eucalyptolL, Vlietinck AJ. Correlationpresence of several overlapping peaks shows the complexity between chemical of the mixture. composition The and chemical antibacterial activitydifferences in the percentage composition. Some trace(72.71%), α-terpineol (2.54%), terpinen-4-ol (0.34%), and of essential linalool oils of (0.24%) some aromatic were medicinal the main plants growingor minor constituents chemical of constituents the essential have oil not are reported listed in in Table 1. in The the essential democratic oil republic consisted of Congo. mainly J Ethnophamacol, ofthe literature. oxygenated monoterpenes, while α-pinene (9.22%), and 2002, β-pinene 79(2): 213-20. (0.4%) were the mainoxygenated monoterpenes, monoterpenes and oxygenated [6] Osawa sesquiterpenes. K, Yasuda Of H. Macrocarpals these, 1,8-eucalyptol H, I and J from themonoterpenes and α-eudesmol (0.39%), (-)-globulol (2.77%), leaves and of Eucalyptus epiglobulol(0.44%) globulus. J Nat were Prod, the 1996, 59(9):References (72.71%), α-terpineol (2.54%), terpinen-4-ol (0.34%), 823-7. and linalool (0.24%) were the main[1] Liu main YM, sesquiterpene. Li SF, Wu YT. Several Advances significant in the study compounds of [7] were Dessi α-terpineol MA, Deiana acetate M, Rosa (3.1%), A, Piredda geranyl M, CottigliaEucalyptus oxygenated globulus monoterpenes, Labill. J Chin while Med Materials, α-pinene 2003, (9.22%), and F, Bonsignore β-pinene L, (0.4%) Deidda were D, Pompei the R, main Corongiu FP.26(6): acetate 461-3. (0.71%), L-pinocarveol(0.36%), β-sabinene (0.25%), Antioxidant and terpinolene activity of (0.19%). extracts A from portion plants growing in[2] Swanston-Flatt monoterpenes SK, Day and C, α-eudesmol Bailey CJ, Flatt (0.39%), PR. Traditional (-)-globulol (2.77%), Sardinia. and Phytother epiglobulol(0.44%) Res, 2001, 15(6): were 511-8. theplant (0.26%) treatments of the total for diabetes. constituents Studies remains normal unidentified. and [8] Yun BS, Lee IK, Kim JP, Chung SH, Shim GS, Yoo ID.streptozotocin main sesquiterpene. diabetic mice. Several Diabetologia, significant 1990, compounds 33(8): were Lipid α-terpineol peroxidation acetate inhibitory (3.1%), activity of geranyl some constituents462-4. The yield of essential oil and the content of 1,8-eucalyptol isolated are from within the stem the bark values of Eucalyptus reported in globulus. Archacetate (0.71%), L-pinocarveol(0.36%), β-sabinene (0.25%), and terpinolene (0.19%). A portionthe literature [12] and the percentage contents of the main constituents are similar to those given in(0.26%) of the total constituents remains unidentified.The yield of essential oil and the content of 139 1,8-eucalyptol are within the values reported in

<strong>Study</strong> on the chemical constituents of the essential oil of the leaves of Eucalyptus globulus Labill / Asian Journal ofTraditional Medicines, 2009, 4 (4)Pharm Res, 2000, 23(2): 147-50.[9] Luo JL, Song YF. Chemical constituents of the essential oilfrom the leaves of three species of Eucalyptus. Nat ProdRes Dev, 1991, 3(3), 79-83.[10] Li H, Madden JL. Analysis of Leaves Oils from aEucalyptus Species Trial. Biochem Syst Ecol, 1995,23(2): 167-77.[11] Silvestre AJD, Cavaleiro JAS, Delmond B, Filliatre C,Bourgeois G. Analysis of the variation of the essential oilcomposition of Eucalyptus globulus Labill from Portugalusing multivariate statistical analysis. Ind Crop Prod,1997, 6(1): 27-33.[12] Lassak EB. The Australian Eucalyptus oil industry, pastand present. Chem Australia, 1998, 55(11): 396-8.140