Studies On Extraction, Purification, Identification Of Chemical Constituents, Antioxidant And Antimicrobial Activities Of The Essential Oil From Fructus Evodia

Evodia Rutaecarpa,a traditional Chinese drug,which is the fruit of Evodia rutaecarpa(Juss.)Benth.,Evodia rutaecarpa(Juss.)Benth.var.officinalis(Dode)Huang and Evodia rutaecarpa(Juss.)Benth.var.bodinieri(Dode) Huang.The fruit of Evodia rutaecarpa,a Chinese traditional drug(Wu-Zhu-Yu),has been used in the treatment of headache,abdominal pain,migrains,chill limbs,postpartam hemorrhage,amenorrhage,dysentery,nausea and hyperbaropathy.Studies revealed that Evodia rutaecarpa have many pharmacological activities such as analgesic,sedative, antiseptic,anti-hypoxia and antihypertensive actions.Essential oil(volatile oil) is a kind of refining oil from plants,which has aroma and low boiling point.Volatile oil contains terpene and its oxides,their content are used to estimate the quality of raw materials.In order to research its biological active principles,further studies on the fruit has been undertaken.The extraction,chemical composition and bio-activities of essential oil from the fruit of Fructus Evodia were studied in this paper.1 Studies on extraction,purification of the essential oil from Fructus EvodiaThe essential oil from Fructus Evodia was purified,dissolved in the solvent,and analysised with gas chromatography(FID detector),the best analysis condition were: Column pressure 7.32psi,Heating rate 8℃/min.By comparing the spectrogram,the petroleum ether used as solvent was better than other extracting solvents.The RSD of relative retention＜1.92%;The RSD of chromatographic peak area＜2.36%.In addition,four different extraction methods, simultaneous distillation extraction,cold-soaked extraction,reflux extraction and ultrasonic extraction were compared,and the simultaneous distillation extraction was the best method. The extraction parameters including material/solvent,extraction time and crushing degree were optimized,the orthorgonal test results showed that the optimal extraction conditions were:material/solvent(g/mL) 1:14,extraction time 10h,crushing degree 20～40 mesh,under the optimal condition,the extracting rate of essential oil was 0.31%.2 Chemical composition of the essential oil from Fructus EvodiaThe chemical composition of the essential oil from Fructus Evodia was analyzed with GC-MS and the Kovats retention Indices(KI) methods.The results showed that more than 70 constituents were separated.Among which,29 constituents were identified by GC-MS and KI,accounting for 71%of the total amount of the oil.The main constituents in the essential oils from Fructus Evodia are 3-Carene,α-Pinene,Santolina triene,4-[1-Methylethyl]-1-cyclohexene-1-carboxaldehyde,Caryophyllene,Seychellene,γ-Selinene,(-)-Isoledene,1,4-Di-tert-pentylbenzene,Aristolene,δ-Cadinene,Santolina triene,Ledene oxide-(â…¡),d-Camphene,Pentadecanoic acid,methyl ester,α-Guaiene,(-)-Spathulenol,Cedren-13-ol,8-,Kaurene,1,2-Benzenedicarboxylic acid,butyl cyclohexyl ester,Isoearyophillene,Eugenol,Ocimene,6,9-Octadecadienoic acid,methyl ester and 1,3,6,10-Cyclotetradecatetraene,3,7,11-trimethyl-14-(1-methylethyl)-,[S-(E,Z,E,E)]-.3 Antioxidant activity of the essential oil from Fructus EvodiaThe K₃[Fe(CN)₆]system was used to evaluate the capacity of deoxidizing of Fructus Evodia volatile oil;by using the DNA system,superoxide radical(O₂^-·) system,hydroxyl (·OH) system and the peroxide(H₂O₂) system,the antioxidation activities of Fructus Evodia volatile oil were evaluated;the reagent kits were used to analyse the content of malondialdehyde(MDA) in mice liver homogenate and mice liver mitochondria;using spectrometer to evaluate swelling of mice liver mitochondria.The results showed that Fructus Evodia volatile oil was a good reducer and had evident inhibiting effects on O₂^-·,·OH and H_O2;in vitro,at concentration of 2.00mg/mL,the inhibiting rate of two extraction methods of Fructus Evodia volatile oil on radical-induced MDA formation were 65.38%and 61.54%respectively in mice liver mitochondria,21.43%and 31.90% (incubation),67.32%and 67.07%(incubation with Fe²⁺),49.54%and 47.43%(incubation with H₂O₂) respectively in mice liver homogenate;Fructus Evodia volatile oil made the radical-induced swelling of mice liver mitochondria much lower than that of mice liver mitochondria without inducement,there is no clear dose-effect relationship.In vivo, Fructus Evodia volatile oil significantly inhibited hepatic lipid peroxidation in liver tissue in a certain dose range.It could also promote the activity of superoxide dismutase(SOD) in blood serum.4 Antimicrobial activity of the essential oil from Fructus Evodia in vitroFilter paper measuration was adopted to estimate the antibacterial activity of Fructus Evodia essential oil.The results showed that T and H samples at concentration of 10mg/mL have certain degree of growth inhibition to Escherichia coli,Staphylococcus aureus and Pseudomonas aeruginosa,their antibacterial mechanism may be related to chemical components contained in the essential oil,such as Caryophyllene,Elemene, Eugenol,Pinene and Ocimene.