Extraction Technology And Bioactivity Research Of Volatile Components From Pinus Crassicorticea And Pinus Yunnanensis Franch.Var Tenuifolia

The extraction technology, chemical composition, and biological activity ofessential oils from pine needles of Pinus crassicorticea and Pinus yunnanensisFranch. var tenuifolia were investigated. The main research findings were as follow:（1） The essential oil from Pinus yunnanensis Franch. var tenuifolia was obtainedby steam distillation. The influence of solid-liquid ratio, extraction time and soakingtime on the yield of essential oil were investigated. The orthogonal test showed thatoptimum technology of extracting essential oil was follows: solid-liquid ratio was1:12, extract time was2h, and soaking time was4h.（2） Chemical components of essential oils from Pinus crassicorticea and Pinusyunnanensis Franch.var tenuifolia were analyzed by gas chromatography-massspectrometry （GC-MS） method. The major constituents of the volatile componentsfrom Pinus crassicorticea were α-pinene, β-pinene, β-caryophyllene, terpinolene and1,2,4a,5,8,8a-hexahydro-4,7-dimethyl-1-（1-methylethyl）-naphthalene. The majorconstituents of the volatile components from Pinus yunnanensis Franch. vartenuifolia were α-pinene, β-caryophyllene,1,2,4a,5,8,8a-hexahydro-4,7-dimethyl-1-（1-methylethyl）-naphthalene, β-cubebene and elixene.（3） The antibacterial activities of essential oils from needles of Pinuscrassicorticea and Pinus yunnanensis Franch. var tenuifolia against bacteria, mold,yeasts were tested using a paper disk method. The essential oils were then tested insolid medium to determine minimum inhibitory concentration （MIC） by agar dilutionmethod. The MIC of essential oil from Pinus crassicorticea against tested bacteria（Staphylococus aureus, Bacillus subtilis, Escherichia coli, Salmonella sp.） were40mL·L^-1,40mL·L^-1,40mL·L^-1,40mL·L^-1, and>80mL·L^-1,>80mL·L^-1,>80mL·L^-1,20mL·L^-1for mold （Aspergillus flavus, Aspergillus niger, Penicillum citrinum,Rhizopus sp.） and160mL·L^-1,80mL·L^-1,80mL·L^-1for yeasts （Candida Berkhout,Saccharomyces cerevisiae, Rhodotorula sp.）. The MIC of essential oil from Pinusyunnanensis Franch. var tenuifolia against tested bacteria （Staphylococus aureus,Bacillus subtilis, Escherichia coli, Salmonella sp.） were40mL·L^-1,40mL·L^-1,40mL·L^-1,40mL·L^-1, and80mL·L^-1,80mL·L^-1,80mL·L^-1,80mL·L^-1for mold（Aspergillus flavus, Aspergillus niger, Penicillum citrinum, Rhizopus sp.） and160mL·L^-1,160mL·L^-1,80mL·L^-1for yeasts （Candida Berkhout, Saccharomycescerevisiae, Rhodotorula sp.）. （4） The antioxidant activities of the essential oils from needles of Pinuscrassicorticea and Pinus yunnanensis Franch. var tenuifolia were assessed byscavenging DPPH free radical and hydroxyl free radical. The results showed that bothof the essential oils had certain scavenging activities on free radical. IC₅₀values ofscavenging DPPH free radical and hydroxyl free radical for the essential oil of Pinuscrassicorticea were7.06mg·mL^-1and0.48mg·mL^-1respectively. IC₅₀values ofscavenging DPPH free radical and hydroxyl free radical for the essential oil of Pinusyunnanensis Franch. var tenuifolia were12.16mg·mL^-1and0.38mg·mL^-1respectively.（5） The essential oils from Pinus crassicorticea and Pinus yunnanensisFranch.var tenuifolia were tested for repellency against Culex pipiensquinquefasciatus accorrding to the national standard of GB/T139179.9-2009. At thesame time, the fumigant activities were investigated by airtight fumigation in conicalflask. At the concentration of30.0%,40.0%,50.0%, the effective protection time forhuman body of essential oil from Pinus crassicorticea against Culex pipiensquinquefasciatus were30min,120min,300min. The effective protection time ofessential oil from Pinus yunnanensis Franch.var tenuifolia were30min,90min,270min. When the fumigating time was0.5h,1h,2h,4h,8h,24h, the median lethalconcentration (LC₅₀) of essential oil from Pinus crassicorticea were10.382%,2.157%,1.279%,0.669%,0.414%,0.309%. The median lethal concentration (LC₅₀)of essential oil from Pinus yunnanensis Franch.var tenuifolia was4.717%,0.970%,0.932%,0.646%,0.415%,0.216%. At the concentration of6.4%, the median lethaltime (LT₅₀) of essential oil from Pinus crassicorticea against Culex pipiensquinquefasciatus was35.167min. The median lethal time (LT₅₀) of essential oil fromPinus yunnanensis Franch.var tenuifolia was34.955min.