| Asarum heteropoides Fr.Schmidt var.mandshuricum(Maxim.)Kitag.is a valuable Chinese herbal medicine.Moreover,the roots and rhizomes of A.heterotropoides var.mandshuricum,it is widely distributed in Northeast China and partly distributed in Japan and Korea.In recent years,A.heterotropoides var.mandshuricum has been widely used in the pharmaceutical industry.After A.heterotropoides var.mandshuricum was officially included in the Chinese Pharmacopoeia,its application and research have attracted more and more attention from researchers.Based on the material basis of A.heterotropoides var.mandshuricum roots and rhizomes,this paper focuses on the important needs of national forestry economic and social development,and takes the content of asarinin and sesaminin the roots and rhizomes of A.heterotropoides var.mandshuricum as the research object,using microwave-assisted steam distillation followed solvent extraction method to obtain optimization of the extraction process by response surface design,the antibacterial experiment of the Fusarium spp.Its main contents are as follows:1.Using microwave-assisted steam distillation method to extract essential oil from the roots and rhizome of A.heterotropoides var.mandshuricum,experiments have shown that,under the condition of the moisture content of A.heterotropoides var.mandshuricum roots and rhizome was 57.50%,microwave radiation power of 540 W,and continuous microwave irradiation for 40 min,we can get the highest yield of essential oil was 15.80±0.55 mL/kg.Gas chromatography-mass spectrometry was used to identify the essential oils of A.heterotropoides var.mandshuricum roots and rhizomes.The results showed that methyl eugenol,3,5-dimethoxytoluene,3,4,5-trimethoxytoluene and β-pinene are the main components of essential oil.The relative amount of these four components are 40.33%,12.47%,12.74%,and 1.62%,respectively.Compared with traditional essential oil extraction methods,microwave-assisted steam distillation has the advantages of high efficiency of extraction of essential oil,low energy consumption and short extraction time.2.Microwave-assisted steam distillation followed by extraction method(MSDE)was used to extract the fat-soluble components(asarinin,sesamin)from A.heterotropoides var.mandshuricum roots and rhizome,and the asarinin and sesamin were determined by single factor experiment and Box-Behnken design(BBD)method.The optimal extraction conditions are as follows:under 90%ethanol volume fraction,19 mL/g ratio,332 W microwave radiation power and 20 min microwave irradiation time,fat-soluble components,asarinin and sesamin,were 1.81±0.27 and 0.74±0.09 mg/g,respectively.A method for the simultaneous determination of asarinin and sesamin by high performance liquid chromatography was established.The chromatographic conditions were:column:Agilent 5 TC-C18(5 μm,4.6 mm×250 mm);column temperature:28℃ detection wavelength:287 nm;injection volume 10μL;flow rate 1 mL/min;mobile phase composition and elution ratio:0.5%phosphoric acid:acetonitrile(v:v=50:50);isocratic elution.We compared the yield of asarinin and sesamin from the residue obtained by extracting the essential oil by the conventional method with residue obtained by extracting the essential oil by using a microwave-assisted steam distillation method.The results showed that the MSDE extraction process had a higher yield for extracting the fat-soluble components(asarinin and sesamin),indicating that the method of this paper is an efficient and rapid extraction method.3.The antifungal activity test of the extracted essential oils of A.heterotropoides var.mandshuricum roots and rhizomes showed that the minimum inhibitory concentration(MIC)and minimal bactericidal concentration(MBC)of the five Fusarium species were:F.avenaceum MIC=1.25 mg/mL,MBC=1.5 mg/mL;F.trichothecioides MIC=1.5 mg/mL,MBC=1.5 mg/mL;F.sporotrioides MIC=1.5 mg/mL,MBC=2.0 mg/mL;F.sambucinum MIC>2.0 mg/mL,MBC=3.0 mg/mL;F.culmorum MIC>2.0 mg/mL,MBC=2.5 mg/mL.The result showed that essential oils had obvious antibacterial activity against Fusarium spp.,inhibiting the growth and destruction of hyphae.The morphology of the filaments inhibits the production and development of spores.The use of microwave-assisted steam distillation technology can not only obtain essential oils,but also achieve efficient production of fat-soluble components(asarinin and sesamin).The optimal experimental conditions for extracting asarinin and sesamin were obtained by single factor experiment and response surface optimization experiment.Experiments prove that microwave water extraction technology is a new,efficient and resource-saving method. |
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