Obtain Essential Oil From The Leaves Of Chrysanthemum Indicum Var.aromaticum And Antifungal Activity Of The Essential Oil Against Fusarium Spp.

Chrysanthemum indicum var.aromaticum a new variety of Chrysanthemum genus,was first discovered in Shennongjia,Hubei Province,China.as a kind of spices and medicinal plant resources with high economic value[1].Taking C.indicum var.aromaticum as material base,C.indicum var.aromaticum leaves essential oil was obtained by an improved solvent-less microwave assisted hydrodistillation method,essential oil composition was determinated using GC-MS(gas chromatography-mass spectrometry).Chlorogenic acid and linarin in the resdues after hydrodistillation were extracted using microwave assisted method,and determinated through HPLC(high performance liquid chromatography),Antifungal activity of essential oil against Fusarium spp.also investigated.Its main contents are as follows:1.The essential oil of C.indicum var.aromaticum which being extracted by solvent-less microwave assisted solvent and the composition were determined by GC-MS.The experimental apparatus was made of a round-bottom flask,an extraction tube and a condensation tube were used.The influences of process parameters were investigated by single factor tests and response surface Box-Behnken design optimization.Under the optimum conditions(2 mL/g of liquid-solid ratio,40 min of microwave irradiation time,540 W of microwave irradiation power),the highest yield of essential oil was obtained.Compared with the traditional method,the experimental method improved the yield of essential oil in a short time.In addition,the GC-MS analysis of essential oil showed that the microwave-assisted extraction with less solvent had the highest compound content,accounting for 98.86%,while the essential oil extracted by conventional steam distillation had 98.34%compounds.Myrteneyl acetate(21.72%)and cis-sabinol(19.23%)were the main compounds in the essential oil of C.indicum var.aromaticum leaves,which contained large number of terpenes and other compounds.2.Chlorogenic acid and linarin in the resdues after hydrodistillation were extracted using microwave assisted method and determinated through HPLC,ensure that the flow rate is 0.5 mL/min,the sample volume is 10 ?L,the UV detection wavelength is 328 nm,the column temperature is 30? the mobile phase and the liquid phase are 0.8%phosphate methanol-water-acetonitrile gradient elution conditions.The optimal response surface conditions were determined by single factor experimental results.According to the results of response surface Box-Behnken method,the optimal extraction conditions were as follows:liquid and solid ratio was34 mL/g,microwave irradiation power was 540 W,microwave irradiation time was20 min,and the extraction yields of chlorogenic acid and linarin were the highest,among which the extraction yields of chlorogenic acid and linarin was 27.36±0.35 mg/g and 9.21±0.23 mg/g,respectively.3.The antifungal activity test of the extracted essential oils of Chrysanthemum indicum var.aromaticum leaves showed that the minimum inhibitory concentration and the minimum antifungal concentration of the essential oil were 1.5 ?L/mL and 4 ?L/mL against Fusarium trichothecioides,and the minimum inhibitory concentration and the minimum antifungal concentration against F.culmorum were 2 ?L/mL and 5?L/mL.According to the experimental results,it can be known that the essential oil of C.indicum var.aromaticum leaves has inhibitory effect on F.trichothecioides and F.culmorum,but the sensitivity of F.trichothecioides to essential oil is stronger than that of F.culmorum.