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       【摘要】 
       ObjectiveTo analyze chemical constituents of the essential oil from three Artemisia annua L. grown in Jinsha county, Guizhou province, China.Methods GCMS analytical method was adopted.Results29 compounds were identified, in which Monoterpenes ［monoterpene (52.45% ～75.32%) and Sesquiterpene (24.68% ～47.55%)］ were predominant , Camphor, Lborneol, Copaene, Caryophyllene, beta.Bisabolene, Germacrene D, Germacrene B, ()Neoclovene(II), Isoaromadendrene epoxide and Lanceol, cis. ConclusionThe chemical composition of the essential oil from three Artemisia annua L. is consistent, although the content of each compound is different, these informations can help to study the relationship of biosynthesis path of terpenes and Artemisinin in the future.
       【关键词】  Artemisia annua L Essentia1 oi1 GCMS
       Artemisia annua L. is an annual  native herb  of Asia and has been used for many centuries for the treatment of fever and malaria. Many secondary metabolites of terpene peroxides have been isolated from A. annua L., such as artemisia ketone, artemisinic alcohol, arteannuin B and myrcene hydroperoxide［1,2］. Some of them also can be found in essential oil ［3］. The most famous terpene peroxide is Artemisinin. It is chemically an amorphane-type sesquiterpene endoperoxide and becoming an important plant-derived compound in the treatment of the chloroquine-resistant and cerebral malarias ［4］. Thus, a need for adequate supplies of artemisinin is essential. Nowadays, scientists are interested in genetically engineering about the pathway of artemisinin biosynthesis to get high-yield Artemisia annua plants［5］. Many technologies have been applied in the qualitative analysis of terpenoids in Artemisia annua L. NMR and MS are more common and welcome ［2,3］. GCMS is a very useful method to analyze essential oil of A . annua L［6］. Guizhou of China is one of mainly region of A . L , but systematic studies about the herb, especially the essential oil from A . annua L., has not been previously mentioned in such literatures. This paper reports the analytical results of A . annua L. essential oil by using GC-MS for the first time.
       1  Material and methods
       1.1  Plant Materials Three A. annua L. were grown in Jinsha county, Guizhou province, China. They were planted seeds in March 2006 and collected in August 2006, at an altitude of 200-400 meters. All samples were identified by Chen Deyuan, a professor of Guiyang Traditional Chinese Medical College and the herbarium deposited at Analysis and Testing Center, Guizhou Normal University.
       1.2  Extraction of essential oilFresh leaves were dried in air and milled into crude powder. The essential oil was extracted from the powder by steam distillation method described in the Chinese Pharmacopoeia (2005) ［7］, Dry leaves of A . annua L. (50 g) were distilled for 6h using a essential oil extractor ,  with 1.21 % , 1.13 %  and 1.07% rate yielding of the sample 1, sample 2 and sample 3, respectively. The essential oil refrigerated at -20℃ during storage.
       1.3  Phytochemical analysis by combined Gas ChromatographyMass Spectrometry (GC-MS)The essential oil was submitted to quantitative analysis in a Shimadzu GCMSQP2010. GC conditions: carrier gas, helium at flow rate of 0.5 ml·min-1; sample size, 2 μl injected using the splitless injection technology; fused capillary silica column OV1701 (30 m × 0.33 mm × 0.25 μm). Temperatures: injector: 260℃, detector: 280℃, column: 50℃, 10℃·min-1, 260℃(20 min). The MS was taken at 70 eV. The MS scan parameters included a mass range of m/z 10500, a scan interval of 0.5 s, a scan speed of 1 000 amu ·s-1, and a detector voltage of 1.20 kV. The main constituents were identified by comparison with the masspectrums from Wiley and Nist 98 spectrum library［8］.
       2   Results
       2.1  The gas chromatography chromatogram of essential oil from sample 1 , sample 2 and sample 3（see Fig 1 , Fig 2 and Fig 3）.
       Fg 1  The gas chromatography chromatogram of sample 1 essential oil（略）
       Fg 2  The gas chromatography chromatogram of sample 2 essential oil（略）
       
       Fig 3  The gas chromatography chromatogram of sample 3 essential oil（略）
       2.2  The chromatographic profile of three Artemisia annua L. reveals that it contains 29 volatile compounds. Identified components were listed in Table 1. The monoterpene and Sesquiterpene compounds were predominant in the oil of three A . annua L. In the oil of sample 1 , Camphor (41.89%), Lborneol (10.39%), Camphene (3.06%), Lavandulol (0.83%), cisVerbenol (0.58%), L-borneol (0.61%), and Artemisia ketone (0.55%) as major monoterpenic, and  major sesquiterpenic compounds identified were Caryophyllene(8.46%), germacreneD (5.52%), Cubenol (2.17%) and Caryophyllene oxide(2.35%). In the oil of sample 2 , Camphor (34.46%),  Lborneol (11.26%), Camphene (2.32%), trans3(10) Caren2ol (1.24%), 4Terpineol(0.34%), and Lavandulol (1.24%) as major monoterpenic constituents, and the major sesquiterpenic compounds  identified were germacrene D (10.89%), Germacrene B (9.21%), Cubenol (0.87%), Chamigrene (2.51%), beta.-Bisabolene (0.78%), Caryophyllene oxide (3.23%), Longifolenaldehyde (2.67%) and Copaene (1.27%). In the oil of sample 3, Camphor, cis-Verbenol and L - borneol are major monoterpene compounds with 20.44%, 10.27%, and 9.24%, respectively. Otherwise, Camphene (0.73%), trans  3(10)  Caren2 ol(1.79%), Lavandulo (1.02%), and 4-Terpineol (0.85%) were identified, respectively, as monoterpenic constituents. The major sesquiterpenic compounds identified were Caryophyllene (10.14%), Germacrene B(7.32%) Germacrene D 5.57%), (-)-Spathulenol(5.24%), Cubenol (0.87%), Caryophyllene oxide(3.52%), Chamigrene (1.02%), Copaene (2.39%) and Lanceol, cis (2.35%).
       3  Discussion and conclusion
       
       Among these compounds of A . annua L essential oil, monoterpene (52.45% - 75.32%) and Sesquiterpene (24.68% - 47.55%) were predominant. The Camphor, Lborneol, Caryophyllene, germacrene D , Eucalyptol, Camphene, germacrene B, Cubenol and Lanceol, cis are major terpenic constituents, which proportion are over 60% of the total essential oil in A.annua L .
       
       As a whole, the chemical composition of the essential oil from three Artemisia annua L. are consistent, although content of each components are prominently different. These differences have something with their growth conditions.
       
       The chromatographic profile of A. annua L reveals that type of  compounds are monoterpene, monoterpenic oxides, sesquiterpene, sesquiterpenic oxides and sesquiterpenic peroxides.  It speculated that biosynthesis path of Artemisinin (sesquiterpenic hydroperoxide) relate to volatile terpene of A. annua L. These information can help to study the relationship of biosynthesis path of terpenes and Artemisinin in the future.
       Tab 1  Table 1 Main Constituents of the essential oils of three A.annua L （略）
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